In 2009, the U.S. Food and Drug Administration determined that dried baobab fruit pulp was generally recognized as safe for use as an ingredient in fruit drinks up to 10 percent and fruit bars up to 15 percent. This means that 10 grams of baobab fruit is the maximum amount allowed in a 100-gram smoothie, and about 7 grams in a cereal bar.
Ten grams of baobab fruit pulp contains five grams of fiber, 12.5 percent of the Recommended Daily Intake for Vitamin C, 4 percent of the RDI for potassium, 5 percent of the RDI for iron for women, 12.5 percent of the RDI for iron for men and 2 percent of the RDI for calcium.

Baobab tree

The fruit pulp can have more Vitamin C than an orange and exceeds the calcium content of cow’s milk. It also contains carbohydrates in the form of sugars, as well as minimal protein and minimal fat.
But is there a drawback to consuming foods containing baobab fruit pulp or a baobab fruit supplement?
The fruit produced by the baobab tree — also known as the bottle tree, upside-down tree or monkey tree — has long been a staple in the diet of people living in Africa. Since 2008, there has been increasing interest for developing baobab as a raw material for consumer products.
Three food manufacturing companies now have products containing baobab fruit on the market, either in the form of supplements or as an ingredient in food bars or smoothies, and they claim the baobab fruit is the « King of Superfruits. »
So let’s look at baobab fruit. The baobab fruit is a large gourd-shaped fruit that contains a soft, powdery pulp and kidney-shaped seeds. Traditionally it was eaten as a sweet, consumed as a refreshing drink or used as an alternative to cream of tartar in recipes.
Traditional uses of the whole fruit outside of Africa are rare, as the pulp has to be dried and processed into a fine powder to be exported.
Due to its high fiber content, baobab fruit pulp may have a laxative effect. The five grams of fiber in 10 grams of fruit pulp are comparable to the amount of fiber in a single dose of psyllium taken as a laxative. The FDA does not presently regulate supplements (but it’s coming), so there is no guarantee that a capsule of dried baobab fruit pulp contains the amount of pulp or fiber indicated on the label.
So what’s the bottom line on baobab fruit? Consumer Lab identifies that its high fiber content, nutritional content and antioxidant properties may make it an attractive addition to one’s diet. But specific health claims have not been established, and consuming moderate to large amounts has a potential laxative effect.

Abaobab the baobab website

Tags: antioxidant, Baobab health benefits, Baobab pulp Fruit Powder, Health benefits

When a little-known, exotic fruit is catapulted onto the marketplace-at-large and is assigned the prefix “super,” we tend to be somewhat skeptical. After all, the produce section is now teeming with Superfruits, and the beverage aisle can barely resist collapse under the burden of bottles filled with the magical elixirs of acai, pomegranate, and mangosteen among other juices of functional distinction. No doubt then, when I mention that baobab, a fruit that is prevalent in Africa, but virtually unheard of outside of it, really is the next superfruit to look out for, this assertion will be met with rolling eyes, and plenty of them.

You see the baobab fruit’s super powers extend way beyond nutritional claims. This isn’t to say though that it isn’t purported to be a extremely healthful. It is. According to the National Geographic blog, the fruit “contains six times as much vitamin C as oranges, twice as much calcium as milk, and plenty of B vitamins, magnesium, iron, phosphorous, and antioxidants.” Ah yes antioxidants. Where would the superfruit be without its store of antioxidants? And it’s definitely safe; the FDA says so. In fact, in the fall of 2009, the FDA approved the use of baobab in foods and drinks, and the dried fruit powder (which is actually the natural state of the fruit pulp) was assigned GRAS status. This means: generally regarded as safe. In Africa, the fruit has been harvested for centuries for its medicinal qualities. Not only is it considered a general cure-all tonic, but it’s also commonly used particularly to treat fevers, malaria, gastric problems, and vitamin C deficiency among other ailments.

Let’s backtrack a little. The baobab fruit or pod basically resembles an oval gourd, or a slim downed version of a watermelon. The outside, with its woody shell and velvety green coating, admittedly offers the more attractive vista. Cut into the fruit and you’ll find flesh colored sinewy fibers, and hiding among them, white fruits that resemble misshapen marshmallows. Only these marshmallows aren’t soft and tender, but the flesh around the oil-rich seeds is dry and powdery. Ironically, while this sounds somewhat unappealing for the gastronome, it’s actually ideal for the manufacturer. There’s no need to figure out how to transform a moisture heavy, highly-degradable product into a form amenable to further processing (I’m definitely counting this as evidence of super-ness). This powdery pulp is then merely milled and then packaged and transported to Europe and the US for use in smoothies, juices, nutrition bars, baked goods and countless other foods. And since the fruit has a high pectin content (around 25%) its offers the benefit of being a natural thickening and binding agent.

As for the taste, a whole continent of people can’t be wrong. With it sweet, tangy, and pear-like flavor profile, the baobab has been consumed for hundreds of years by locals. And it’s not just the fruit that makes for tasty grub in the form of sauces and porridges and a refreshing lemonade-type beverage when combined with water and sugar, the leaves are pounded to form a kind of relish, and seeds are roasted, ground and treated like coffee. Perhaps, however, the real testament to how “super” tasty the baobab fruit could be to you and me, is the award of the Gin Trophy to a spirit made with the fruit at the recent 2011 International Wine & Sprits Competition – the most well regarded competition of its kind. Whitney Neil London Dry Gin, is distilled with nine botanicals, and the one the company is most vocal about is the baobab fruit- which it refers to as a signature African botanical. The gin uses both the citrusy fruit pulp and the mocha-like seeds which, in combination, are said to be responsible for the distinctively bold and spritely character of the drink. So unique and superlative is the final product, that producer, Johnny Neill, has been credited with re-inventing gin.

But the most useful and profound significance of the baobab tree isn’t rooted in how tasty or nutritious it is, but rather in how much it’s helping impoverished African communities. Offering a hand in this process is the charity PhytoTrade Africa. The non-profit trade association which has developed an efficient workable system by which families are able to harvest the fruit and earn an income that can pay for healthcare, education and everyday necessities. According to the National Geographic blog, “women in Malawi are harvesting the fruits for commercial use and earning enough cash to pay children’s school fees.” According to a profile on the PhytoTrade website, Esnati, a Malawian mother, was able to send all her children to school, build a house, buy a bicycle and feed her family on the income provided by picking the baobab fruit (supplemented by having to occasionally sell rice or a goat).

If you thought that was the last of the litany of reasons why the baobab fruit deserves the Superfruit title, you’d be wrong. It’s also a… super-laxative. The fruit has a very high soluble fiber content – 5 grams of every 100 grams to be precise. Seeing that the FDA allows baobab powder to be used as an ingredient in blended fruit drinks at a level of up to 10% and up to 15% in fruit cereal bars, this could result in a laxative effect, according to a recent report by ConsumerLab.com. Per 100 grams, baobab actually has more soluble fiber than psyllium, the active ingredient used in laxatives. That’s sure super something!
source: www.forbes.com
Abaobab the baobab web site

Tags: antioxidant, Baobab health benefits, Baobab pulp Fruit Powder, Health benefits

The characterisation and bioactivity determination of Adansonia digitata L.

fruit pulp, for commercial product development.
Submitted to the School of Food Science and Envi ronmental H ealth
by Orfhlaith Brady
in partial completion of the requirements for the degree of Bachelor of Science
in Nutraceuti cals for Health and Nutrition
Dublin Institute of Technology
Cathal Brugha Street
May 2011
Declaration

The characterisation and bioactivity determination of Adansonia digitata L. fruit pulp, for commercial product development.

Acknowledgements
Fi rst and foremost I would l i ke to thank my supervi sor Dr. Catherine Barry-Ryan, for her guidance and support throughout thi s research project.
I would also like to sincerel y thank Laura Massi ni and Khadi ja Khouya for their continual assistance over the course of this project. Your constant help, patience and kindness are greatl y appreciated.
I also wish to extend my gratitude to Dr. Maria Hayes and Dr. Juan Valverde of Teagasc A shtown Food Research centre, for i ntroduci ng me to the baobab fruit.
This project woul d not have been possible wi thout the generous gi ft of the baobab fruit, from M r. Pat McDonagh and M s. Rosemary Doyle of the H eal y G rou p.
Fi nal l y, I wi sh to thank my fami l y, fri ends and colleagues for their support throughout my college years.
i
Abstract
Adansonia digitata L. is a tree native to a number of African countries, known more commonl y as the baobab tree. The fruit of thi s tree has been generati ng growi ng i nterest over the past number of years, as a result of the high levels of ascorbic acid, antioxidants and other bioactive compounds that have been extracted from the fruit pulp. In this study, baobab fruit pulp was characteri sed usi ng a number of parameters, i ncluding the content of moi sture, fat, protei n, pecti n and mi neral s present in the fruit. In addition to the characteri sati on of the fruit pulp, antioxidant assays including FRA P and D PPH were carried out, in an attempt to determi ne the antioxidant capacity of baobab extracts. A nti mi crobi al and prebiotic assays were performed. Results of the characterisation showed that the fruit pulp had a high anti oxi dant capacity, in addition to consi derabl y high l evel s of ascorbic acid. It was cl ear from the characteri sati on that the commercial si gni fi cance of baobab fruit, in terms of the food i ndustry, l ay wi th these hi gh l evel s of ascorbi c aci d. C onsequentl y, product devel opment was carri ed out in order to determi ne the added benefits that baobab fruit pul p coul d confer onto a product such as a cereal bar. A cereal bar was prepared and analysed using a number of criteri a, includi ng water activity, ascorbic acid content, colour and microbial l oad. The resul ts showed that the addition of baobab fruit pulp to the cereal bar increased the ascorbic acid content by al most 8 ti mes that of the control . It was concl uded that the addition of baobab fruit pul p to the cereal bar resulted in a ri se in mi neral l evel s, most notably potassium, as wel l as a si gni fi cant i ncrease in the l evel of ascorbic acid. The application of baobab fruit pulp in the food i ndustry coul d present the opportuni ty to label a food contai ni ng baobab as a functional food.
iii
Abbreviations
90H E  90°C H ydrophi l i c Extraction
90L E  90°C L i pophi l i c Extraction
AA      A scorbi c A ci d
A ES    Atomi c E mi ssi on Spectroscopy
ANOVA Anal ysi s Of Variance
AOAC Association of Official Analytical Chemists
ASA    Acetylsalicylic Acid
CFU    Colony Forming Unit
D PPH 2,2-D i phenyl -1-Pi cryl hydrazyl
EU       European Union
FRA P Ferri c Reduci ng A nti oxi dant Power
FSA I  Food Safety A uthori ty Of Irel and
GRAS G eneral l y Recogni sed As Safe
HAT    H uman A fri can T rypanosomi asi s
H E      H ydrophi l i c Extract
IC P-OE S Inducti vel y C oupl ed Plasma O pti cal E mi ssi on Spectrometry
IFAT   International Fair Trade Association
IUC N International Union for Conservation Of Nature
LE       L i pophi l i c Extract
M C F M cfarl and Standards
ORAC Oxygen Radical Absorption Capacity
ORS    Oral Rehydration Salt
PCA    Plate C ount Agar
iv
PDA    Potato Dextrose Agar
PLC     Photochemi l u mi nescence
TEAC  Trolox Equivalent Antioxidant Capacity
RT H E            Room Temperature H ydrophi l i c Extraction
RT L E Room Temperature Li pophi l i c Extraction
TNTC Too N u merous To C ount
WHO  World Health Organisation
v
List of Tables
Table 2.1: Detai l s of dilution factors used as requi red, and the volume of extract and water requi red to formul ate the dilutions. Pg.: 27
Table 3.1: Moi sture content results: fruit pul p. Pg.: 43 Table 3.2: Ash content results: fruit pul p. Pg.: 44 Table 3.3: Fat content results: fruit pul p. Pg.: 45 Table 3.4: Protei n content resul ts: fruit pul p. Pg.: 46 Table 3.5: Pecti n content resul ts: fruit pul p. Pg.: 46 Table 3.6: M i neral content results: fruit pul p. Pg.: 47 Table 3.7: Ascorbic acid results: fruit pul p. Pg.: 49 Table 3.8: Titration results. Pg.: 50
Table 3.9: T i tratabl e aci dity results: fruit pul p. Pg.: 50 Table 3.10: Total phenol i c content. Pg.: 55
Table 3.11: Total F l avonoi d Content. Pg.: 56
Table 3.12: Total Tannin Content Pg.: 56
Table 3.13: DPPH results. Pg.: 58
Table 3.14: FRAP results. Pg.: 59
Table 3.15: Weights of cereal bars currently on the market, and an average of the weights. Pg.:68
Table 3.16: Two-tai l ed stati sti cal anal ysi s of Pai red Preference Test. Pg.: 69
Table 3.17: Moi sture content: cereal bar. Pg.: 71 Table 3.18: A sh content: cereal bar. Pg.:72
vi
Table 3.19: W ater acti vity l evel s of the cereal bars, on day 1 and day 8. Pg.:73 Table 3.20: W ater acti vi ty values and associ ated m i crobi al spoi l age. Pg.:74
Table 3.21: Analysis of variance for Aw. Pg.:75
Table 3.22: Content (mg/100g sample) of calcium, sodium, magnesium and potassium present in the cereal bars. Pg.:75
Table 3.23: Ascorbi c aci d content day 1. Pg.:77 Table 3.24: Ascorbic acid content day 8. Pg.:77 Table 3.25: Percentage decl i ne in l evel s of ascorbi c aci d over 7 days. Pg.:78
Table 3.26: A nal ysi s of Variance for AA. Pg.:79 Table 3.27: PCA plate cou nt day 1. Pg.:81
Table 3.28: PDA plate count day 1. Pg.:81 Table 3.29: PCA plate cou nt day 5. Pg.:82 Table 3.30: PDA plate cou nt day 5. Pg.:83
Table 3.31: Maximum number of CFU‟s permitted in forti fi ed bl ended foods. Pg.:85
Table 3.32: Analysis of Variance for colour. Pg.:87 Table 3.33: Nutritional information. Pg.:90
Table 3.34: Cereal bar cost estimation. Pg.:91
vii
List of Figures
Figure 1.1 Baobab fruit. Pg. 2
Figure 1.2 Chemical structure of ascorbic acid. Pg. 13 Figure 1.3 C hemi cal reacti on of FRA P assay. Pg. 14 Figure 1.4 C hemi cal structure of D PPH·. Pg. 15 Figure 1.5: Pri nci pl e of the O RA C assay. Pg. 16
Figure 2.1: Illustration of the method used to perform the dilutions of crude extracts in the mi croti ter plate for the anti mi crobi al assay. Pg. 33
Figure 2.2: Illustration of the method used to perform the dilutions of crude extracts in the mi croti ter plate for the prebi oti c assay. Pg.34
Figure 2.3: B l ended cereal bar mixture. Pg.35 Figure 2.4: Sensory analysis set-up. Pg.37 Figure 3.1: H unter L a b col our scal e. Pg.:51
Figure 3.2: Overal l col our change over three hours i ll ustrati ng the affect of each solution on the browning of appl e sl i ces. Pg.:52
Figure 3.3: Contribution of LE and H E to the total phenol i c content. Pg.:55 Figure 3.4: Contribution of LE and H E to the total fl avonoi d content. Pg.:56 Figure 3.5: Absorbance reading from 400-610 nm. Pg.:57
Figure 3.6: Contribution of LE and H E to the total tannin content. Pg.:57 Figure 3.7: Contribution of LE and H E to the total D PPH content. Pg.:58 Figure 3.8: Contribution of LE and H E to the total FRA P content. Pg.:59 Figure 3.9: A nti mi crobi al capaci ty of hydrophi l i c extract on Staphyl ococcus aureus. Pg.:60
viii
Figure 3.10: A nti mi crobi al capacity of l i pophi l i c extract on Staphylococcus aureus. Pg.:60 Figure 3.11: A nti microbial capacity of hydrophi l i c extract on Escherichia coli. Pg.:61 Figure 3.12: A nti microbial capacity of l i pophi l i c extract on Escherichia coli. Pg.:61 Figure 3.13: Prebi oti c acti vity of l i pophi l i c extract on Lactobacillus brevis. Pg.:62 Figure 3.14: Prebi oti c acti vity of l i pophi l i c extract on Lactobacillus brevis. Pg.:63 Figure 3.15: Prebi oti c acti vity of l i pophi l i c extract on Lactobacillus plantarum. Pg.:64 Figure 3.16: Prebi oti c acti vity of l i pophi l i c extract on Lactobacillus plantarum. Pg.:64
Figure 3.17: Exampl e of the effect of each of the processi ng techniques; the baked sampl e on the l eft and the col d pressed version on the ri ght. Pg.:67
Figure 3.18: Fl avoured cereal bars. Cereal bars contai ni ng several fl avours i ncl udi ng coconut, ci nnamon, cocoa and vani l la. Pg.:68
Figure 3.19: Mineral content comparison. Comparison of calcium, sodium, potassium and magnesi um contents (mg/100g sampl e) in baobab cereal bar and control . Pg.: 76
Figure 3.20: Illustration of the decl i ne in l evel s of ascorbi c aci d from day 1 to day 8. Pg.:78 Figure 3.21: L *a*b* values over seven days. Pg.:85
Figure 3.22: Col our Plot cereal bar. Pg.:85
Figure 3.23: Packaging, front cover. Pg.:87
Figure 3.24: Packaging, back cover. Pg.:88
ix
Table of Contents
Page
Declaration      i
Acknowledgements      ii
Abstract           iii
Abbreviations   iv
Chapter 1        1
Introduction     1
Background to Adansonia Digitata L .  2
1.2 Traditional Uses     3
1.3 N utri ti onal profile            4
1.4 B i oacti ve compounds associ ated with the Baobab roots and l eaves      4
1.5 Bioactive compounds associated with the Baobab fruit pul p          5
1.5.1 Anti-oxidant properties   5
1.5.2 Rehydration properties    6
1. 5.3 Anti-pyretic (anti-fever effect)    8
1.5.4 Anti-Inflammatory properties      8
1.5.5 Analgesic properties       9
5.6 Hepatoprotective properties .        10
1.6 Possible Applications of Baobab fruit pul p in the N utraceutical Industry   10
Objectives:       17
List of Ai ms:    18
Chapter 2        19
M ateri al s and M ethods        19
Section 1-Characteri sation and proximate analysis      20
2.1.1 Moisture Content            20
2.1.2 Ash content        20
2.1.3 Fat Content        20
2.1.4 Protei n Content  21
1.5 Pecti n Content      21
2.1.6 M i neral Content .          22
x
2.1.7 Ascorbic Acid Content   23
2.1.8 Acidity    24
2.1.9 A nti -browning capacity 25
2.1.10 C rude Extractions        26
2.1.11 Test for presence of carotenoids           27
2.1.12 Total Phenol i cs: Fol i n-C i ocal teu assay       28
2.1.13 Total Flavonoids           29
2.1.14 Total Tannins    29
2.1.15 DPPH   30
2.1.16 FRAP   31
2.1.17 Antimicrobial assay       32
2.1.18 Prebiotic assay  33
Section 2: Product Development          35
2.2.1 Reci pe devel opment     35
2.2.2 Addition of flavours        36
2.2.3 Prototype devel opment for sensory anal ysi s     36
2.2.4 Sensory A nal ysi s         37
2.2.5. Final batch preparation  38
2.2.6 M oi sture content           39
2.2.7 Ash Content       39
2.2.8 Water activity     39
2.2.9 M i neral content.            39
2.2.10 Shel f-l i fe eval uati on: ascorbi c aci d  40
2.2.11 M icrobial spoi lage      40
2.2.12 Shelf-life evaluation: colour       41
Chapter 3        42
Results and Discœsion  42
Section 1- C haracteri sati on and proxi mate anal ysi s            43
3.1.1 M oisture Content           43
3.1.2 Ash content        44
3.1.3 Fat Content        44
3.1.4 Protei n Content  45
xi
3. 1.5 Pecti n Content  46
3.1.6 Mi neral Content 47
3.1.7 Ascorbic Acid Content   48
3.1.8 Acidity    49
3.1.9 A nti -browning capacity 51
3.1.10 C rude Extractions        53
3.1.11 Test for presence of carotenoids           54
3.1.12 Total Phenolic Content. 54
3.1.13 Total Flavonoid Content           55
3.1.14 Total TanninsjProcyanidins       56
3.1.15 DPPH   57
3.1.16 FRAP   58
3.1.17 Antimicrobial assay       59
3.1.18 Prebiotic assay  62
Section 2: Product Development          66
3.2.1 Reci pe devel opment     66
3.2.2 Addition of flavours        68
3.2.3 Prototype devel opment for sensory anal ysi s     69
3.2.4 Sensory A nal ysi s         69
3.2.5. Final batch preparation  70
3.2.6 M oi sture content           71
3.2.7 Ash Content       72
3.2.8 Water activity     72
3.2.9 M i neral content.            75
3.2. 10 Shelf-life evaluation: ascorbic acid (AA)          76
3.2. 11 Mi crobi al content .     80
3.2.12 Shelf-life evaluation: colour       84
3.2.13 Packaging, marketing, nutri ti onal information and costi ngs of final product      86
Chapter 4        92
Conclusion       92
References       95
xi i

C hapter 1
Introduction

Background to Adansonia Digitata L.
Baobab (Adansonia Digitata L) is a tree found widely throughout Africa and known locally
in African countries as the “tree of life” due to its ability to sustain life owing to its water
holding capaci ty, as wel l as its many traditi onal medi ci nal and nutri ti onal uses (W i ckens & Lowe, 2008). Adansonia digitata is one of eight species of the Adansonia genus, and its name origi nates from the fact that the oblong leaves of the tree, often formed in groups of five, look like the fingers or digits of the human hand. It is a deciduous tree which has four growth phases and produces a fruit consisti ng of a yel lowish-white pulp which has a floury texture and numerous hard, round seeds, enclosed in a tough shell (Wickens & Lowe, 2008).

Figure 1.1: Baobab fruit. Image of the hard outer shell encasi ng the fruit (left), and the contents of the inside of the shell when opened (right), including the seed, white pulp and red fi brous pul p.
The fruit pulp of the tree has a high iron, calcium and potassium content (Yazzie et al., 1994) and is often consumed as a sweet snack and added to other foods to provide additional nutrients. Furthermore, baobab fruit pulp can contain up to ten times the vitamin C content of oranges (Vertuani et al., 2002). According to traditional folklore, all parts of the baobab tree may be used for their medicinal properties (Watt & Breyer-Brandwijk, 1962). The health benefits associated with the consumption of parts of the baobab tree may
2
be due to the presence of certain bioactive components that have been isolated from the leaves, fruit pulp and stalks and include plant sterols, triterpenes, saponins, tannins and gl ycosi des (Ramadan et al., 1993) and anti oxi dants. Studi es have been carri ed out in order to determine what bioactive compounds are present and what, if any, beneficial physiological effects these compounds may have on human health following consumption (V ertuani et al., 2002; V i mal anthan & Hudson, 2009; M i l i za, 2002; M asol a et al., 2009) .
The International Union for Conservation of Nature (I UCN) has predicted that the potential volume of baobab fruit harvested could reach 713,000 tonnes per year by 2012, almost four times the current volume currently being produced which is mostly destined for domestic use in the eight African countries investigated. The export of large quantities of baobab fruit may result in a multi-million dollar market and see a significant financial boost to thousands of poor rural producers who gather and sell the fruit to local processors (Greunwal d & Gal azi a, 2005; Hermann, 2009) . Fair trade pol i ci es put in place, such as the International Fair Trade Association (I FAT), mean that farmers and growers of baobab wi ll receive a fair price for their produce (Greunwald & Galazia, 2005). In addition to this, special harvesting and cultivation techniques are put in place which in turn minimise adverse envi ronmental impact (G reu nwal d & Gal azi a, 2005) . A 2006 report by marketing
economist Ben Bennett of Britain‟s Natural Resources Institute estimated that in southern
Africa alone, harvesting the baobab could generate more than $960 million worth of trade a year and employ over 2.6 million households (Bennett, 2006).
1.2 Traditional Uses
The baobab tree is an important food, water and shelter source in many African countries
(Venter & Venter, 1996). Baobab has a long history of traditional uses as a treatment against fevers, dysentery, and bleeding wounds, and it has a long history of nutritional and medical use in Africa (Gruenwald, 2009). The fruit of the tree, known as “monkey bread”, is a popular food source and may be consumed in a number of different ways. It is commonly sucked, chewed or boiled in water to make a drink known as Bouye. It can also be added as a supplement to mix with staple, less nutritious foods such as porridge, as an added source of calcium (Prentice et al., 1993). As wel l as bei ng a source of nourishment, all parts of the baobab tree including the fruit are traditionally used for their medicinal
3
properties. Fruit pulp can be eaten raw or soaked in water (Wickens & Lowe, 2008). Indeed, the pul p i s often consumed as a thi ckener in gruel (W i ckens & Lowe, 2008), in mi l k, as a ref reshi ng drink wi th col d water known as gu bdi (Bosch et al., 2004), as a baki ng alternative for cream of tartar, and as a rich source of calcium for pregnant women and chi l dren (Prenti ce et al., 1993). The fruit can be used for the treatment of fever, di arrhoea, dysentery, haemoptysi s and smal l pox (G reunwal d & G al azi a, 2005). The l eaf can al so be used for the treatment of inflammation, ki dney and bl adder di sease, bl ood clearing and asthma (G reunwal d & G al azi a, 2005; Van W i ck & G eri cke, 2000). The bark has been used for the treatment of fever, especially that caused by malaria (Greunwald & Galazia, 2005).
1.3 Nutritional profile
The fruit pulp of baobab has been reported to be rich in carbohydrate (76.2 %), l ow in protein (3.2 %), and fat (0.3 %). The pulp also has been found to have exceptional l y high l evel s of calcium approxi matel y, 295 mg/100 g (M agdi, 2004). Furthermore, Arnold et al. (1985), examined the nutritional composition of baobab fruit pulp (per 100g) and results docu mented were as fol l ows: water 8.7 g, energy 1290 kJ (308 kcal ), protei n 2.7 g, fat 0.2 g, carbohydrate 73.7 g, fibre 8.9 g, calcium 335 mg, magnesi um 167 mg, phosphorus 76.2 mg, i ron 2.7 mg, zinc 1.0 mg, thiamine 0.62 mg, ri bofl avi n 0.14 mg, ni aci n: 2.7 mg, ascorbic acid 209 mg. Variations in the nutritional composition of baobab fruit pulp may be due to envi ronmental factors i ncl udi ng different soi l and cl i mate conditions (M agdi, 2004).
1.4 Bioactive compounds associated with the Baobab roots and leaves
Baobab roots and l eaves have been i nvesti gated in an attempt to i denti fy the potenti al bi oacti ves associ ated wi th these parts of the plants (V ertuani et al., 2002; V i mal anthan & Hudson, 2009; M asola et al., 2009; Atawodi, 2005; Atawodi et al., 2003, A nani l et al., 2000). Certain bioactive compounds may be responsible for the treatment of certain ailments, as well as containing properties that can be beneficial to overall heal th. Examples of such bioactive compounds include tannins, phlorotannins, terpenoids, glycosides, saponins and terpenoids (Masol aetal., 2009) as well as antioxidants including both water­soluble and l i pi d-soluble components (Vertuanietal., 2002). For the purpose of this review, the bioactive compounds with antioxidant properties and potential antimicrobial activity present in the fruit pulp will be discussed in detail, asitis this portion of the plant that this thesis will focus on.

1.5 Bioactive compounds associated with the Baobab fruit pulp
1.5.1 Anti-oxidant properties
As a result of its high natural vitamin C content, baobab fruit pulp has a well documented antioxidant capability (Vertuani et al., 2002; Besco et al., 2007; Blomhoff et al., 2010, Lamien-Meda et al., 2008). It is for this reason that baobab fruit pulp is used as a milk substitute for babies in some African countries (Wickens & Lowe, 2008), in addition to the fact that vitamin C aids the absorption of iron and calcium into the body. Furthermore, it has been reported that Arabic sailors would consumed the fruit in order to prevent scurvy (Gruenwald & Galizia, 2005).
A nti oxi dants coul d hel p prevent oxi dati ve stress rel ated diseases such as cancer, agi ng, inflammation and cardio-vascular diseases as they may eliminate free radicals which contribute to these chronic diseases (Blomhoff et al., 2010; Kaur & Kapoor, 2001). It is well noted that a diet rich in fruit and vegetables, which contain an abundance of antioxidants such as vitami ns C and E as wel l as phenol ic compounds and carotenoids, can help prevent oxidative related diseases (Kaur & Kapoor, 2001). The antioxidant capacity of baobab fruit pulp was investigated using the Photochemiluminescence (PLC) assay, comparing the antioxidant properties of the fruit pulp to the antioxidant properties of several other fruits including kiwi, orange, apple and strawberry (Vertuani et al., 2002). The baobab fruit was found to have the highest content of vitamin C at 150-499 mg/100g, out of al l fruits investigated. This compared to a vitamin C content of 53 mg/100g in oranges, a well documented source of vitamin C. Baobab fruit pulp was found to have the highest water-soluble (6.96 mmol/g), lipid-soluble (4.148 mmol/g) and, therefore, total (11.11 mmol/g) antioxidant capacities of all the fruits investigated. The authors of this study, upon examination of the results, suggested that because of the level of antioxidant
capacity, baobab fruit is a “new valuable ingredient for food preparation and/or nutraceutical application in the promotion of health” (Vertuani et al., 2002).
In a more recent study, carried out by Lamien-Meda et al. (2008), the phenolic content and antioxidant activity of baobab fruit pulp using DPPH, FRAP and ABTS methods were examined. Extracts of the fruit were formulated using two different solvents, acetone 70% and methanol 70% (30). Acetone extracts were found to have significantly higher levels of
5
total phenol i c and fl avonoi d l evel s than methanol extracts. Out of a total of 14 speci es of fruit anal ysed, baobab was found to have the second and thi rd hi ghest phenol i c (4072.5 mg/100g) and fl avonoi d (42.73 mg/100g) content respecti vel y.
The high vitamin C and antioxidant content of the fruit pulp may have a role to play in the extension of shel f-l i fe for foods and beverages, as wel l as cosmeti cs. The food/beverage i ndustry coul d i ntroduce baobab fruit pul p i nto foods in order to act as a preservi ng i ngredi ent by preventi ng oxi dati on of l i pi ds in the food (A fol abi & Popool a, 2004). The addition of baobab fruit extract to cosmetics such as face creams and makeup could be a novel way of extendi ng the shel f-l i fe of both water-based and oi l -based products on account of the high l evel s of water-soluble and l i pi d-sol ubl e anti oxi dants present in the fruit (V ertuani et al., 2002). Baobab i s currentl y l i sted in Canada as a substance i s cosmeti cs and care products regulated under the Foods and Drug act (Wilkinson, 2006). Baobab fruit pulp coul d al so be used as a functional i ngredi ent in cosmeti c products, i nhi bi ti ng the agi ng of ski n due to the anti oxi dant effect of the pul p (G ruenwal d & G al i zi a, 2005).
1. 5.2 Rehydration properties
A cute di arrhoeal di seases are one of the l eadi ng causes of mortal ity in infants and young chi l dren in many devel opi ng countri es (W H O, 2006). In most cases, death i s caused by dehydrati on whi ch can be prevented by gi vi ng patients an adequate glucose-el ectrol yte solution by mouth (W H O, 2006). Solutions of oral rehydrati on sal ts are now the main treatment and are parti cul arl y useful when i ntravenous fl ui ds are in short supply, health services are basic, and there i s a shortage of ski ll ed staff (Hahn et al., 2001; A l mroth & Latham, 1995). In a study carried out by Tal-Dia et al. (1997), the efficacy of a traditional local solution made up of dried baobab fruit with water and sugar was compared to the Worl d H ealth Organisation (W H O) standard solution used to treat chi l dren with acute diarrhoea. The WHO standard solution is an Oral Rehydration Salt (ORS) solution consi sti ng of glucose, sodium, potassium and citrate di ssol ved in dri nki ng water. As a resul t of diarrhoea, the chi l dren in thi s trial had mi l d dehydrati on. In the trial, 79 chi l dren of 6 months or ol der were gi ven the standard W H O solution and 82 chi l dren were gi ven the local solution made up with dri ed baobab fruit. C hi l dren were then monitored for a peri od of 4-48 hours. Results obtai ned were based on the progression of diarrhoea and wei ght

gain. A l though resul ts obtai ned in thi s study reveal ed that the W H O solution was found to be superi or to the baobab mixture, there was no stati sti cal di fference between the two solutions in terms of durati on of di arrhoea and wei ght gain. It was also noted that the tradi ti onal local solution di d possess the advantages of bei ng an excellent nutri ent source, more economical than the WHO solution and also easily available to African cultures compared to the W H O solution. T hese added nutri ti onal properti es are an advantag e in combati ng the debi l i tati on associ ated with di arrhoea (Fopa, 1994). Two main factors attri buted to the anti di arrhoei c action of baobab are thought to i ncl ude the astringent action of the tannins causi ng an inhibition of osmoti c secreti ons in addition to the anti – i nfl ammatory action of the boabab mucilage on the intestinal mucous membrane (W i ckens & Lowe, 2008). The presence of tannins, mucilage, cellulose and ci tri c aci d present in the Baobab may also have a role to play in the effects of Baobab fruit pulp against diarrhoea (G ruenwal d & G al i zi a, 2005).
T hese rehydrati ng properti es, as wel l as the hi gh l evel s of calcium fou nd in the fruit coul d make baobab fruit pul p an ideal candidate for use in the infant formula market. As the pul p of the baobab fruit i s commonl y used to make a nouri shi ng drink, whi ch i s gi ven to babi es in some A fri can countri es as a mi l k substitute (G ruenwal d & G al i zi a, 2005), the theory of provi di ng nouri shment to babi es with thi s fruit i s not a new concept. Kenyan mothers al so feed the pul p fl our to babi es duri ng weani ng (W i ckens & Lowe, 2008). The hi gh calcium content (Prenti ce et al., 1993), anti -di arrhoea effects (Tal -Dia et al., 1997) and benefi ci al gastroi ntesti nal effects i ncl udi ng treatment of gastroenteri ti s and col i c (A bdal l a et al., 2010) are all properties of baobab fruit pul p which coul d contri bute to the success of baobab fruit pulp as an ingredient in infant nutrition. The dri ed baobab fruit pulp could be formulated i nto a powder and combi ned with water to make a dai ry-free alternative to exi sti ng infant formula products. The dri ed fruit pul p coul d al so be i ncorporated i nto an exi sti ng product in order to boost the overal l nutri ent content of that formula (Tal -Dia et al., 1997). An opportunity for a baobab fruit pul p-based mixture to become a dai ry-free alternative to mil k in products such as ice-creams, yoghurts, frozen yoghurts, desserts and other dai ry-based products i s also possible. The popul ari ty of lactose-free foods i s i ncreasi ng as a resul t of i ncreased avai l abi l i ty of these products as wel l as an i ncreasi ng awareness of lactose intolerance. Consequently, there may be room on the market for an additional calcium rich

alternative to milk such as a products made with baobab fruit pulp. It could also be a useful ingredient for the inclusion into the diet of elderly people. The powdered pulp could be easi l y added to meal s such as soups, porridge and other meal s as wel l as drinks in order to boost the calcium i ntake of thi s sector of the population.
1. 5.3 Anti -pyretic (anti-fever effect)
Sufferers of malaria in Africa, India, Sri Lanka and the West I ndies are said to consume a mash contai ni ng dried baobab bark as a febrifuge in order to treat the fever associated with this i l l ness (Wickens & Lowe, 2008) . Fruit pulp and seeds are al so wi del y used for thei r anti-pyretic properties (Wickens & Lowe, 2008; Ramadan et al., 1994). An infusion of the fruit pulp and water is consumed for its anti-pyretic qualities (Wickens & Lowe, 2008) while baobab fruit pulp has also been shown to lower elevated body temperature without affecti ng normal body temperature (Ramadan et al., 1993). In a study by Ramadan et al., 1994, Adansonia digitata was tested in vivo to determi ne the bi ol ogi cal anti-pyreti c activity of fruit pulp in rats. A quantity of 110 g of pulp was separated from the seeds and extraction of the pulp was carried out using hot distilled water in five aliquots of 1.5 L. The combined filtrates were then freeze dried to yield 80 g of dried sample. Four groups of hyperthermic rats were used in this study. The fi rst group was used as a control, the second was admi ni stered aspi ri n (ASA) as a positive control and group 3 and 4 were admi ni stered aqueous baobab extracts in doses of 400 mg/kg and 800mg/kg respectively. The temperature of each rat was then recorded hourly for 4 hours. The results show that there was a marked reduction of 1.94° C in the temperature of the mice that had been provided with an aqueous Baobab extract of 800 mg/mL, compared to 0.42° C reduction for the
control group. The authors pointed out that the “antipyretic activity of this extract resembled that normally induced by standard dose of ASA in hyperthermic rats”.
1. 5.4 Anti-Inflammatory properties
In Mali, swol l en joints are treated by rubbi ng a paste made with baobab fruit i nto the affected area causing a cure of the symptoms of this condition known as joint effusion (Wickens & Lowe, 2008; Baerts-Lehmann, 2002). Ramadan et al. (1994) found that the fruit pulp of Baobab has similar anti-inflammatory properties to phenylbutazone in rats. Phenylbutazone is used as an anti-inflammatory and analgesic drug (Encyclopaedia

B ri tanni ca, 2010). The fruit pul p was extracted as descri bed previ ousl y (Ramadan et al, 1993) and the anti -i nfl ammatory effects were tested on W i star rats. In order to carry out the experi ment, the thi ckness of the l eft paw of four groups of rats was measured in mi l l i metres. The fi rst group of rats was used as a control, the second group was admi ni stered with 15 mg/kg phenyl butazone, and group 3 and 4 were gi ven doses of Baobab fruit pul p extracts at 400 mg/kg and 800 mg/kg respectively. The effect of the extract on the paw of the rats was measured at 1, 2, 3, 4, 6, 8, 12 and 24 hours post treatment. The results showed that the baobab extract had i nhi bited oedema by up to 90 % of the efficacy of the phenyl butazone after 24 hours. The concentration of baobab fruit pul p requi red to cause these effects was cal cul ated to be an extrapol ated value of 12-18 g of fruit pul p. As the cal cul ated bi oacti ve dose i s greater than the suggested dai l y i ntake of a maximum of 10-15 g of the fruit (Wilkinson, 2006), an anti -i nflammatory effect may not be achieved at thi s amount. The authors stated that thi s anti -i nfl ammatory effect may be due to the presence of sterol s, saponi ns and tri terpenes in the extract. Furthermore, a study carri ed out by V i malanathan and Hudson (2009), demonstrated that the pul p extracts had anti – i nfl ammatory properti es whi ch the authors say coul d expl ai n some of the medi cal benefi ts attri buted to tradi ti onal pul p preparati ons.
1.5.5 Analgesic properties
The anal gesi c/pai nki ll i ng effect of baobab was al so i nvestigated by Ramadan et al. (1994). The baobab fruit pul p was extracted as descri bed earl i er. In order to carry out thi s experiment, 20 Balb/c mice were divided into four groups. The first group was left untreated and used as a control . G roup two was admi ni stered acetyl sal i cyl i c acid (ASA) at the dose of 50 mg/kg. G roup three and four were gi ven baobab aqueous extract in doses of 400 mg/kg and 800 mg/kg respecti vel y. The hot plate method was used as descri bed by Jacob and Bosvski (1961). The results were calculated by the reaction time for pain exhi bi ted by the mice, and showed a marked anal gesi c effect of 15 seconds at a 800 mg/kg dose whi ch resembl ed by 90% that of the effect i nduced by A SA duri ng the same ti me period.

1. 5.6 Hepatoprotective properties
The hepatoprotective influences of baobab fruit pulp were investigated by Al-Qarawi et al. (2003) . W hen extracts of baobab pulp were assessed for thei r influence on the liver of Wistar male albino rats, it was found that significant hepatoprotection was achieved. The fruit pulp surroundi ng the seeds was soaked in water in order to separate the pulp from the seeds. Once filtered, a mixture of water and fruit pulp remained which was subsequently freeze-dried. The white powder product was then tested in vivo for its efficacy in liver protection. The results showed that baobab fruit pulp extract had both protection and restoration effects of CCl4-inducted liver damage in the rats. The authors stated that this may have been as a result of the triterpenoids, β-sitosterol, β-amyri n palmitate, terpenoids, and ursolic acid present in the fruit. The authors also summarised that other bioactivities of Baobab fruit including analgesic, anti-inflammatory and antimicrobial activities could be factors i nfl uenci ng the hepatoprotective activity observed (Al-Qarawi et al., 2003).
1.6 Possible Applications of Baobab fruit pulp in the N utraceutical I nclustry
Nutraceuticals are defined as dietary supplements that deliver a concentrated form of a presumed bioactive agent from a food, presented in a non-food matrix, and used with the purpose of enhanci ng health in dosages that exceed those that coul d be obtai ned from normal foods (Zeisel, 1999). As the relationship between drug and food is constantly getting cl oser (Bernal et al., 2010), consumers are beginning to realise that the consumption of foods containing beneficial health effects can often be considered as a means to preventi on of certain ai l ments and chroni c di seases, thus hel pi ng to reduce the need for intervention and treatment with pharmaceutical drugs. Research into functional ingredients shows promising prospects for the use of such ingredients in food products, thereby creating added value for manufacturers and benefits for consumer health (Coppens et al., 2006).
Functional foods are those that, when consumed regularly, produce a specific beneficial health effect beyond their nutritional properties (Bernal et al., 2010). According to Espín et al. (2007), the boundary between nutraceuticals and functional foods is not always clear. The main difference between nutraceuticals and functional foods is the format in which they are consumed: nutraceuticals are consumed as capsules, pi l l s, tabl ets, etc. whi l e
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functional foods are always consumed as ordinary foods. Thus, when a phytochemical is included in a food formulation, it is considered a functional food. If the same phytochemi cal i s i ncl uded in a capsule i t i s consi dered to be a nutraceuti cal (Espín et al., 2007). Nutraceuticals and functional food products can therefore fall into the category of
“value added” foods. Added value is when the producer carries out some sort of processing
that enables them to sell the product for a premium price and so i ncrease profit margins.
In June 2008, the European Union (E.U.) authorised that baobab dried fruit pulp could be sol d in the E.U. as a novel food i ngredi ent, u nder Commission D eci si on 2008/575/EC. Baobab dried fruit pulp also achieved Generally Recognised As Safe (GRAS) status in September 2009. In June 2010, the Healy Group achieved Novel Food Approval for baobab dried fruit pulp throughout Europe and since then, has introduced baobab fruit pulp to the Irish market.
The use of baobab fruit pulp as an ingredient in foods such as cereal bars and smoothies could be a beneficial addition to the nutrition, nutraceutical, functional food and supplemental food markets. As consumers become more aware of the fact that the consumption of certain food can be directly linked to health, the presence of products on the market whi ch i ncorporate a health benefit i s i ncreasi ngly desi rabl e.
Accordi ng to the United Nations, one out of every five persons worldwide wi ll be aged 60 years or over by the year 2050, compared with current figures of one in 9. As a result, the nutraceuti cal and health food markets should continue to prosper as a result of a growing and aging population. A report by the market research firm Business Communications Company (BCC) Research Inc. states that the global market for nutraceuticals increased from $117.3 billion in 2007 to an estimated $123.9 billion in 2008. The firm predicts that thi s growth should continue to 2013, where the market should reach $176.7 billion.
It has been suggested that the use of baobab fruit pulp as an addition to cereal bars could have potential in the health food/nutraceutical market. As well as containing high levels of nutrients, baobab fruit pulp has been reported to be a good source of dietary fibre. This is in addition to having prebiotic properties, owing to the fact that the fruit pulp has been shown
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to stimulate the growth and metabol i c acti vi ty of benefi ci al organi sms such as Bifidobacteria.
Baobab pul p coul d al so have a range of possi bi l iti es in other foods and coul d be used in a vari ety of other products. T hese may i ncl ude:
- Soft drinks
- Fruit fi ll i ngs, jams, sauces, and desserts
- Snack bars, breakfast cereals
- H eal th supplements, botanical extracts
- Cosmetic products i ncl udi ng nai l and skin treatments
A l though product development i s a wel l establ i shed factor in the food i ndustry, nutraceutical and functional food product development are new and emerging areas in the i ndustry. N ot onl y does the new product have to be i nnovati ve and sought after, i t must al so have the added element of having a proven health benefit. In order for a new product to succeed, i t must have a good qual ity and wel l managed product devel opment process. The new product should offer a clear and competitive advantage and have the support of top management. New products tend to fai l for a number of reasons i ncl udi ng a shortage of important new product i deas in certain areas, poor planning, l ack of di fferenti al advantage, costliness of the product development process and lack of management enthusiasm. The use of sensory anal ysi s i s an i nval uabl e asset that food product devel opers can avai l of in order to hel p predi ct the success of a product on the market.
Sensory anal ysi s (or sensory eval uati on) i s a sci enti fi c discipline that appl i es pri nci pl es of experi mental design and statistical anal ysis to the use of human senses (sight, smell, taste, touch and heari ng) for the purposes of eval uati ng consumer products. The discipline requi res panels of human assessors, on whom the products are tested, and recordi ng the responses made by them. B y appl yi ng stati sti cal techniques to the resul ts i t i s possible to make inferences and insights about the products under test. Sensory tests supply the product devel oper wi th val uabl e stati sti cal and opi ni onated information that can hel p in the devel opment process and produce a better overal l product.
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There i s a future for novel functi onal foods whi ch can be i ncorporated i nto popul ar exi sti ng foods such a cereal bar. The fruit pulp of the baobab could potentially tap into a lucrative market, targeted at individuals who wish to fortify their diets with ready-to-eat foods that have a proven health benefit. The commercial significance of baobab fruit pulp undoubtedly lies in its high vitamin C content which provides excellent anti-oxidant properties, as well as the high calcium content of the fruit pulp, the latter of which could be of interest to individuals who may not have sufficient calcium in their diets as a result of lactose i ntol erance and other contri buti ng factors. Awareness of these two components of
the fruit could spark consumer attention in the fruit dubbed by Mintel (2008) “the new superfruit”. Furthermore, extracts of baobab fruit pulp could have potential for use on the market as a nutraceuti cal .
The vitami n C (ascorbic acid) content of the fruit pulp can be determined in the laboratory using several different techniques. Ascorbic acid (AA) is one of the most important water­soluble vitamins, naturally present in food, and widely used as food additive and antioxidant (Pénicaud et al., 2010).

Figure 1.2 Chemical structure of ascorbic acid. (Garnero & Longhi, 2010)
The ascorbic acid content of the fruit pulp will be determined in this project using a titration method (AOAC method 45.1.14) which involves the use of 2, 6-dichloroindophenol sodium salt, an oxidation-reduction indicator dye, as the titrant. This method works on the pri nci pl e that ascorbic acid reduces 2,6-di chl oroi ndophenol to col orl ess solution. At end point, excess u nreduced dye i s rose pi nk in acid solution. Ascorbi c acid i s extracted and the titrati on i s performed in the presence of H PO3–CH3COOH solution, in order to mai ntai n
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proper acidity throughout the reaction and to avoid the autoxidation of ascorbic acid at high pH (AOAC, 2006). Other methods that could be utilised for the purpose of determining the ascorbic content of the fruit pulp include liquid chromatographic methods (Rodríguez­Bernaldo de Quirôs et al., 2009), in addition to UV spectrophotometric methods (Garnero & Longhi, 2010). Use of the chromatographic method can eradicate the need for sample treatment, as the sample can be analysed by direct injection. This is a rapid form of analysi s, as the total analysi s ti me does not exceed 6 min. In a study by Rodríguez­Bernal do de Quirôs et al. (2009), thi s method showed a good repeatabi l ity and an excellent sensitivity. A UV spectrophotometric method is also suitable for the analysis of ascorbic acid, which provides fast and accurate results (Garnero & Longhi, 2010). For the purpose of this project, the ascorbic acid content of the fruit pulp will be determined using the titration method as outlined above, because of the availability of chemicals used in the experi ment.
Ascorbic acid is an example of an antioxidant, and has been attributed to the majority of the antioxidant power of the baobab fruit pulp. The antioxidant capacity of foods can also be determi ned in the l aboratory usi ng a number of anal yti cal techniques, i ncl udi ng Ferri c Reducing Antioxidant Power (FRAP) Assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity assay, Trolox Equivalent Antioxidant Capacity (TEAC) and Oxygen Radical Absorption Capacity (ORAC) assay.
The FRAP assay (Fig. 1.3) is based on the electron-transfer reaction between a ferric charge-transfer complex with the ligand TPTZ (2,4,6,-tripyridyl-s-triazine) and the antioxidant (Ar-OH) (Huang et al., 2005; Apak et al., 2007):

[Fe(II)(TPTZ)2]2+, λmax = 593 nm

Figure 1.3 – Chemi cal reaction of FRAP assay (Prior et al ., 2005) .
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D PPH • i s a stable ni trogen radical wi th a UV -Vis max absorption at 515 nm (Figure 1.4). The anti oxi dant reacti on agai nst D PPH • i s equi valent to a redox col ori metri c ti trati on. D PPH • has a deep violet col our that di sappears when reduced.

Figure 1.4 – Chemical structure of DPPH • (M olyneux, 2004)
The TEAC assay i s based on the suppression of the absorbance of radical cations A BTS by antioxidants in the test sample (Wang et al., 2004). The ABTS radical typically has a bl ui sh-green col our with maximum absorbance values at 645 nm, 734 nm and 815 nm (Re et al., 1999). W hen there are anti oxi dant compounds in the reacti on medium, they capture the free radical, whi ch i s transl ated i nto a l oss of col our and therefore a reduction in absorbance, corresponding quantitatively to the concentration of antioxidants present (Z ul ueta et al., 2009).
The ORAC method, developed initially by Cao et al. (1993), consists of measuring the decrease in the fluorescence of a protei n as a result of the loss of its conformation when it suffers oxidative damage caused by a source of peroxyl radicals (ROO_). The method measures the abi l i ty of the anti oxi dants in the sampl e to protect the protei n from oxi dative damage (Z ul ueta et al., 2009).
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Figure 1.5: Pri nci pl e of the O RA C assay (Brunswick L aboratori es, 2009)
In addition to the reported high l evel s of ascorbi c aci d and anti oxidants present in the baobab fruit pul p as di scussed previ ously, the hi gh calcium content coul d be of hi gh commercial si gni fi cance. The l evel s of thi s mi neral contai ned in the fruit pul p can be determi ned usi ng numerous anal yti cal techniques, i ncl udi ng atomi c absorption spectrometry (A A S) and i nducti vel y coupl ed plasma opti cal emi ssi on spectrometry (IC P­OES). AA S uses absorption spectrometry to assess the concentration of an anal yte in a sample. It works on the principle of the Beer-Lambert Law in which standards with a known anal yte content are used to establ i sh the relation between the measured absorbance and the anal yte concentration in the sampl e. IC P-OE S works in a si mi l ar way. H owever, thi s method i s a mul ti -el ement technique that provi des faster resul ts than A A S. This i s as a result of the fact that the standard solution can contai n many elements in the same vessel, in turn reduci ng standard preparati on ti me, gl assware, ti me requi red to i ntroduce the standard solution to the instrument.
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Objectives:
This project wi ll be di vi ded i nto two main segments, section 1 and section 2. Section 1.
This wi ll i nvol ve the compl eti on of proxi mate anal ysi s of the baobab fruit pul p, in order to i denti fy the l evel s of macronutri ents and mi neral s present in the sampl e. The macronutri ents under investigation wi ll be water content, crude protei n, crude l i pids and ash content. L evel s of pecti n, ascorbi c aci d and aci di ty wi l l al so be under investigation. In terms of mi neral composition, the sampl e wi l l be anal ysed for l evel s of magnesi um, calcium, potassium and sodium. The anti -browning and anti oxi dant capacity of the fruit pul p wi l l be assessed, as wel l as the anti mi crobi al and prebi oti c capaci ty of the fruit.
Section 2.
This section wi ll i nvolve the incorporation of the fruit pul p i nto a cereal bar. For the cereal bar, a method of col d-pressing wi l l be i nvesti gated in order to bi nd the product, in addition to a heati ng/baki ng step. The product wi l l be desi gned in order to possess the fol l owi ng traits: sui tabl e for vegetari ans and coel i acs, lactose free, l ow G I, no additives, preservati ves, col ouri ngs, added sugar, syrups, dai ry, wheat or yeast. Once the most appropri ate reci pe i s deci ded on, product devel opment wi l l take place. This wi l l i nvol ved preparation of vari ous prototypes and optimisation of these prototypes in terms of the foll owi ng:

Sensory attri butes (i ncl udi ng taste, appearance and smel l) M i crobi ol ogi cal Safety G eneral deteri orati on (i ncl udi ng ti me taken for l evel s of ascorbi c aci d to deteri orate)

Once the most promi si ng prototype i s i denti fi ed wi th the ai d of sensory anal ysi s and focus group feedback, the successful reci pe wi l l be used for the remai nder of the product devel opment process. The final product wi l l be subjected to tests such as shel f-l i fe anal ysi s, col our anal ysi s, water acti vi ty and ascorbi c aci d content. T heoreti cal packaging and l abel l i ng concepts wi l l al so be devel oped, and a bri ef marketing plan compi l ed.
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List of Aims: Fruit Pulp
Eval uate moi sture, ash, fat, protei n, pecti n and mi neral content (i ncl udi ng potassium, calcium, sodium and magnesi um).
Eval uate ascorbi c aci d content, aci di ty and anti -browni ng capacity.
C rude extractions for the purpose of testi ng for the presence of carotenoi ds and eval uati ng the total phenol i cs, f l avonoi ds and tannins.
Investi gate the antioxidant capacity by employing the DPPH and FRAP assays. Investi gate the anti mi crobi al and prebi oti c capaci ty.
Product
Product devel opment
Sensory anal ysi s
Eval uati on nutriti onal value, in terms of of moi sture, ash, mi neral content (i ncl udi ng potassium, calcium, sodium and magnesi um) and mi crobi ol ogi cal l oad.
Shel f-l i fe eval uation based on water activi ty, col our and ascorbi c aci d content over a storage period of 7 days at room temperature.
Theoreti cal packaging, marketing, nutri ti onal information and costi ng of final product.

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C hapter 2
Materials and Methods
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Section 1-Characterisation and proximate analysis
2.1.1 M oisture Content
The moi sture content of the dried fruit pulp was determined by dryi ng 4g of sample in an
oven (QualivacTM, LTE, UK) at 105 °C for 24 hours and finding the percentage moi sture l oss. Each sampl e was represented in tri pl i cate.
2.1.2 Ash content
The ash content of the fruit pulp powder was determi ned by i nci nerati ng the dried samples
from section 2. 1.1 at 550 °C for 4 hours in a furnace (Carbol ite, UK).
2.1.3 Fat Content
The fat content of baobab fruit pu l p was determined using Soxhlet extraction. In order to
carry out fat determination, a 3.00 g portion of fruit pulp was weighed and extracted with 150 ml of petroleum ether (Sigma-Aldrich, Germany) for 6 hrs on a heating mantle. Each experi ment was carri ed out in tri pl i cate. Pri or to the experi ment bei ng carri ed out, the round bottom fl asks contai ni ng anti -bu mpi ng granules were dri ed in an oven in order to prevent moi stu re presence. The glassware was al l owed to cool in a desi ccator cabinet and wei ghed (W1). 50 mL petroleum ether was added to the round bottom flask which was placed on a heating mantle in the fume hood. A thimble containing the pre-weighed fruit pulp was placed i nto the Soxhlet extractor and the extractor was connected to the round bottom flask. Approximately 100 mL additional petroleum ether was added to the Soxhlet extractor until the filter paper thimble was wet and the petroleum ether ran through the Soxhlet apparatus and into the round bottom flask. The extractor was connected to a condenser and the heating mantle was turned on. The petroleum ether was boi led for 6 hours. After 6 hours, the petrol eum ether component of the fl ask was removed. This was done by al l owi ng the level of petroleum ether to rise approximately halfway up the Soxhlet extractor. The heat was turned off under the flask in order to prevent further evaporation. The condenser was removed from the Soxhlet extractor and the petroleum ether content of the extractor was poured off into a beaker. The apparatus was reattached and the heat turned on in order to resume evaporation and so increase the level of sol vent in the extractor agai n. This process of cooling and solvent removal was repeated until only a small amount of petroleum ether remained in the flask along with the petroleum ether soluble compounds (i.e. fat). The remainder of the petroleum ether was removed by evaporation by placing the round bottom
20
fl ask in an oven and wei ghi ng the fl asks peri odi cal l y unti l a constant wei ght was achi eved (W2).
2.1.4 Protein Content
The protei n content of baobab fruit pul p was determi ned usi ng the K jel dahl method consi sti ng of three steps:
Digestion: 1.00 g of sample was mixed with 12 g of potassium sulphate (Sigma-Aldrich, G ermany), 0.5 g copper sul phate (Sigma-Aldrich, G ermany) and anti bumpi ng granules and added to a Kjeldah fl ask. 20 mL of concentrated sulphuric acid (Sigma-Aldrich, G ermany) was added to the fl ask and the mixture was fi rst heated in the digestion apparatus (Kjeldatherm, Gerhardt, UK) for 20 minutes at 250 °C. After 20 minutes, the temperature was rai sed to 400 °C for 1.5 hours, unti l a green col our appeared. 100 mL of water was added to the fl asks.
Distillation: Sodium hydroxide and heat were added to the contents of the Kjeldahl flasks in the distillation apparatus (Vapodest, Gerhardt, UK). The vapour from the flask was evaporated and condensed i nto a coni cal fl ask contai ni ng 150 mL of 4 % bori c aci d solution and i ndi cator.
Quantification: The sampl e in the coni cal fl ask was titrated agai nst an i ndi cator with 0.5 M of hydrochl ori c aci d (Sigma-Aldrich, G ermany). The ti trati on was carri ed out drop-wi se unti l the yel l ow col our, that was present pri or to the distillation step, returned. The amou nt of 0.5M HCl requi red to al l ow thi s col our change to occur was recorded and used to cal cul ate the content of protei n.
Each sampl e was represented in tri pl i cate. 2.1.5 Pectin Content
The pecti n content of baobab fruit pul p was assessed usi ng a method involving refluxing in the presence of citric acid (Sigma-Aldrich, G ermany), fol l owed by fractionation using ethanol (Sigma-Aldrich, G ermany). Each sample was represented intriplicate. 2.00 g of sampl e was homogeni sed wi th 80 mL ci tri c aci d solution and the mixture was pl aced i nto a round bottom fl ask al ong with anti bumpi ng granules. The round bottom fl ask was pl aced i nto the heati ng mantl e, connected to the reflux apparatus and heated at ± 90 °C for 45
21
minutes. A fter thi s ti me, the fl ask was cool ed under ru nni ng water and pl aced in an i ce bath for 10 minutes. The contents of the fl ask was transferred i nto four plastic tubes and pl aced in a centrifuge (Rotanta 460R, H etti ch, U K) at 2500 rpm for 8 minutes in order to separate the powdered sampl e from the l i qui d. This l i qui d layer was poured off and col l ected in a beaker, to whi ch 80 mL of 80 % ethanol was added. A gel layer began to form at the top of the l i qui d at thi s stage. More ethanol was added and the content of the beaker was swi rl ed. M i xi ng caused the gel layer to break up and disperse so it was deci ded to once agai n cool in the i ce bath and centrifuge agai n in order to col l ect the gel layer at the bottom of the plastic tubes. Once centrifugation was compl eted, the top l iqui d l ayers were poured off and di scarded. The bottom gel l ayers were col l ected in a beaker and washed wi th 70 % ethanol before bei ng pl aced in the centrifuge for a final ti me. Once the top l i qui d layer had been poured off and di scarded a second ti me, the bottom gel l ayers were col l ected and pl aced on greaseproof paper in a metal tray in order to oven dry for two hours at 45 °C . The final wei ght was recorded and used to cal cul ate the pecti n content.
2.1.6 M ineral Content
Atomi c absorption spectroscopy was used for the determi nati on of the mi neral content of baobab fruit pul p. Stock solutions were prepared (A ppendix i), from whi ch standard solutions were su bsequentl y prepared. T hese stock solutions (1000ppm) were NaCl, MgCl2, CaCl2 and K C l . F i ve concentrations of each of the stock solutions were made up (i.e. 2ppm, 4ppm, 6ppm, 8ppm and 10ppm) by usi ng a cal cul ated volume of stock solution and addi ng water to make a final volume of 100 mL for each standard.
In order to prepare the sampl e stock solutions, the ash remai ni ng from the ash from section 2.1.2 was di gested usi ng 4-5 drops concentrated hydrochl ori c aci d in each of the three cruci bl es. The di gested ash in the cruci bl es was fi l tered i nto three separate 100 mL vol umetri c fl asks, whi ch were subsequentl y made up to 100 mL wi th water (i.e. 2200 ppm). The sampl e stock solutions were di l uted 1000 ti mes, resul ti ng in a final concentration of 2.2 ppm.
The samples were introduced to an atomic absorption spectrometer (SpectrAA 110, Varian Inc., USA) and the readi ngs were recorded.
22
2.1.7 Ascorbic Acid Content
The ascorbic acid content of baobab fruit pulp was determined using AOAC method 45.1.14, titration method for ascorbic acid.
2.1.7.1 Preparation of reagents:
Extracti ng solution aceti c acid-H PO3
15 g H PO3 pellets (M erck-Chemi cal s, Germany) were pul veri sed i nto a fine powder. This was added to a 500 mL beaker contai ni ng 200 mL dei oni sed H 2O. 40 mL CH 3COOH (Sigma-Aldrich, Germany) was also added to the beaker. Once dissolved, the mixture was transferred to a vol umetri c fl ask and di l uted to 500 mL with H 2O. The vol u metri c fl ask was stored in a refrigerator u nti l required.
Ascorbic acid standard solution – 1 mg/ mL
50 mg ascorbic acid (Sigma-Aldrich, Germany) was wei ghed and di l uted to 50 mL usi ng extracting solution (A) in a volumetric flask, immediately before use.
Indophenol standard solution 2.1.7.2 Preparation of DCPI P:
50 mg 2, 6-di chl oroi ndophenol Na sal t (Fl uka, Germany) was added to a 100 mL beaker, along with 42 mg NaHCO3 (Sigma-Aldrich, Germany). 50 mL deionised water was added to the beaker. The beaker was placed in a bath sonicator (Ultrasonic, US) for 10 minutes in order to mix. The mixture was transferred to a 200 mL volumetric flask and diluted with H 2O to 200 mL. The vol u metri c fl ask was covered in tin foi l in order to block out l i ght, and placed in a refri gerator.
2.1.7.3 Determination of concentration of mg ascorbic acid / mL DCPI P solution:
A titration was carried out in order to determi ne the concentration of DCPI P as mg ascorbic aci d / mL DCPI P solution. For the purpose of thi s ti trati on, i ndophenol standard solution was pl aced i nto a burette. 5 mL extracti ng solution (A) was added to an erl enmeyer fl ask along with 2 mL ascorbic acid standard solution (B). Titration was carried out until a light but distinct rose pi nk col our remai ned for 5 seconds.
23
2.1.7.4 Deternination of other reducing substances:
A ti trati on was carri ed out in order to determi ne the possible amount of reduci ng substances other than AA (bl ank). This was done by addi ng 7 mL extracti ng sol vent to an erl enmeyer flask. To thi s, 16.5 mL water was added (the same volume of water as volume DC PIP used for ti trati on above). 2 drops i.e. 0.1 mL of i ndophenol standard solution (C) was requi red for a l i ght rose col our to appear.
2.1.7.5 Sanple Preparation:
2.00 g baobab fruit pul p was added to 30 mL extracti ng solution in a plastic tube. This was performed in tri pl i cate. The mixtures were combi ned usi ng a bench-top homogoni ser (U l tra-Turrax T25, Jani ke & K unkel, Canada) for one minute and placed in a centrifuge for 8 minutes at 2500 rpm in order to separate the pul p portion from the l i qui d portion. The l i qui d portion was poured off each sampl e and i nto label l ed vol u metri c fl asks. The volume was then brought to 50 mL wi th dei oni sed water. T i trati on was carri ed out usi ng the prepared solutions. 10 mL of each solution was titrated usi ng i ndophenol standard solution (C) unti l a rose pi nk col our appeared.
2.1.8 Acidity
The aci di ty of Baobab fruit pul p was determi ned usi ng a ti trati on method.
The sampl e of baobab fruit pul p was prepared. This was achi eved by wei ghi ng 1 g of sampl e and addi ng 20 mL of water i nto a plastic tube, in tri pl i cate. The sampl es were homogeni sed in order to mi x and then pl aced in a centrifuge for ei ght minutes at 2500 rpm in order to separate the sol i d matter from the l i qui d. The top l i qui d layer was poured off each sampl e i nto correspondi ng 25 mL vol umetri c fl asks and each sampl e was made up to 25 mL. The contents of each of the three vol umetri c fl asks were poured i nto three 100 mL beakers and 25 mL of water was added to each beaker in order to produce a final volume of 50 mL for each sampl e.

In order to carry out the ti trati on, the fi rst beaker of sampl e was pl aced on a magneti c sti rri ng plate (B i bby H B 502, U K) and connected to the probe of a pH meter (Orion 420A, Thermo, US) whi ch had previ ousl y been cal i brated. A burette contai ni ng NaOH solution was also connected to this apparatus. The initial pH of the sample was recorded. The
24
ti trati on was carri ed out drop-wi se unti l a pH value of j ust bel ow 8.1, as wel l as a pH of j ust above 8.1 was recorded. The amount of NaOH requi red to reach exactl y 8.1 coul d then be calculated.
2.1.9 Anti-browning capacity
The anti-browning capacity of baobab fruit pulp was assessed. This was achieved by compari ng the abi l i ty of baobab fruit in solution to prevent browning of appl e sl i ces, wi th other solutions known to possess anti -browni ng properti es. The solutions used in order to carry out thi s compari son were:
- Baobab (1%) solution in water
- Control (water)
- Baobab (1%) potassium phosphate buffer (PBS) – Control (PBS)
- Ascorbic Acid (AA) 0.5% solution (water) – Ascorbic Acid (AA) 0.5% solution (PBS) – Sodium M etabi sul phate (SM B) 0.1 %
1 % Baobab solution in water was prepared by homogeni si ng 5 g of baobab fruit pul p in 500 mL water in an i ce bath for 1 minute, twi ce. The solution was then fi ltered through fi lter paper and then agai n through vacuum filtration.
PBS was prepared by addi ng 10.2 g potassium phosphate (Sigma-Aldrich, G ermany) to 1350 mL water. Phosphori c aci d was added in order to bri ng the pH to 2.8 (approx. 1.87 g). The solution was then made up to 1500 mL wi th water .
SM B 0.1% solution was prepared by addi ng 0.5 g of sodium metabi sul phi te (Sigma-Aldrich, G ermany) to 500 mL water

25
1 % Baobab solution in PBS was prepared by homogeni si ng 5 g of baobab fruit pul p in 500 mL PBS in an i ce bath for 1 minute, twi ce. The solution was then fi ltered through fi lter paper and then agai n through vacuum filtration.
% AA solution in water was prepared by addi ng 2.5 g of ascorbi c aci d to 500 mL water.
0.5 % AA solution in PBS was prepared by adding 2.5 g of ascorbic acid to 500 mL PBS.
Gala (red) appl es were sl i ced and di pped i mmedi atel y for 2 minutes in a correspond i ng solution. Each solution was represented in tri pl i cate. The sl i ces were removed from the solution, patted dry wi th paper, and pl aced in petri di shes. The col our (L *, a*, b*) of each sl i ce was assessed six ti mes (three ti mes on each si de of the sl i ce) usi ng a col or measurement spectrophotometer (Col orQuest® XE, HunterLab, USA) immediately i.e. T0. The col our was then assessed at every hour for 3 hours T1, T2 and T3. The data col lected from the readi ngs was then used for the cal cul ati on of ti me-dependant col our di fference (delta E) compared to initial colour. The data was also used to determine a value for 100- L *, Chroma and Hue angle.
2.1.10 Crude Extractions 2.1.10 .1 Extraction with water
Extraction with water was used to isolate the hydrophilic components of the fruit pulp. In order to carry out extraction, two di fferent temperatures were used, room temperature (RT H E) and 90 °C (90H E). A l l extractions were carri ed out in tri pl i cate. 1.00 g of sampl e was added to 20 mL di sti ll ed water, at room temperature and at 90 °C. The mixtures were homogeni sed for 1 minute and centri fuged for 8 minutes at 2500 rpm. The top l i qui d layer was fi l tered and stored at 4 °C . The pellet remai ni ng in the centrifuge tube was extracted twi ce more. This was achi eved by homogeni zi ng the pellet agai n usi ng two portions of 10 mL water. The top l i qui d l ayers were removed, fi l tered and stored wi th the previ ousl y col l ected l i qui d layer. The volume was made up to 40 mL. The extracts were frozen unti l requi red. The pellets that remai ned were extracted usi ng acetone and methanol in order to extract the l i pophi l i c components of the sampl e.



26
2.1.10.2 Extraction with acetone:methanol
A mixture of acetone (Sigma-Aldrich, G ermany) and methanol (Sigma-Aldrich, G ermany) (70:30) was prepared. The pellets previously extracted with water were extracted using the acetone and methanol mixture (RT L E, 90L E). The extraction was carri ed out on the pellets in the same manner as above, thi s time usi ng acetone and methanol i nstead of water and only at room temperature. Once all the extracts were col lected as before, the volume was made up to 50 mL with acetone:methanol (70:30). The extracts were frozen unti l requi red.
2.1.10.3 Extract dilutions
Dilutions of the extracts were prepared as requi red usi ng the fol l owi ng table as a reference.
Table 2.1: Detai l s of dilution factors used as requi red, and the volume of extract and water requi red to formul ate the dilutions.

2.1.11 Test for presenœ of carotenoids
In order to test for the presence of carotenoi ds, the 90°C l i pophi l i c extraction (90L E) and
room temperature l i pophi l i c extraction (RTL E) were used. In a separator funnel, 50 mL petroleum ether was added to the 90LE and the sample was washed. The contents of the funnel were allowed to stand until two layers formed. The sample made up the bottom layer, and thi s was removed from the funnel and washed agai n usi ng another portion of 50 mL petroleum ether. The contents were allowed to stand once more, and the bottom layer was removed. This layer was pl aced i nto a round bottom fl ask and attached to a rotary evaporator (Rotavapor ® R-210, Buchi, Switzerland) in order to evaporate the petroleum
27
ether. The contents of the round bottom flask was re-suspended in 2 mL acetone and read in a spectrophotometer (Spectronic 1201, Milton Roy, USA) at 450 nm. This process was repeated usi ng the RTL E.
2.1.12 Total Phenolics: Folin-Ciocalteu assay
The total phenol i c content of baobab crude extracts was assessed usi ng Fol i n-C i ocal teu assay, accordi ng to the procedure adopted by Wolfe and Liu (2003), with mi nor modifications. For the purpose of thi s method, the fol l owi ng were prepared:
Bulk Sample: In a test tube, 0.5 mL deionised water was added along with 0.125 mL extract and 0.125 mL Fol i n-C i ocal teu reagent (Sigma-Aldrich, G ermany). A fter 6 minutes, 1.25 mL sodium carbonate was added. The mixture was vortexed and al l owed to remai n at room temperature for 90 minutes before addi ng 1 mL of dei oni sed water and readi ng the absorbance read in a spectrophotometer at 760 nm.
Dil uted sample (1:2): A ll of the original steps, with the substitution of di l uted extract in the place of bul k extract. The di l uted sampl e was prepared by addi ng 0.5 mL of extract in 0.5 mL of water i nto an epi ndorf tube and vortexi ng.
Blank: A ll of the original steps, with the substitution of dei oni sed water in the place of the di l uted extract.
Control: A ll of the original steps with the exception of the addition of 40 ppm gal l i c aci d in the place of the di l uted extract. 40 ppm gal l i c aci d was prepared by di l uti ng a stock solution of gal l i c (400 ppm) aci d ten ti mes i.e. addi ng 10 mL of 400 ppm gal l i c aci d to 90 mL water).
Sample-Blank: A ll of the original steps with the substitution of dei oni sed water i nstead of Fol i n-C i ocal teu reagent.
The data was compi l ed by usi ng the average absorbance readi ngs in order to formul ate a graph. The values of absorbance were compared to those of a standard curve of gal l i c aci d solutions (10, 20, 40, 50, 100 ppm) and the total phenol i c content was expressed as mg gallic acid equivalents (GAE)/100 g baobab fruit pulp sample. Each sample was represented in tri pl i cate.
28
2.1.13 Total Flavonoids
The total flavonoi d content of the baobab crude extracts was determi ned usi ng the Al umi num C hl ori de A ssay. For the purpose of thi s method, the fol l owi ng were prepared:
Bulk Sample: In a plastic tube, 1.25 mL dei oni sed water was added al ong with 0.25 mL extract and 0.075 mL sodium nitrite (5%). After 5 minutes, 0.15 mL aluminium chl ori de 10 % was added. After another 6 minutes, 0.5 mL sodium hydroxi de was added to the tube, along with 0.28 mL deionised water. The mixture was vortexed and the absorbance was read in a spectrophotometer at 510 nm.
Diluted Sample(1:2): All of the original steps, with the substitution of diluted extract in the place of bul k extract. The di l uted sampl e was prepared by addi ng 0.5 mL of extract in 0.5 mL of water i nto an epi ndorf tube and vortexi ng.
Blank: A l l of the original steps, wi th the substitution of water in the place of bul k extract.
Control: A l l of the original steps, with the substitution of 50 ppm catechi n solution in the place of bul k extract. The catechi n solution was prepared by addi ng 5 mg of catechi n to 100 mL water and mixing with a magnetic stirrer until dissolved.
Sample-Blank: A ll of the original steps with the substitution of dei oni sed water i nstead of sodium nitrate.
The absorbance was measured i mmediately at 510 nm and compared to a standard curve of catechin solutions (10, 20, 40, 50, 100 ppm). The flavonoid content was expressed as mg catechi n equival ents (C E)/100 g baobab fruit pul p. Each sampl e was represented in tri pl i cate.
2.1.14 Total Tannins
The total tannin content of the baobab crude extracts was determined by the Acid-Butanol A ssay for the identification of pro-antocyani ns/condensed tannins. For the purpose of thi s method, the fol l owi ng was prepared:
Bulk Sample: In a test tube, 1.5 mL acid butanol (95:5, 1-butanol: concentrated HCl, v/v)
was added to 250 mL sampl e. 50 µ L i ron reagent was added as a catal yst, and the tubes
were vortexed. The tubes were placed in a water bath at ± 90°C for 20 mi n. After cool i ng,
29
the resul ti ng mixture was washed with 500 µ L di sti l l ed water, vortexed and then centri fuged at 8,000 rpm for 10 mi ns.
Each sampl e was repl i cated in tri pl i cate. The resulti ng pi nk supernatant was recovered and the absorbance read at 550 nm in a spectrophotometer, usi ng a quartz cuvette. M ol ecul ar confirmation of the anthocyani di n pigment was performed in a spectrophotometer (Agi l ent 8453, U SA) by a UV -Vis scan of the absorption spectru m, between 200 and 600 nm. The solvent 1-butanol was used to blank the spectrophotometer. It was noticed that the pink anthocyani di n pigment thus obtai ned had a maximum absorbance at 549 nm, confi rmi ng the presence of the cyani di n aglycone. The total tannin content was expressed as mg cyani di n equi val ent/g sampl e.
2.1.15 DPPH
The assay for testi ng the radical scavengi ng activiti es of crude baobab extracts was devel oped usi ng a modi fi ed a procedure by M akri s et al. (2007), with reference to the protocol of Truong et al. (2007). The anti-radical capacity of the extracts was assessed usi ng D PPH (2,2-di phenyl -1-pi cryl hydrazyl ). B ul k D PPH solution was prepared by addi ng 15.8 mg D PPH to 100 mL of 96 % ethanol . The bul k solution was stored in the fri dge unti l requi red. The bul k solution was di l uted 1:5 with ethanol i mmediatel y before use. For the purpose of thi s assay, six dilutions (1:2, 1:5, 1:10, 1:20, 1;50 and 1:100) of the extracts were made up, in dupl i cate, i nto 1 mL eppendorf tubes.
The fol l owi ng were al so prepared:
Bulk Sample: In an eppendorf tube, 100 uL of bul k sampl e was added to 900 uL di l uted D PPH. The mixture was l eft for 30 minutes at room temperature, in the dark, and then read in a spectrophotometer at 515 nm.
Diluted Sample: The same procedure as above was used, with the exception of the use of each of the di l uted extracts in place of the bul k sampl e.
Blank: 1 mL dei oni sed water.
Control: The same procedure as above was used, with the exception of the use of water i nstead of sampl e.
30
Each sample was represented in tri pli cate. The inhibition values were compared to those of a standard cali brati on cu rve of L-ascorbi c acid solutions. For the li pophi l i c extractions, the ascorbic acid solution was prepared using methanol in the fol lowing concentrations: 10, 20, 40, 50 and 80 ppm. For the hydrophi l i c extractions, the ascorbi c aci d solution was prepared using water in the fol l owi ng concentrations: 10, 20, 40, 50, 100 ppm. The values were expressed as mg ascorbic acid equivalents (AAE)/g of fruit pulp.
2.1.16 FRAP
The antioxidant capacity of crude baobab extracts was evaluated using a modified FRAP assay procedure based on the method by Maksimović et al. (2005).
For the purpose of thi s method, the foll owi ng were prepared:
Bulk Sample: 0.1 mL bu l k sampl e was added to an eppendorf tube.
Blank: Al l of the original steps, with the substitution of water in the place of extract.
Dil uted Samples: Several dilutions of the extracts were prepared as previ ousl y outl i ned. 0.1 mL of each of these dilutions was added to i ndi vi dual eppendorf tubes.
Sample-Blank: All of the original steps with the substitution of dei oni sed water instead of FRA P reagent.
Each sampl e was represented in tri pl i cate. A freshl y prepared FRAP reagent (100 ml acetate buffer, 300 mM, pH 3.6 + 10 ml 10 mM TPTZ (2,4,6-tripyridyl-5-triazine) in 40 mM HCl + 10 ml 20 mM FeCl3·6 H2O) was heated in water bath at 37°C for 5 min before bei ng transferred (0.9 mL) i nto the above eppendorf tubes. The tubes were pl aced in a water bath at 37°C for 40 minutes; the absorbance was read in a spectrophotometer at 595 nm. Antioxidant activities were calculated from a calibration curve of L-ascorbic acid standard solutions under the same experi mental conditions. For the lipophilic extractions, the ascorbi c aci d solution was prepared usi ng methanol in the fol l owi ng concentrations: 5, 10, 20 and 40 ppm. For the hydrophi li c extractions, the ascorbi c aci d solution was prepared using water in the following concentrations: 10, 20, 40, 50 ppm. Antioxidant activity was expressed as mg ascorbic acid equivalents (AAE) per gram of fruit pulp.
31
2.1.17 Antimicrobial assay
The antimicrobial capacity of the baobab crude extracts was assessed using a microtiter
plate reader (E L 808, B i otek, USA) and two bacteri al strai ns, Staphylococcus aureus, a Gram positive bacteri a, and Escherichia coli, a Gram negati ve bacteri a. In order to carry out thi s method, the procedure was fi rst i nputted i nto the instrument. This i ncl uded the fol l owi ng speci fi cati ons:

The cultures were grown overnight in l i qui d broth. Each culture was added dropwi se i nto a saline solution unti l 0.5 M C F was recorded (i.e. 106 C F U ). 0.1 mL of thi s prepared saline solution was added to 9.9 mL broth. 200 µ L of the S. aureus broth solution was added to wel l A 1 and wel l D 1. 200 µ L of the E. coli broth solution was added to wel l B 1 and wel l E 1. 100 µ L of broth contai ni ng S. aureus was l oaded i nto wel l s A 3-8 and D 3-8. 100 µ L of broth contai ni ng E. coli was l oaded i nto wel l s B 3-8 and E 3-8.
The l i pophi l i c extract was prepared by evaporati ng off the acetone: methanol component of the extract and resuspendi ng the dri ed matter in the equi valent volume of water. This resul ted in both the hydrophi l i c and l i pophi l i c extracts bei ng suspended in water. 200 µ L of the l i pophi l i c extract was added to wel l A2 and and B2 and mi xed. 100 µ L of the hydrophi l i c extract was added to wel l D 2 and E 2 and mi xed. 200 µ L of the contents of each of these wel l s was added to the wel l to the ri ght, as i l l ustrated bel ow in figure 2.1 This dilution was repeated unti l wel l 8 was reached.
32
100 µL 100 µL 100 µL 100 µL 100 µL
A
B
C
D
E
F
G
H
1          2 3 4 5 6 7 8 9 10 11 12
Li pophi l i c Extract
N ot used Hydrophi l i c Extract
Figure 2.1: Illustration of the method used to perform the dilutions of crude extracts in the mi crotiter plate for the anti mi crobi al assay.
Where  = Staphylococcus aureus,        = Escherichia coli,        = Bul k,           = 10-1,            =
10-2,   = 10-3,            = 10-4,            = 10-5,            = 10-6
The plate was then placed into the apparatus for 24 hours. The resulting data was used to create graphs that i l l ustrated the l evel of mi crobi al inhibition created by the vari ous concentrations of baobab extract.
2.1.18 Prebiotic assay
The prebi oti c capacity of the baobab crude extracts was assessed usi ng the method
di scussed in section 2.1.18, wi th some adj ustments. The assay procedure was carri ed out in the same way as before, usi ng the same dilutions. A djustments i ncl ude the use of Lactobacillus brevis and Lactobacillus plantarum bacterial strai ns, as well as a change in terperature from 37 °C to 30 °C.



33
100 µL 100 µL 100 µL 100 µL 100 µL

A
B
C
D
E
F
G
H
1          2 3 4 5 6 7 8 9 10 11 12
Li pophi l i c Extract
N ot used Hydrophi l i c Extract
Figure 2.2: Prebi oti c mi crotiter plate. Illustration of the method used to perform the dilutions of crude extracts in the mi croti ter plate for the prebi oti c assay.
Where  = Lactobacillus brevis, = Lactobacillus plantarum,       = Bul k,           = 10-1,
= 10-2,            = 10-3,            = 10-4,            = 10-5,            = 10-6



34
Section 2: Product Development
2.2.1 Recipe development
The cereal bar reci pe was desi gned to contai n the fol lowi ng ratio of i ngredi ents:

The i ngredi ents were pl aced in a food processor together and bl ended unti l combi ned (Figure 2.3). It was deci ded that each bar woul d wei ght 42g. Two processi ng techniques were i nvesti gated, col d pressing and baki ng. C ol d pressing i nvol ved the application of pressure to the prepared cereal bar mixture in order to bi nd the product and produce a a uni form, flat shape whi ch coul d consequentl y be cut to si ze. The use of col d pressing el i mi nated the need for the application of heat, reduci ng nutri ti onal l oss. B aki ng was al so consi dered, as an alternative to col d pressing. The baki ng step i nvol ved moul di ng the product mixture i nto baki ng tins and pl aci ng in an oven at 140 °C for 15 minutes. A focus group was uti l i sed in order to determi ne whi ch method produced more desi rabl e properti es in the final product.

Figure 2.3: Blended cereal bar mixture.
35
2.2.2 Addition of flavours
Several di fferent fl avours were added to the basic reci pe as outl i ned previ ousl y in section
2.2.1. T hese fl avours i ncl uded the fol l owi ng:

Vanilla (1/4 spatula per 100 g) Ci nnamon (1/4 spatul a per 100 g) Cocoa powder (1 teaspoon per 100 g) Coconut (1 tabl espoon per 100 g)

A batch of each of the fl avour combi nati ons was prepared. A focus group was uti l i sed in order to determi ne general appeal of each of these fl avours.
2.2.3 Prototype development for sensory analysis
Two sets of sampl es were prepared for the purpose of sensory eval uati on, set 1 and set 2. 2.2.3.1 Set 1
Set 1 was prepared in order to be uti l i sed in a Ranki ng Test for P reference. This set i nvol ved the preparati on of four batches of the cereal bar sui tabl e for sensory anal ysi s. For each batch, enough mixture for approxi matel y 7 bars (300 g) was prepared usi ng the ratio provi ded above in section 2.2.1, with the exception of the exclusion of baobab from the formulation. The flavours were added in the amounts detailed in section 2.2.2. Batch 1 contai ned ci nnamon, batch 2 contai ned cocoa, batch 3 contai ned coconut and batch 4 contai ned no added fl avour.
2.2.3.2 Set 2
Set 2 was prepared in order to be uti l i sed in a Pai red Preference Test. This set i nvol ved the preparati on of two batches of the cereal bar. E nough mixture for approxi matel y 7 bars (300 g) was prepared using the ratio provided above in section 2.2.1, with the exception of the exclusion of baobab from the formulation. The di fference between the two batches was in terms of appearance. Batch 1 contained added oat flakes on top of the plain bar recipe, whi ch was achi eved by spri nkl i ng oats on the top surface of the cereal bars before baki ng. Batch 2 did not have any added oat flakes present.
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2.2.4 Sensmry Analysis
Two sensory tests were carri ed out usi ng the sampl es prepared in section 2.2.3. The sensory
tests consi sted of a simple test (Pai red Preference Test), in addition to a more compl ex test (Ranki ng Test for Preference).
2.2.4.1 Test Preparation:
Test sheets were prepared for each test (A ppendi x i i ). Sensory plates were prepared by di vi di ng the area on the plate i nto quarters wi th a marker and wri ti ng coded numbers on these quarters of the plate, which corresponded to numbers on the sensory sheets (A ppendix iii). The bars in each set were di vi ded usi ng a kni fe.

Figure 2.4: Sensory analysi s set-up. 2.2.4.2 Test Procedure:
The coded numbers outlined in section 2.2.4.1 were assigned to the various samples. The coded numbers and correspondi ng sampl es were recorded by the tester as i ll ustrated bel ow.
Pai red Preference Test Codes:
42 + 45 = oats
43 + 44 = no oats
37
Ranki ng Test for Preference Codes: Coconut = 248
Cocoa = 912
Cinnamon = 319
Plain = 102
Each sampl e was pl aced on the plates accordi ng to the coded system. The plates contai ni ng the ranki ng test for preference sampl es were placed on the tables of the sensory booths to begi n with. Judges were seated at sensory booths. The i ngredi ents contai ned in the sampl es were detai l ed, and j udges were i nvi ted to i nform the tester of any allergies or other reason why they may not take part. The j udges were gi ven the fol l owi ng verbal instructions: Pl ease rate the cereal bars in order of your preference, wi th 1 bei ng the most preferred sampl e, and 4 bei ng the l east preferred. Pl ease rate the sampl es accordi ng to the fol l owi ng cri teri a, and i t the fol l owi ng order:
1st: A ppearance 2nd: A roma
3rd: Taste
For the pai red preference test, the plates contai ni ng the pai red preference test sampl es were placed on the tables of the sensory booths. Judges were given the fol l owi ng verbal instructions: Eval uate each set of cereal bars accordi ng to appearance. Pl ease select the cereal bar in each set that you prefer vi sual l y.
2.2.5. Final batch preparation
As the results of the sensory tests stated that cinnamon was the most preferred flavour, it was deci ded that the final product woul d contai n ci nnamon as the flavouri ng. This resul ted in the final reci pe contai ni ng the fol l owi ng i ngredi ents:
Dates Oats

38

M i xed N uts Raisins Apple Juice Baobab fruit pulp Ci nnamon

Three batches, consi sti ng of six bars each, and contai ni ng the above i ngredi ents were prepared, using the ratios previously discussed in section 2.2.1 and 2.2.2. A control batch, consisting of six bars, containing no baobab fruit pulp was also prepared. Each bar was vacuum packed separately and stored in a cool, dry, dark place until required for testing.
2.2.6 M oisture content
The moi sture content of the cereal bars was determined as before by dryi ng 1.00 g of
sample in an oven at 105 °C for 24 hours and finding the average moisture loss of the sample. Each batch was represented once.
2.2.7 Ash Content
The ash content of the cereal bar was determined as before by incinerating the dried
samples from section 2.2.6 at 550 °C for 4 hours in a furnace.
2.2.8 Water activity
Water activity of the cereal bars was measured using a water activity meter (Aqualab,
USA). The instrument was calibrated following the user‟s manual. The samples were then pressed i nto separate di shes and pl aced i nto the instrument. The A w readi ngs were recorded at day 1 and day 8.
2.2.9 M ineral content
The mineral content of the cereal bar was determined using inductively coupled plasma
(ICP) atomic emission spectroscopy (AES), (Liberty 150, Agilent, USA). The ash samples from section 2.2.7 were used for this experiment. 5-6 drops of concentrated hydrochloric acid was added to 10 mg of ashed sample to dissolve the particles. This mixture was filtered through filter paper (Whatman no. 1) into 100 mL volumetric flasks. Deionised water was then used to bring the contents of the volumetric flasks up to 100 mL. This mixture was considered to represent 100 ppm.
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2.2.10 Shelf-life evaluation: ascorbic acid
The ascorbic acid content of the cereal bar was measured on day 1 and agai n on day 8 usi ng the ti trati on method as outl i ned in section 2.1.7, wi th sl i ght modifications. The reagents were prepared in the same way as before and the ti trati ons to determi ne the mg ascorbi c aci d / mL D C PIP solution, as wel l as the reduci ng substances, were al so carri ed out in the same manner. The sample was prepared in a different manner. 2.00 g of each batch of cereal bar was wei ghed and added to 20 mL extracti ng solution in a stomacher bag. The mixtures were pl aced in a stomacher (Stomacher 400, Seward, U K ) for 4 minutes at the medium setti ng, fol l owed by centri fugi ng for 10 minutes at 10,000 rpm in order to separate the sol i d portion from the l i qui d portion. The l iquid portion was fi l tered through fi l ter paper under sucti on and col lected in plastic tubes. The volume was then brought to 25 mL wi th dei oni sed water. T i trati ons were carri ed out usi ng these extracted sampl es. 10 mL of each sample was titrated usi ng i ndophenol standard solution (C) unti l a rose pi nk col our appeared. Each sample was represented in tri pl i cate.
2.2.11 M icrobial spoilage
The presence of bacteria, yeasts and moulds in the cereal bars was investigated. Non-selecti ve media was used in order to detect and enu merate the growth of mi croorgani sms in the cereal bar.
2.2.11.1 Material preparation:
Plate Count Agar (PCA): 3 L PDA (Fl uka, Germany) was prepared by addi ng 17.5g powdered agar i nto each of three 1 L Duran ® bottl es and toppi ng each bottl e up wi th 1 L dei oni sed water. The mixture was shaken to mi x.
Potato Dextrose Agar (PDA): 3 L PCA (Scharlau, Spain) was prepared by adding 39g powdered agar i nto each of three 1 L Duran ® bottl es and toppi ng each bottl e up wi th 1 L dei oni sed water. The mixture was shaken to mi x.
Tryptone Phosphate Water (TPW): 1 L TPW (Scharlau, Spain) was prepared by addi ng 25.5 g tryptone phosphate powder to 1 L dei oni sed water in a 1 L Duran ® bottl e with a di spensi ng pump. 9 mL of the mixture was di spensed i nto 32 test tubes. 20 mL of the mixture was also added to an additional 4 test tubes. The test tubes were subsequently capped.



40
All mixtures were placed in the autoclave (Tomy SS-325, Japan) at 121 °C for 15 minutes
to heat and steri li se. After steri li sati on and cooli ng, the agar was di stri buted i nto petri- di shes.
2.2.11.2 Sarrple preparation:
1 g of each cereal bar sampl e was wei ghed and added to 20 mL of peptone in a stomacher
bag. The mixture was agitated at “medium” for 4 minutes. 5 mL of the liquid portion of the
contents of each stomacher bag was added to empty test tubes. 1 mL of thi s was added to a test tube containing 9 mL peptone (10-1). This process was continued until 8 dilutions exi sted, i.e. u p to 10-8. 100 µL of each of these dilutions was added to the plates wi th a pipette and spread usi ng a hockey stick onto a plate containing each type agar. This was repeated in tri pl i cate. The plates contai ni ng PCA were i ncubated at 37 °C for 24 hours, and the growth recorded. The plates containing PDA were i ncubated at 30 °C for 48 hours, and the growth recorded.
2.2.12 Shelf-life evaluation: colour
The colour of the cereal bar was measured on day 1 and agai n on day 8 usi ng the
col ori meter as outl i ned in section 2.1.9.
41
C hapter 3
Results and Discussion
42
Section 1- Characterisation and proximate analysis
3.1.1 M oisture Content
The moisture content/humidity of a food is an important parameter to analyse for the
purpose of determi ni ng how the food wi l l behave duri ng processi ng and storage. It i s al so a contributing factor to the food physical characteristics of the food, such as texture, taste and appearance (Nielsen, 2010). The moisture content is a key parameter for evaluating and control l i ng the ri sk of deteri orati on of the food over storage, as a resu l t of mi crobi al spoi l age. This is because the water content of a food influences the propensity of mi croorgani sms to grow in foods (Kornacki, 2010).
The percentage moisture content of baobab fruit pulp was calculated using the following equation: Moisture content (%) = (W1 – W2) / W1 x 100
Where: W1 is weight of the initial sample, W2 is weight of the sampl e dri ed at 105 °C Table 3.1: Moisture content results: fruit pulp.

Average moi sture content: (8.75 + 9.5 + 10) / 3 = 9.42 % ± 0.63
From the results above, it can be seen that the average moisture content of the fruit pulp was fou nd to be 9.42 % ± 0.63. This moisture content is si mi l ar to that reported by M agdi (2004), who reported the moisture content of the fruit pulp to be 10.4 % ± 0.4. The difference between these results is less than 10 %. The discrepancy in the results found may be as a result of disparity of the source of the samples.
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3.1.2 Ash content
A shing invol ves incinerating the organic compounds found in the sample, such as proteins

and carbohydrates, resulting in only the mineral content of the sample remaining. Dry ashi ng was employed for the purpose of determinati on of the mi neral content of the sample.

Dry ashi ng i nvol ves the use of a furnace at temperatures up to 500-600 °C, in whi ch the sampl es are pl aced. This method vaporises water and volatiles, whi l e organi c substances are burned in the presence of oxygen in the air (N i el sen, 2010).
The ash content of baobab fruit pul p was expressed as: A sh % = (W1 / W2) x 100 Where W1 = weight of ash and W2 = initial weight of dried sample at 105 °C. Table 3.2: Ash content results: fruit pul p.

Average ash content: (6.03 + 6.08 + 6.11)/ 3 = 6.07 % ± 0.04
From the results above, it can be seen that the ash content of the fruit pul p was found to be 6.07 % ± 0.04. This figure obtai ned di ffers to the values found by N our et al. (1980) of 5.3 % and M agdi (2004) of 4.5 %, but i s cl oser to the figures found in a report commi ssi oned by PhytoTrade Afri ca (2006), detai l i ng ash content values of an average of 5.9 %. The variation in resul ts may once agai n be attri butabl e to the variation of batches and sam pl es used.
3.1.3 Fat Content
The determi nati on of the content of fat in food i s si gni fi cant for the purpose of l abel l i ng when a food i s i ncorporated i nto a food product. A food manufacturer i s l egal l y obl i ged to provi de nutri ti onal information of a food product on the label, i ncl udi ng detai l s of the fat content, under Directive 90/496, Regulation 1924/2006 (FASI, 2010). The fat content of a food i s al so important as consumers become more health consci ous and aware of
44
importance of reduci ng the fat i ntake in the di et. L ow-fat foods and food i ngredi ents can play a rol e in cateri ng for consu mers wi shi ng to reduce thei r overal l fat i ntake.
Table 3.3: Fat content results: fruit pulp.

The result obtai ned from fl ask 1 was di scarded due to error duri ng extraction process. Average fat content: (3.39+3.12)/2 = 3.26 g ± 0.19
The fat content of the baobab fruit pulp was found to be 3.26% ± 0.19. This was consi derabl y hi gher than the resul ts detai l ed by M agdi (2004) of 0.30% and of N our et al. (1980) of 0.2 %. A possible expl anati on of thi s may be that some of the seed content of the fruit may have been present in the batch of fruit pul p used for the purpose of thi s anal ysi s. The fat content of baobab seed has been reported to be 12.2 % (M agdi, 2004).
3.1.4 Protei n Content
Protei n anal ysi s has nutri ti onal, heal th, l egal, and economi c implications for the food i ndustry. As di scussed previ ousl y, the nutri ti onal information of foods, i ncl udi ng protei n content, i s a l egal l y requi red aspect of food l abel l i ng. It i s important for consu mers to be aware of the content of protei n in the foods they eat, in an effort to ensure correct nutrient i ntake and to mai ntai n a healthy, bal anced di et. In terms of the economi c impact of protei n, it has been reported that the market value for major food commoditi es i s partial l y determi ned by the content of protei n present in the food (Owusu-A penten, 2002). The Kj el dahl method of protei n anal ysi s consi sts of three main steps; 1) digestion of the sampl e in sul furi c aci d wi th a catal yst, whi ch resul ts in conversion of ni trogen to ammoni a; 2) distillation of the ammoni a i nto a trappi ng solution and 3) quantification of the ammoni a by ti trati on wi th a standard solution.
45
Table 3.4: Protein content results: fruit pulp.

Average protei n content: (2.62 + 2.18 + 2.62)/3 = 2.47 % ±0.25
The results of the analysis determined a protein content of baobab fruit pulp of 2.47 % ±0.25. This result was in li ne with the fi ndi ngs of Nour et al. (1980) and M agdi (2004), who reported levels of 2.6 % and 3.2 % respectively.
3.1.5 Pectin Content
The pectin content was cal cul ated usi ng the fol l owi ng equati on: Y i el d wei ght / initial
sample weight x 100.
Table 3.5: Pectin content results: fruit pulp.

Average % al cohol insoluble sol i ds referred to as pecti n: (51.2+52.9+48.5)/3= 50.9 % ±2.2
The pectin content of the fruit pulp was found to be 50.9 % ±2.2. This figure corresponds closely to the figures found by Nour et al., (1980) of 56 % pecti n. The authors of this study
noted that the pectin of baobab pulp has “a low degree of esterification and low a low intrinsic viscosity”. These properties, the authors stated, would result in poor gelling properties of the fruit pulp, meaning the pectin contai ned in baobab fruit pulp is of lower quality to commercial apple pectin (Nour et al., 1980). As a result, baobab fruit pulp would
46
not be useful as a setting agent. However, it has been found during the process of this project that the fruit pulp has excellent thickening properties which could theoretically be applied to the food industry for the purpose of thi ckening and bulking for smoothies and fruit j ui ces, in addition to i ts application to mousse-type desserts.
3.1.6 M ineral Content
The atomic absorption spectrometer was used to produce absorbance readings, which were in turn used to calculate calibration curves for calcium, magnesium and potassium (Appendix iv). The calibration curve for sodium was not utilised as a result of the fact that the R2 value was less than 0.90, and therefore would not provide accurate results once extrapol ated. The remai ni ng cal i brati on curves were extrapol ated to determi ne the concentration of mi neral s present in each of the baobab sampl es. The results of the mi neral as seen in table 3.6, and are expressed in terms of mg mi neral/100 g baobab sampl e.
Table 3.6: Mineral content results: fruit pulp.

The calcium content of 344.23mg was found to be present in 100 g sample. This compares to 295 mg/100 g found by Magdi et al. (2004), 655 mg / 100 g by Nour et al. (1980), 115.6 mg/100 g by Kalenga Saka et al. (1994) and 341mg/ 100 g by Glew et al. (1997). If a range of the calcium content was to be compiled, using the results published by these authors, it would range from 115mg/ 100 g — 655 mg/ 100g fruit pulp. It can be seen that this range is wide, and a significant amount of variation between batches can occur. The result of 344 mg calcium / 100 g sample, as found in thi s proj ect, fits i nto thi s range nonethel ess.
47
The level of potassium in the fruit pulp was found to be 1578.51 mg/100 g sample. This agai n fit i nto the range of between 1240 mg/ 100g (M agdi , 2004) and 2834 mg/ 100g sample (Kalenga et al., 1994) as reported in the literature.
Magnesium levels in thi s sample of baobab fruit pulp were calculated to be 417.88mg/100 g sample. Kal enga Saka et al. (1994) fou nd l evel s of thi s mi neral to be al most hal f of thi s l evel , at 209 mg/100 g sampl e whi le M agdi (2004) reported l evel s as l ow as 90 mg/100 g sample.
Although sodium was not calculated at this time as a result of the fact that the calibration curve, generated from the data provided by the instrument readings, presented an R2 value of 0.90, which was not accurate enough to be extrapol ated. H owever, the content of sodium of the fruit pulp is unlikely to exceed the maximum content found in literature namely 27.9 mg/100 g sampl e (M agdi , 2004) . Other reported l evel s of sodium i ncl ude 5.5 mg/100 g (Glew et al., 1997) and 18.8 mg/100 g (Kalenga Saka et al., 1994). Using these figures as a
guide, baobab fruit pulp could be classified as a “low sodium” product, in accordance with
sti pulated nutrition and health claims (FSAI, 2010), because the levels of sodium are below 0.12 g (120 mg) sodium per 100 g.
3.1.7 Ascorbic Acid Content
Ascorbic acid (AA) is one of the most commonly reported components of baobab fruit pulp, as it had been claimed that the fruit has more ascorbic acid than oranges, which are renowned for hi gh l evel s of ascorbic acid. Ascorbi c acid i s a powerful antioxi dant, and i s often used in the food i ndustry as an additive to extend the shel f-l i fe by hel pi ng to prevent oxi dati on in the food over ti me. Ascorbi c acid i s l i ght and heat sensitive, so care was taken throughout thi s experi ment to reduce the amount of exposure the fruit pulp had to these two factors, which in turn could degrade the ascorbic acid present in the fruit and product figures which did not reflect the content of ascorbic acid in the fruit pulp. Light degradation was reduced by pl aci ng the sampl es in the dark when not in use. The sampl es w ere al so extracted in frosted plastic tubes, so as to li mit the exposure of the powder to light. Heat degradati on was reduced by keepi ng the sampl es in ice whenever possible, parti cu l arly during the extraction process, when the friction caused by the homogeniser may have raised
48
the temperature of the sampl es. During centrifugation, a temperature of 4 °C was sel ected in order to mai ntai n a chi ll ed envi ronment around the extracted fruit pul p.
The content of ascorbic acid in the fruit pulp was cal cul ated usi ng the fol l owi ng formula:
(V1-V2) x conc. DCPIP (mg/mL) x final volume ÷ vol. of sample ÷ initial weight x 100. Where V1 is the volume of DCPI P required to cause colour change, V2 is volume of blank (calculated to be 0.1 mL), conc. DCPI P (calculated to be 0.121 mg AA/mL solution).
Table 3.7:Ascorbic acid results: fruit pulp.

An average of these figures was compi led in order to fi nd the average AA content: (215.4 + 226.5 + 206.6) / 3 = 216 mg ± 9.9 AA/100 g sample
The amount of ascorbic acid found in baobab fruit pulp was 216mg /100 g. This corresponds to a range of between 300 mg/100 g (Nour et al., 1980) and 150mg/ 100g (Manfredini et al., 2002). Therefore, according to Directive 2008/100/EC, it is possible to claim that the fruit pulp has “high” levels of ascorbic acid.
3.1.8 Acidity
The aim of the titration was to discover how much 0.1 M NaOH is required to record a pH
of 8.1 in the sampl e. This was achi eved by recordi ng the amount of NaOH requ i red to record a pH of j ust bel ow 8.1 and agai n for a pH of j ust above 8.1. The actual amount of NaOH required to record a pH of 8.1 was then be calculated as below.
49
Table 3.8: Titration results.

In order to calculate the actual volume of NaOH required to reach a pH of 8.1, at figures obtai ned from the ti trati on were used to create a l i ne usi ng pH on the y-axis and volume on the x-axis. The sl ope and i ntercept of the l i ne were then used to cal cul ate the final volume of titration requi red for pH 8.1.
Table 3.9: Titratabl e aci dity results: fruit pulp.

An average of the results was compi l ed: (5.41 + 5.42 + 5.28) / 3 = 5.37 % ± 0.08 aci di ty as ci tri c aci d (g/100g)
T i tratabl e acidity (TA) measures the total aci d content in a food or beverage system and i s determi ned by ti trati on of the aci ds in the food system wi th a standard base (Friedrich, 2001). Titratable acidity should not be confused with pH, as the latter is formally dependent upon the activity of hydrogen ions when measuring acidity or alkalinity. The acidity recorded, 5.37 %, may be a contri buti ng factor to the cl ai ms that the taste of the fruit pul p i s tart and aci di c.
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3.1.9 Anti-browni ng capacity
The purpose of thi s test was to determine if baobab fruit pul p coul d di spl ay functi onal

properti es in the food i ndustry, in terms of the inhibition of enzymi c browning of sl i ced appl es duri ng storage. The hi gh level s of ascorbi c acid observed in the sampl e may hel p to reduce the ri sk of enzymati c browning. The enzymi c browning of food i s undesi rabl e as i t results in the food bei ng rejected on by consumers on the basi s that the appearance i s unappeal i ng. Colour was anal ysed usi ng the H unter ® L a b Colour Scal e. In thi s scal e, the col our space i s represented in a cube form (figure 3.1). From the di agram, it can be seen that the L axis runs from top to bottom. The maximum L value i s 100 and corresponds to a perfect reflecting diffuser. The minimum L values is 0, corresponding to black. The a and b values have no numeri callimits. Positive aisred, whi ch negati ve a i s green. Positive b i s yel l ow, which negative bisblue.

Figure 3.1: H unter L a b col our scal e.
The results of the anti-browning capacity over 3 hours were i nterpreted and compi l ed in Figure 3.2 in order to provi de a vi sual representati on of the l evel s of browning of al l the sampl es i.e. the two control s (water di p and phosphate buffer solution di p), both baobab sampl es (i n water and phosphate buffer solution), the ascorbi c aci d sampl es (i n water and phosphate buffer solution) and the sampl es di pped in sodium metabi sul phite.
51

Delta E
16.00
14.00
12.00
10.00
8.00
4.00
0.00
6.00
2.00
0          1          2          3
Colour Change
Time Ihours)
Control-Water Control PBS Baobab 1% Water Baobab 1% PBS SMB
AA Water
AA Buffer
Figure 3.2: Overal l col our change over three hours i ll ustrati ng the affect of each solution on the browning of appl e sl i ces.
It can be cl earl y seen from the graph that the appl e sl i ces di pped in sodium metabi sul phi te di spl ayed the l east amount of browning. Sodium metabi pul phi te i s commerci al l y used in order to hel p prevent enzymi c browning, and thus extend the shel f-l i fe of a product. H owever, in recent ti mes, sodium metabi sul phi te has come u nder scruti ny because of the l i nk between possible si de effects in hu mans and the preservati ve.
The second most successful solution in thi s test that i nhi bited browning over 3 hours, in compari son to the control s, was the ascorbi c aci d in phosphate buffer solution. The purpose of the phosphate buffer solution was to hel p mai ntai n a constant pH in the solution, so as not to degrade the ascorbi c aci d, as ascorbi c aci d i s pH sensitive. W hen the resul t of ascorbi c aci d in buffer i s compared to the equi valent amou nt of ascorbi c aci d in water, i t can be seen that the buffer has an effect on the acti vi ty of ascorbi c aci d. This i s most l i kel y as a resul t of the fact that buffer creates a more desi rabl e envi ronment for the ascorbi c aci d, in compari son to water whi ch i s l i kel y to have caused the ascorbi c aci d to degrade sooner. T here i s l i ttl e di fference between the two control s, both in buffer and in water. The fact that appl e sl i ces di pped in buffer al one browned at the same rate as the appl e sl i ces di pped in only water, proves that the buffer itself has no effect on the apple browning. Therefore, it
52
can be stated that the buffer only protects the ascorbic acid and prevents its degradation, and al one has no effect on the browning.
As previ ousl y menti oned, there i s l i ttl e di fference between the ti mes taken for browning to occur in both control s. The baobab in phosphate buffer results in the same l evel of browning as the control s for hours 1 and 2. H owever, at hour 3, the baobab in phosphate buffer i ncreases browning more than the control. A si mi l ar reacti on can be seen with the ascorbi c aci d in water. At ti me 0 and after 1 hour, the browning i s si mi l ar to the control s. H owever, at hour 2 and 3 the appl es in thi s solution brown more rapi dl y than the control s.
The baobab in water solution caused the most browning out of al l the sampl es. This coul d be attri buted to the addi ti onal col our present in the solution as a resul t of the col our of the fruit pul p i tsel f. The presence of substrates in the baobab pul p extract (i.e. phenol i c compounds), that may favour the activity of polyphenol oxidase (PPO) enzymes in the appl e sl i ces, coul d al so expl ai n the i ncreased browning rate of sampl es treated as such.
It was concl uded that the baobab contai ned poor anti -browni ng properti es and shoul d not be appl i ed to the food i ndustry as a method for preservati on of foods in terms of anti – browning capacity.
3.1.10 Crude Extractions
In order to carry out anal ysi s of carotenoi ds, total phenol i c, f l avonoi d and tannin content and anti oxi dant capaci ty of the fruit, i t was necessary to carry out crude extraction of the fruit pul p, in order to extract both the hydrophi l i c and l i pophi l i c components of the fruit pul p.
Water was used to extract the water sol ubl e/hydrophi l i c components of the fruit pul p. This was carri ed out at two temperatures, room temperature and 90 °C, to determi ne if a di fference in temperature woul d enabl e a hi gher rel ease rate of the compounds in water. The resulti ng pellet formed after the hydrophi l i c extraction was subsequently extracted in order to obtai n the water insoluble components. A l ess polar sol vent than water was used to extract the components of the remai ni ng pellet after water extraction. A mixture of acetone and methanol was used to extract these fat sol ubl e/l i pophi l i c components.
53
D uri ng the extraction process, i t was important to avoi d exposure of the fruit pul p/extracts to l i ght, in order to prevent l i ght degradati on of the extracted components. This was achieved by wrappi ng foi l around the plastic tubes in whi ch the extracts would be stored.
During initial testing, it was found that the crude extracts that had been subjected to high temperatures (90 °C) had l ittle antioxidant activity and a low phenol ic, flavonoid and tannin content. This could be because ascorbic acid and other non-reducing substances such as phenolic compounds are heat sensitive. As mentioned previously, the purpose of using a heati ng step duri ng the extraction was to optimise the rel ease rate of the soluble substances in the fruit pul p. As a result of the fact that the pul p is so fine, it was found that there was no need to di srupt the cel l wal l materi al by boi l i ng water. It was therefore deci ded that onl y the room temperature extracts woul d be used for the remai nder of the testi ng.
3.1.11 Test for presenœ of carotenoids
C arotenoi ds are natural pigments whi ch are synthesi sed by plants and are responsi bl e for the bri ght col ors of vari ous fruits and vegetabl es, and have been i nvesti gated for thei r antioxidant properties (Paiva & Russell, 1999). A positive test for carotenoids would have resul ted in an orange col oured layer bei ng formed in the separati ng funnel . As there was no orange layer present, it can be stated that either the test for carotenoids was not successful, or there are no carotenoids present in the sampl e. If the former expl anati on i s to be fol l owed, retesti ng shoul d be carri ed out in order to rul e out the l i kel i hood of techni cal errors. If the latter expl anati on i s to be expl ored, namely the i dea that no carotenoids are present, thi s may be expl ai ned by the fact that the fruit pul p of the baobab i s encapsul ated in a hard shel l whi ch prevents the pul p from bei ng exposed to sunlight. This detai l may i ndi cate that the plant does not product carotenoids, as carotenoids are produced as plants respond to sunlight.
3.1.12 Total Phenolic Content
The phenol i c content was determi nd by the Fol i n C i ocal teu method and expressed as gal l i c aci d equi val ents mg per 100 mL (GA E mg/100mL ). The resul ts are shown in table 3.10.
54
Table 3.10: Total phenol i c content.
2072

The hydrophi l i c extract (H E) contai ns a hi gher phenol i c content in compari son to the l i pophi l i c extract (LE). This i s because phenol i c compounds are more soluble in water. The l i pophi l i c extract (L E) contai ns non polar compounds i ncl udi ng fatty aci ds. L ami en-M eda et al. (2008) found the total phenol i c content of baobab extracts to be 40.72 mg/g pul p, al most double the content achi eved in thi s experi ment. This di screpancy may be as a result of the fact that thi s group used di fferent sol vents and extraction processes. It can be seen from the results that the H E contri butes to al most six ti mes the content of phenol i cs, in compari son to the contribution of L E. This i s i ll ustrated by the col umn chart bel ow (Figure 3.3).
Figure 3.3: Contribution of LE and H E to the total phenol i c content.
3.1.13 Total Flavonoid Content Table 3.11: Total Flavonoid Content

mg GAE/g pulp
25
20
15
10
0
5
Hydrophilic Extract
Lipophilic Extract
55
It can be seen from the results that the hydrophi l ic extract (H E) contai ns a higher flavonoi d content, in compari son to the l i pophi l i c extract (LE). It can al so be noti ced that a higher phenol i c content was observed in compari son to the fl avonoi d content. This coul d be explained by the presence of other non-reducing substances, such as ascorbic acid. A ccordi ngl y, thi s effect was not noti ced for the L I, where ascorbi c aci d and possi bl y other i nterferences such as peptides are not present. The hi gh content of phenol i cs detected coul d be al so due to the presence of phenol i c aci ds, not detected wi th the fl avonoi d assay. The H E contri butes to more than four ti mes the cotent of fl avonoi ds, in compari son to the LE. This is i l l ustrated by the col umn chart bel ow (Figure 3.4).

mg CE/g pulp

15 10 5 0

Hydrophilic Extract Lipophilic Extract

Figure 3.4: Contribution of LE and H E to the total fl avonoid content. 3.1.14 Total Tannins/Procyanidins
Table 3.12: Total Tannin Content

From the results, it can be seen that the H E has a higher content of tannis, in compari son to the LE. As a resul t of the fact that procyani di ns are compounds that yi el d anthocyani di n pigments upon oxi dati ve cl eavage in hot al cohol s (i.e. via aci d butanol chemi stry), the presence of these compounds may be assessed by readi ng the absorbance. A maximum
56
absorbance at 549 nm confi rms the presence of the cyani di n agl ycone. It i s evi dent from the graph (Figure 3.5) bel ow that the readi ng obtai ned from the spectrometer peaks at 549 nm, thi s confi rmi ng the presence of tannins.
Figure 3.5: A bsorbance readi ng from 400-610 nm
The H E contri butes to more than twi ce the amount of tannins, in compari son to the LE. This i s i l l ustrated by the col umn chart bel ow (Figure 3.5).
Figure 3.6: Contribution of LE and H E to the total tannin content.
3.1.15 DPPH
The absorbance readi ngs of the D PPH assay, for both the hydrophi l i c extract (H E) and
l i pophi l i c extract (LE) were compi l ed in Excel . The absorbance readi ngs of the standards,
both in water and acetonejmethanol, were al so compi l ed. A cal i brati on curve was

Absorba nce (AU)

1 0.8 0.6 0.4 0.2 0

Series1

400 430 460 490 520 550 580 610
Wavelength (nm)

mg CyE/g pu I p
4
0
5
3
2
1
Hydrophilic Extract Lipophilic Extract
57
formul ated usi ng the standard absorba nce readi ngs for each sol vent (A ppendi x v). T hese cal i brati on curves were subsequently used to determi ne the mg AA E present in each gram of baobab fruit pulp.
DPPH (AEAC mg
Table3.13: DPPH results. 02

Figure 3.7: Contribution of LE and H E to the total D PPH content.
From the results, it was observed that the antioxidant capacity of the hydrophilic extract was hi gher compared to the l i pophi l i c extract. LE contai ned l ess than hal f the anti oxi dant capacity of the H E. The antioxidant capacity of the LE was probably due to the presence of fl avonoi ds al one, whi le ascorbic acid may contribute to the hi gher anti oxi dant capacity of the H E, in addition to fl avonoi ds. T hese fi ndi ngs suggested that despi te a lower amount of phenol i c antioxidants in the baobab pul p were of medium-l ow pol arity, they showed strong anti oxi dant capaci ti es, i ndi cati ng the i nterest in usi ng sol vents of lower pol ari ty than water for characteri si ng the phenolic profile of the fruit pul p.
3.1.16 FRAP
The absorbance readi ngs of the FRA P assay, for the sampl es and standards, were compi l ed
as di scussed in section 3.1.15. The cal i brati on curves used for FRA P can be seen in

mg AAE/g pu lp
12
10
8
4
0
6
2
Lipophilic extract Hydrophilic extract
58
A ppendi x iv. From the results in Table 3.14, it can be seen that H E has a more powerful antiFoxi dant capaci ty than LE.
Table 3.14: FRAP results.
1059 ± 172

Figure 3.8: Contribution of LE and H E to the total FRAP content.
The measurement of the antioxidant capacity of foods is a matter of growing interest because it may provi de information such as the anti oxi dant acti vi ty that the food may confer i nsi de the organi sm when i ngested (Z ul ueta et al., 2009). The anti oxi dant capacity of foods depends on many factors, i ncl udi ng the col l oi dal properti es of the su bstrates, the conditions and stages of oxi dati on, and the localisation of anti oxi dants in di fferent phases (Frankel & Meyer, 2000). M oreover, the measured antioxidant capacity of a sample depends on whi ch technol ogy and whi ch free radical generator or oxi dant i s used in the measurement (Zul ueta et al., 2009). Therefore, it i s important to carry out at l east two anti oxi dant assays, for exampl e D PPH and FRAP in order to compare and contrast results.
3.1.17 Antimicrobial assay
The results of the anti mi crobi al assay were compi l ed i nto graphs to provi de an i l l ustrated description of the antimicrobial activity of baobab fruit pulp. The concentration of baobab

mg CyE/g pu I p
40
50
30
20
10
0
Hydrophilic Extract Lipophilic Extract
59
(mg/mL) used in the assay was cal cul ated usi ng the initial dilutions of 1g/40mL for the hydrophi l i c extract, and 1g/50mL for the l i pophi l i c extract as starti ng points (A ppendi x i i i ). The anti mi crobi al effect agai nst Staphylococcus aureus of both the hydrophi l i c and l i pophi l i c baobab extracts can be seen in Figures 3.9 and 3.10 respectively.
Figure 3.9: A nti mi crobi al capaci ty of hydrophi l i c extract on Staphylococcus aureus.
Figure 3.10: A nti microbial capacity of l i pophi l i c extract on Staphylococcus aureus.
From the graphs above i t i s evi dent that the hydrophi l i c extract (H E) i nhi bi ted the growth of Staph. aureus more than the l i pophi l i c extract (L E). The concentration of H E that had the most apparent inhibition was at a concentration of 12.5 mg/mL. Concentrations of 6.25, 3.125 and 1.5mg/mL showed a l evel of inhibition. The concentration of 0.8 mg/mL had no
Hydrophilic Staphylococcus aureus

OD 600 nm	2 1.5 1 0.5 0			Staph Aureus 12.5mg/m L 6.25mg/m L 3.125mg/mL 1.5mg/mL 0.8mg/mL
		0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26

Time (hours)
Lipophilic Staphylococcus aureus

2
1.5
1
0.5
0
OD 600 nm
Staph Aureus 10mg/m L 5mg/mL 2.5mg/mL 1.25mg/m L 0.625mg/mL
0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26
2.5
Ti me (hours)
60
effect on the growth of Staph. aureus. In terms of the l i pophi l i c extracts, concentrations of 10 and 5 mg/mL exhi bi ted some inhibition of Staph. aureus, however the remai ni ng concentrations had no visible effect on the growth of the bacteri a. The H E most l i kel y caused more inhibition of Staph. aureus because of the hi gher content of phenol i c compounds contai ned in the H E in compari son to the L E, as di scussed in section 3.1.12.
The anti microbial effect agai nst Escherichia coli of both the hydrophi l i c and l i pophi l i c baobab extracts can be seen in Figures 3.11 and 3.12 respectively.
Figure 3.11: A nti microbial capacity of hydrophi l i c extract on Escherichia coli.
Figure 3.12: A nti microbial capacity of l i pophi l i c extract on Escherichia coli.
Hydrophilic Escherichia coli

	2 1.5 1 0.5 0
OD 600 nm						E. coli 12.5mg/m L 6.25mg/m L 3.125mg/mL 1.5mg/mL 0.8mg/mL

0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26
Ti me (hours)
Lipophilic Escherichia coli
Time (hours)

1.5
1
0.5
0
OD 600 nm
E. coli 10mg/mL 5mg/m L 2.5mg/mL 1.25mg/mL 0.625mg/m L
0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26
2
61
From these graphs, it i s evident that the hydrophi l i c extract (H E) i nhi bited the growth of E. coli more than the l i pophi l i c extract (L E). The concentration of H E that had the most apparent inhibition was at a concentration of 12.5 mg/mL. Concentrations of 6.25, 3.125 and 1.5mg/mL showed a l evel of inhibition. The concentration of 0.8 mg/mL had l ittl e effect on the growth of Staph. aureus. In terms of the l i pophi l i c extracts, concentrations of 10, 5, 2.5 and 1.25 mg/mL exhi bi ted varyi ng l evel s of inhibition of E. coli, however the concentration of 0.652 mg/mL had no visible effect on the growth of the bacteri a. It must be noted that the LE had greater i nhi bi tory powers agai nst E. coli than agai nst Staph. aureus. In fact, both the H E and LE di spl ayed stronger inhibition agai nst E. coli than Staph. aureus. As di scussed earl i er, the H E most l i kel y caused greater inhibition in compari son to the LE because of the hi gher content of phenol i c compounds contai ned in the H E.
The anti mi crobi al effect of the H E coul d be as a result of peptides and phenol i c compounds contai ned in the water extract. In order to determi ne the extent to whi ch each of these compounds are responsi bl e for the anti mi crobi al action, fracti onati on woul d need to be carri ed out, and the assays repeated wi th the fracti oned extracts. A l though studi es have previ ousl y concentrated on the anti mi crobi al acti vi ty extracts of the stem, l eaf and root of the baobab tree (M asol a et al., 2009; A nani l et al., 2000), i t i s cl ear from the results of thi s project that the fruit pul p does have some l evel of anti mi crobi al acti vi ty. The anti mi crobi al acti vity of the fruit pul p coul d be useful at i nhi bi ti ng the spoi l age of food products contai ni ng the fruit pul p.
3.1.18 Prebiotic assay
The resul ts of the prebi oti c assay were compi l ed i nto graphs to provi de an i l l ustrated description of the outcome of the assay. The concentration of baobab (mg/mL) used in the assay was cal cul ated as outl i ned in section 3.1.17. The prebi oti c effect exhi bi ted on Lactobacill us brevis of both the hydrophi l i c and l i pophi l i c baobab extracts can be seen in Figures 3.13 and 3.14 respectively.
62
Figure 3.13: Prebioti c acti vity of l i pophi l i c extract on Lactobacillus brevis.
Figure 3.14: Prebioti c activity of l i pophi l i c extract on Lactobacillus brevis.
From the graphs, i t i s evi dent that both the H E and LE extracts di d not support or i ncrease the growth of Lactobacillus brevis. Figure 3.13 displays high levels of growth between hours 1-11, correspondi ng to the H E at a concentration of 12.5 mg/mL . H owever, thi s concentration subsequently goes on to i nhi bit the growth of Lactobacillus brevis from

2
1.5
1
0.5
0
OD 600 nm
L. brev 12.5mg/mL 6.25mg/mL 3.125mg/m L 1.5mg/m L 0.8mg/m L
Hydrophilic Lactobacillus brevis
2.5
0 1 2 3 4 5 6 7 8 9 1011 1213 1415 16 17 18 19 20 2122 23 24 25 26
Time (hours)
Lipophilic Lactobacillus brevis
Time (hours)

2
1.5
1
OD 600 nm
0.5
0
2.5
0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26
L. brevus 10mg/mL 5mg/m L 2.5mg/m L 1.25mg/mL 0.625mg/m L
63
hours 15-24. In al l other H E extract concentrations, there extracts had no effect on the growth of Lactobacillus brevis. Figure 3.14 shows all LE concentrations inhibited the growth of Lactobacillus brevis. This was most l i kel y due to some resi dual acetone or methanol whi ch may not have been evaporated compl etel y in the LE preparati on step, as di scussed in section 2.1.17.
The prebi oti c effect exhi bited on Lactobacillus plantarum of both the hydrophi l i c and l i pophi l i c baobab extracts can be seen in Figures 3.15 and 3.16 respecti vel y.
Figure 3.15: Prebioti c acti vity of l i pophi l i c extract on Lactobacillus plantarum.
Figure 3.16: Prebi oti c acti vity of l i pophi l i c extract on Lactobacillus plantarum.
Hydrophilic Lactobacillus plantarum
Ti me (hours)

2
1.5
1
0.5
0
OD 600 nm
L. Plant 12.5mg/m L 6.25mg/m L 3.125mg/m L 1.5mg/m L 0.8mg/m L
0 1 2 3 4 5 6 7 8 9 10111213141516171819 20 2122 23 24 25 26
2.5
Lipophilic Lactobacillus plantarum
Ti me (hours)

2.5
2
1.5
1
0.5
0
OD 600 nm
L. plantarium 10mg/m L 5mg/mL 2.5mg/mL 1.25mg/mL 0.625mg/mL
0          5          10        15        20        25        30
64
Si mi l ar resul ts were achi eved with Lactobacillus plantarum, as with Lactobacillus brevis. It was noti ced that the H E at 12.5 mg/mL al so i nhi bi ted the growth of Lactobacill us plantarum, in a si mi lar manner to Lactobacillus brevis.
It i s cl ear that the crude extracts di d not confer prebi oti c acti vi ty. This may be due to a number of reasons, i ncl udi ng the fact that, duri ng the extraction process, fibre was not extracted. Fermentable fibres, such as ol i gosacchari des, have been attri buted to prebi oti c acti vi ty (G i bson, 2004; Rao, 2001; N apol i tano, 2009). The extraction of fibre requi res a hydrol ysi s step. The use of hydrol ysi s coul d be used in the future to i nvesti gate the prebiotic capacity of the pulp. Alternatively, the whole pulp could be fermented with probi oti c bacteri a in anaerobi c conditions, in order to refl ect the conditions in the intestine. The use of these approaches may result in prebi oti c activity.
65
Section 2: Product Development
3.2.1 Recipe development
The ingredients used for the formulation of the cereal bar were chosen for their health
benefits and ability to add to a “clean label” approach to the product, with no artificial or
unnecessary additives. As the product was ai mi ng to fal l i nto the category of cereal bar, there was a necessity for at least one cereal to be present in the recipe. Oats have a number of reported beneficial effects, both functionally wi thi n the food matrix and as a biologically functional in the body. In terms of functionality as an ingredient in the food matrix, oats provi de bul k and al l ow for absorbance of l i qui ds, thus faci l i tati ng bi ndi ng. Oats have al so been associated with a number of health benefits, including cholesterol lowering properties (Braaten et al., 1994), improving transit in the gut and prebiotic properties, the latter two improving gut condition and health. In addition to these benefits, oats have been proven to contri bute to sl ower glucose absorption (Jenki ns et al., 2004), resul ti ng in oats bei ng classified as a low GI food. These health benefits have been reported to be as a result of the soluble fibres present in oats, most notably β-glucan. It has been reported that soluble fibre­based prebiotics are becoming increasingly important as health-promoting functional foods (M acfarl ane et al., 2006). Taking these points i nto consi derati on, it was deci ded that oats would be used in order to fulfil this requirement of a cereal component in the recipe. Dates were chosen to be one of the main ingredients as a result of their sticky texture when bl ended. In addition to provi di ng sweetness to the bar and el i mi nati ng the need for added sugars/syrups, dates provi de a source of fibre, energy and vitamin B12, a vitamin which vegetari ans often l ack in thei r di ets. Dates are rel ati vel y expensi ve, so in order to reduce the amou nt of dates requ i red, raisins were i ncl uded in the reci pe. Raisins were fou nd to be an economi cal alternative to usi ng only dates in the reci pe, and have si mi l ar bi ndi ng characteri sti cs when bl ended. The raisins al so add sweetness due to the hi gh content of naturally occurring sugars present in the fruit. It was decided that mixed nuts (including hazel , pecan, brazi l and wal nuts) woul d provi de essenti al fatty aci ds and fl avou r, as wel l as addi ng a crunchy texture to the bar. It was decided that appl e j uice would be added in order to provide a liquid portion to bind the mixture together, as well as adding natural sweetness. In addition to these ingredients, the baobab fruit pup was added in order to boost the
66
vitamin C and mineral content of the mixture. Various natural flavourings were considered that would complement the flavour of the baobab fruit, and are discussed in section 3.2.2.
Two methods of processing were considered for the cereal bar. The first was a method of cold pressing the bar, which i nvolved applyi ng pressure to the bar at room temperature in order to shape and com press the mixture. The second method applied to the bar was baking. This method involved placing the mou lded mixture into an oven for 15 minutes at 140 °C. Both methods were used at the reci pe devel opment stage, and a focus group was uti li sed in order to determi ne which method was more appeali ng to prospective consumers. An image of both the baked and cold pressed versions that were presented to the focus group can be seen in Figure 3.17. The focus group concluded that the texture of the cold pressed cereal bar was not appealing, and was likened to “raw dough”. This method of processi ng was subsequently rejected and the baking method, which received positive feedback, was uti lised for the remainder of the product development process.

Figure 3.17: Example of the effect of each of the processing techniques; the baked sample on the left and the cold pressed version on the right.
It was deci ded that each bar woul d be desi gned to wei ght 42 g. This figure was chosen from viewi ng the cereal bars currently on the market, and their composition. Table 3.15 provi des detai l s on cereal bars currentl y on the market, and thei r approxi mate wei ghts. An average of these bars was cal cul ated, and thi s figure was used for the remai nder of the product devel opment process.
67
Table 3.15: Cereal bar weights. Weight of cereal bars currently on the market, and an average of the wei ghts.

3.2.2 Addition of flavours
A focus group was once agai n util ised in order to provide feedback on the various flavours
as detailed in section 2.2.2. The majority of the flavours received positive feedback. H owever, the vani l la fl avouri ng was rejected by the group. As a result, the va ni l l a fl avouri ng was omi tted for the future devel opment, whi l e each other fl avouri ng was carri ed through to the sensory anal ysi s stage.

Figure 3.18: Flavoured cereal bars. Cereal bars containing several flavours including coconut, ci nnamon, cocoa and vani l la.
68
3.2.3 Prototype development for sensory analysis
The inclusion of baobab fruit pul p in the cereal bar reci pe for the purpose of sensory analysis was not permitted on ethical grounds. To overcome this obstacle, an alternative cereal bar was developed specifically for the purpose of sensory evaluation. This recipe included all other ingredients present in the original formulation, with the exclusion of baobab fruit pul p. Enough sampl e for 25 j udges was prepared.
3.2.4 Sensory Analysis 3.2.4.1 Paired preference test
The results of the paired preference test (Appendix vii) were statically analysed as illustrated in Table 3.16. The results show that the panel preferred the appearance of the samples that di d not contai n oats on top. As the number of agreei ng j udgements was greater than 32 which corresponded to a p value of 0.001, the results were said to be very highly significant. As a consequence of these results, the final product did not contain oats on top.
Table 3.16: Two-tai l ed stati stical anal ysi s of Paired Preference Test

8
A two-tailed datawas used becausethe resultswerebased on
preference.Theref ore, the 30results are very highly significant. These results indicate that the panel preferred the appearance of the cereal bar without oats on top.
3.2.4.2
Ranking
Testfor
Preference-Appearance
Ranking tests are often used to screen one or two
“bestsamples”
from a group ofsamples, rather than to thoroughly test all samples. However, no indication of themagnitude of di ff erence
between samples
i s obtai ned
becausesamplesare
onl yeval uated
in
rel ati onshi p
to each other. Theresults of thistest (Appendix
vii)
showed that thepanel preferred the
appearanceof the
sampl econtai ni ng
coconut.
I t
was noted by several
panel li sts however
that there was “not much difference” visually between most of the samples. The sample
69

wi th the l east preferred appearance was the sampl e contai ni ng cocoa. A nu mber of panel li st
commented that the appearance of this sample, stating that the colour was “too dark” and looked “burned compared to the other bars”.
3.2.4.3 Ranking Test for Preference-Aroma
The results of this test (Appendix vii) showed that the panel preferred the aroma of the
sample containing cinnamon. A panel l i st commented that the aroma of the sample
containing cocoa was “overpowering”. The least preferred sample in the aroma test was the
plain sample with no added flavouring.
3.2.4.4 Ranking Test for Preference-Flavour
The results of this test (Appendix vii) showed that the panel preferred the flavour of the sample contai ni ng cinnamon. Panel l i sts commented that the sample containing cinnamon
was “the best overall” and had a “great flavour”. Once again, the sample containing cocoa
was least preferred.
Taking all these results into account, it was decided that the added flavouring would be ci nnamon.
3.2.5. Final batch preparation
The final batch preparation was formulated to reflect the sensory data obtained, as di scussed in 3.2.4. The final batch di d not contai n oat flakes on top of the bar. The flavouring added to the final batch was ci nnamon. Ci nnamon has been l i nked to i mprovi ng bl ood glucose l evel s in patients with di abetes. In a 2003 study, it was found that as l ittl e as 1 g of cinnamon per day reduced serum glucose, triglyceride, LDL cholesterol, and total chol esterol in people with type-2 di abetes. The study suggested that the inclusion of cinnamon in the diet will reduce risk factors associated with diabetes and cardiovascular diseases (Khan et al., 2003). This information, coupled with the fact that there is no added sugar/gl ucose syrups in the cereal bar, may make thi s product an option for i ndividual s who suffer from diabetes, in addition to individuals who simply wish to control their blood glucose l evel s. As the control di d not contai n baobab, l ess appl e j u i ce was added to the
70
control in compari son to the other batches contai ni ng baobab. This alteration was carri ed out in order to mai ntai n a si mi l ar consi stency in al l the cereal bars. It woul d not have been appropri ate to add the same amount of apple j ui ce to the control as was added to the baobab product, as this would have resulted in a product with a an entirely different texture to the baobab product. It was noted however that the control had l ess appl e j ui ce i.e. moi sture, and this may affect the results when both products were tested further in the project. The bars were vacuum packed in order to mai ntai n freshness. The bars were stored in the dark, in a cool dry place in order to prevent deteri orati on of the product over ti me, and al so to refl ect the storage parameters of the product if i t was to be stocked in a shop.
3.2.6 M oisture content
The importance of moi stu re content in foods has prev iously been discussed in section 3.1.1. The percentage moisture content of baobab fruit pulp was calculated using the following equation: M oi sture content (%) = (W1 – W2) / W1 x 100
Where: W1 is the weight of the initial sample, W2 is the weight of the sample dried at 105
°C.
Table 3.17: Moisture content: cereal bar.

Average moisture content of baobab cereal bar: (20.03 + 17.45 + 19.62) / 3 = 19.03 % ± 1.39
The average moi sture content of the cereal bars contai ni ng baobab was fou nd to be 19.03 % ± 1.39. D i screpanci es between the percentage moisture of baobab batches may be as a result of the portion of each batch sampled, i.e. a portion with more nuts may contribute less to moisture than portion containing a larger amount of oats with absorbed apple juice. This may be an explanation as to why the standard deviation of the results is relatively
71
hi gh. The moi sture content of the bar wi th no added baobab (control) was 18.72 %, whi ch
i s l ower than that of the bars wi th baobab. This i s most l i kel y as a resul t of the fact that the formulation of the control di d not contai n as much appl e jui ce as the other bars.
3.2.7 Ash Content
The methodol ogy and importance of ashi ng in foods has previ ously been di scussed in
section 3.1.2. The ash content was expressed as: Ash % = (W 1 / W 2) x 100 Where W1 = weight of ash and W2 = initial weight of dried sample at 105 °C Table 3.18: Ash content: cereal bar.

Average ash content of baobab cereal bar: (2.47 + 2.47 + 2.53)/ 3 = 2.49 % ± 0.03
The ash content of the cereal bars contai ni ng baobab was found to be 2.49 % ± 0.03. This figure contrasts greatl y to the figure obtai ned for the control bar whi ch was cal cul ated to be 2.01 %. The control contai ns al most 20 % l ess ash in compari son to the baobab cereal bar. This result i l l ustrates the fact that the addition of baobab fruit pul p i ncreases the mi neral content of a product. Therefore, it can be stated that baobab fruit pulp can be added to food products in order to forti fy a standard food product and thus i ncrease the nutriti onal content.
3.2.8 Water activity
Water acti vi ty (A w) i s consi dered as an index of the macroscopi c state of water in food. Its value affects the rate of vari ous processes in foods such as oxi dati on, browning, enzyme reacti ons and growth of mi croorgani sm (D erossi et al., 2006). In terms of mi croorgani sm growth, water acti vi ty i s a useful method of determi ni ng the l evel of water avai l abl e to mi croorgani sms i ncl udi ng bacteri a, moul ds and yeasts.
72
Table 3.19: Water acti vity l evel s of the cereal bars, on day 1 and day 8.

It can be seen from the results in Table 3.19 that the water activity for the control is lower than that of the baobab cereal bar. This difference coul d be expl ai ned by the fact that l ess appl e j ui ce was used in the formulation process for control, in compari son to the sampl es contai ni ng baobab. Table 3.15 i ll ustrates the A w l evel s requi red for the growth of a number of mi croorgani sms. By consulti ng the results of the A w test i n Tabl e 3.19, it can be seen that the maximum A w of al l sampl es was 0.79. W hen the water acti vi ty chart (table 3.20) i s
2          5
consi dered, i t can be seen that the maj ori ty of the mi croorgani sms are i nhi bi ted at thi s l evel
079      2          0726
of Aw. According to the water activity chart, the only growth that may occur in the bars is
0782    3          0733
± 0.031           Average 0.737 ± 0.014
that of yeasts, moulds and fungi. It can therefore be assu med that pathogeni c bacteri a wi ll
79        Cn
be i nhi bi ted at the l evel s of A w present in the cereal bars. H owever, for the purpose of mi crobi al safety, i t shoul d never be taken for granted that l ow A w l evel s rul es out the possi bi l i ty of the presence of pathogens. M i crobi al testi ng shoul d be carri ed out in order to confi rm the l ack of a presence of pathogens in the food.
73
Table 3.20: W ater acti vi ty values and associ ated mi crobi al spoi l age. (Z euthen & B ogh­Sorensen, 2000)

The purpose of testing the Aw levels of the bars at day 1 and again at day 7 was to 0.91-0.86
determine if time had an influence on the Aw of the samples. This is an important factor to of
s aureus           0.86
consider when determining the shel f-l i fe of the final product. The resul ts of the A w over ti me (Table 3.19) were used to determine if there was a significant difference in the Aw over 086-080
bacter  8075
seven days, i.e. if time i nfl uenced the A w of the sampl es. The analysi s of variance (A N OV A) was cal cul ated usi ng a software package (StatG raphi cs ®), and the results are i ll ustrated in Table 3.21.
74
Table 3.21: A nal ysi s of variance for A w.

The F-ratio, whi ch in thi s case equal s 2.38221, i s a ratio of the between-group esti mate to the withi n-group esti mate. Si nce the P-value of the F-test i s greater than or equal to 0.05, there i s not a stati sti cal l y si gni fi cant difference between the mean A w from one set of days to another at the 95.0% confidence l evel . T herefore, it can be stated that the nul l hypothesi s (H 0) can be accepted, i.e. ti me does not have an effect on the A w
3.2.9 M ineral content
The resul ts of the mi neral anal ysi s of the cereal bars can be seen in Table 3.22. W hen the
cereal bars contai ni ng baobab are compared with the control, it can be seen that in inclusion of baobab fruit pul p i ncreases the overal l mi neral content of the bar.
Table 3.22: Content (mg/100g sample) of calcium, sodium, magnesi um and potassium present in the cereal bars.

The col u mn chart (Figure 3.19) i l l ustrates the i ncreased l evel s of the four mi neral s that baobab confers on the bar. The most significant increase with the addition of baobab was seen in the potassium, where l evel s i ncreased by 34 %. A l though the calcium l evel s di d increase with the addition of baobab, it was thought that rise would have been more
75
si gni fi cant than was actual l y percei ved. An i ncrease of onl y 2 % of the overal l calcium content was achi eved, in compari son to the control.
Figure 3.19: M i neral content compari son. Compari son of calcium, sodium, potassium and magnesi um contents (mg/100g sampl e) in baobab cereal bar and control.
3.2.10 Shelf-life evaluation: ascorbic acid (AA)

The purpose of tracki ng the AA content over 7 days was to determi ne if ti me/storage had an effect on the l evel s of AA in the cereal bar. As one of the ai ms of the product devel opment i s to be able to cl ai m that the bar i s hi gh in vi tami n C, it i s important to ensure that the AA content does not decl i ne over storage. Cal cul ati ons were carri ed out in the same manner as outlined in section 3.1.7. Results of AA content at day 1 can be viewed in Table 3.23. In these results it can be observed that the content of AA in the baobab bars i s over 9 ti mes the l evel contai ned in the control. It must be noted that some of the vi tami n C contai ned in the baobab bars may be attri buted to the sl ightly hi gher volume of apple jui ce contai ned in the baobab bars. H owever, the majority of the i ncrease of AA in the baobab bar can be attri buted to the addition of baobab, whi ch has a consi derabl e effect on the l evel s of A A in the bar in compari son to the control. The results of AA content at day 8 can be viewed in Table

3.24.

120
100
80
40
60
20
0
Comparison of mineral content (mg/100 g sample)
in bar with baobab to control
Ca       Na       K         Mg
With baobab Without baobab
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Table 3.23: Ascorbic acid content day 1.

Table 3.24: Ascorbic acid content day 8.

It i s noted that the l evel of ascorbi c aci d has not been consi derabl y effected over storage from day 1 (10.959 ± 0.269) to day 8 (10.84 ± 0.26), with a decrease of an average of just 1 %. D etai l s of the percentage decrease can be seen in table 3.25, and are i l l ustrated in a col umn chart bel ow (Figure 3.20)
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Table 3.25: Percentage decl i ne in l evel s of ascorbi c aci d over 7 days

Figure 3.20: Illustration of the decl i ne in l evel s of ascorbi c aci d from day 1 to day 8.
The effect of time on A A l evel s was stati sti cal l y anal ysed, to determi ne if there was a si gni fi cant di fference between the l evel s of A A contai ned in the cereal bar at day 1 in compari son to day 8. The resul ts of the stati sti cal anal ysi s can be vi ewed in Table 3.26. The figures were assessed in terms of anal ysi s of variance. The F -ratio, whi ch in thi s case equal s 0.294359, i s a ratio of the between-group esti mate to the wi thi n-group esti mate. Si nce the P-value of the F-test i s greater than or equal to 0.05, there i s not a stati sti cal l y si gni fi cant difference between the mean AA from one peri od of time to the other at the 95.0% confidence l evel . The nul l hypothesi s H 0 may therefore be accepted, as a resul t of
Decline in levels of ascorbic acid over 7 days

11.5 11 10.5 10 9.5 9 8.5 8 7.5
			mg AA/100 g sample Day 1 mg AA/100 g sample Day 8
	1          2          3

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the fact that there is no stati sti cal difference in AA levels of the cereal bars from day 1 to day 8.
Table 3.26: Analysis of Variance for AA.

Two main mechani sms that can cause AA l oss in food i ncl ude degradati on by oxi dati on during processing and storage, and release by diffusion during processing such as baking (Arroqui et al., 2002). It is important to monitor levels of ascorbic acid in a product over ti me, to ensure that the l evel s do not degrade at a rapi d rate. AA l oss over storage can be reduced by sel ecting the appropri ate packaging and storage parameters. As ascorbic acid is light and heat sensitive, packaging and storage conditions should aim to reduce the exposure of a product to these two factors. Light exposure can be limited by using a packaging that covers the enti re product and thus blocks out the l i ght. Examples i ncl ude cardboard, foi l and col oured plastics. Certain storage conditions can be recommended in
order to reduce exposure to heat. Storage guidelines such as “store in a cool, dry place” can
be recommended on packaging, to prevent the consumer from subjecting the product to excess heat sources.
In order to clai m that the cereal bar i s a “source of” or “high in” vitamin C, it must contain at least 15 % or 30 % of the RDA of vitami n C respectively (FSAI, 2010). This pertai ns to 9 mg and 18 mg per serving, respectively. The current content of AA in the cereal bar is 10.84mg ± 0.26 per 100g sample. This relates to 4.55 mg per 42 g cereal bar. Therefore, the content of baobab fruit pulp is the cereal bar should be doubled in order to claim a “source
of” vitamin C in the product. This would involve increasing the content of baobab in the formulation from 5 % to 10 %. In order to claim that the bar was “high in” vitamin C, the level would need to be rai sed to the formulation contai ni ng 20 % baobab.
79
3.2.11 M icrobial content
The results of day 1 plate counts are illustrated in Tables 3.27 (PCA) and 3.28 (PDA). The
results are represented using the fol lowing decodes: x = no growth and y = growth. In all cases where growth was present, the colonies were too numerous to count (TNTC)
Table 3.27: PCA plate count day 1.

Table 3.28: PDA plate count day 1.

From these results it can be seen that contamination occurred. This was most li kely because of poor technique and contamination of sampl e duri ng preparati on. The test was repeated, usi ng further dilutions i.e. up to 10-8.

80
The results of day 5 plate counts are illustrated in Tables 3.29 (PCA) and 3.30 (PDA). The results are represented usi ng the fol lowi ng decodes: x = no growth and y = growth. The number of colony forming units (CFU) is represented by either TNTC or a figure representi ng the number of colonies present.
Table 3.29: PCA plate cou nt day 5.

As the results for the bars with added baobab state that the dilutions at 10-2 resulted in a microbial count of between 30-300, this dilution was used to determine the microbial load of the bulk dilution, and subsequently 1 g of cereal bar.
Cal cul ati ons:
CFU‟s at 10 -2 = (50 + 34 + 55)/3 = 46 CFU‟s average
When the dilution factor is considered, this figure must be multiplied by 102 in order to represent the mi crobi al l oad of 1 g of sampl e in 20 mL peptone, i.e. 46 x 102 = 4,600 or 4.6 x 103. This must then be divided by 0.1 (the amount of sample preparation spread per plate) i.e. (4.6 x 103)/0.1 = 4.6 x 104. This figure must then be multiplied a further time by 20, in order to account for the dilution with 20 mL peptone during the sample preparation steps, i.e. 4.6 x 104 x 20 = 920, 000 or 9.2 x 105 CFU‟s per 1 g sample.

CFU‟s at 10 -2 = (30 + 41 + 25)/3 = 32 CFU‟s average

= (32 x 102)/0.1 x 20 = 640,000 or 6.4 x 105 CFU‟s per 1 g sampl e.

81
B3: CFU‟s at 10 -2 = (26 + 30 + 37)/3 = 31 CFU‟s average = (31 x 102)/0.1 x 20 = 620,000 or 6.2 x 105 CFU‟s per 1 g sample.
As the results for the control state that the dilutions at 10-1 gave a microbial count of between 30-300, thi s dilution was used to determi ne the mi crobi al l oad of the bul k dilution, and subsequently 1 g of control.

XB: CFU‟s at 10 -1 = (175 + 192 + 163)/3 = 176 CFU‟s average

= (176 x 101)/0.1 x 20 = 352 000 or 3.52 x 105 CFU‟s per 1 g sample
In the case of PCA at day 5 (Table 3.29), the microbial growth in the control was lower than the growth in the bars with the added baobab. This may indicate that the baobab had its own mi crobi al l oad. This may have been eradi cated through a pre-processi ng step such as exposure of the fruit pulp to UV light in order to eliminate the microbes present. It must be noted that the identity of the microbes present was not determi ned, therefore it cannot be stated if the bacteria present were pathogenic or non-pathogenic.
Table 3.30: PDA plate count day 5. Plate count of PDA usi ng eight dilutions, where x = no growth, y = growth. Number of col ony forming units (CFU) is represented either by TNTC or a figure representi ng the number of colonies present.

82
As the results for both the control and the bars with added baobab state that the dilutions at 10-1 resulted in a microbial count of between 30-300, this dilution was used to determine the microbial load of the bulk dilution, and subsequently 1 g of cereal bar.
Cal cul ati ons:
CFU‟s at 10 -1 = (29 + 32 + 43)/3 = 35 CFU‟s average = (35 x 101)/0.1 x 20 = 70,000 or 7 x 104 CFU‟s per 1 g sample.
No growth of between 30-300 colonies, therefore there is said to be no microbial count.
No growth of between 30-300 colonies, therefore there is said to be no microbial count.
XB: CFU‟s at 10 -1 = (194 + 201 + 157) / 3 = 184 CFU‟s average
= (184 x 101)/0.1 x 20 = 360,000 or 3.6 x 105 CFU‟s per 1 g sample
In the case of PDA at day 5 (Table 3.30), the microbial growth in the control was higher than the growth in the bars with the added baobab. This could i ndicate that the presence of baobab inhibited the growth of yeasts and moulds, which did not occur in the control. It could also be as a result that the control was contaminated with moulds/yeasts during processi ng whi le the baobab bar was not.
One of the methods that could be used to determi ne the types of microbes present would be to use selective agar. This would characterise the microbes present in the samples, and would provide more detai led information of microbes present. Common microbes found in food often include coliforms, Escherichia coli (E. coli), Salmonella, Yeasts and Moulds, Bacillus cereus, Clostridium perfringen and Staphylococcus aureus. Selective tests could be carri ed out for each of these microbes, in order to determi ne thei r presence/absence. A
guideline to the limit of CFU‟s of each of these microbes that can be present in fortified
bl ended foods is i l l ustrated in Table 3.31. U si ng the sel ecti ve agar method, i t would be possible to determi ne if there were any pathogeni c bacteri a present. If pathogeni c bacteri a




83
were in fact present, it woul d be important to i mpl ement a strategy to provi de detai l s of where, in the development process, the contamination occurred. Steps could then be taken to prevent thi s form of contamination from occurri ng.
Table 3.31: Maximum number of CFU‟s permitted in fortified blended foods. (United Nations World Food Programme, 2009)

3.2.12 Shelf-life evaluation: colour
The purpose of assessing the colour over time was to determine if time had a significant
effect on the col our/appearance of the cereal bar. Appearance is an important attri bute in
terms of sensory appeal. If the colour of the product changes significantly over storage, it 100/g
may result in consu mers rej ecti ng the product over ti me. Sensory appeal can be a deci di ng
factor in terms of acceptabi l ity. Even if a product i s safe to eat and contai ns the same
nutritional content as stated, if it does not appeal to the senses of consumers, it will be
u          0/g
rej ected. The i nterpretati on of the H unter colour chart has been di scussed previ ousl y in
section 3.1.9. A table of results of the col our evaluation can be viewed in Appendix viii. 10/g
These figures were compi l ed to create a col umn chart, in order to ill ustrate the change in col our over time, from day 1 to day 8 (Figure 3.21). The chart clearly shows a decrease in the L* value, no change in the a* value and an i ncrease in the b* value.
84

45
40
50
35
30
25
20
15
10
0
5
Change in L*a*b* values over 7 days
L*        a*        b*
Series1
Series2
Figure 3.21: L *a*b* values over seven days.

Figure 3.22: Col our Plot cereal bar.
The col our plot (Figure 3.22) generated by the col ori meter i ll ustrates the change in col our over ti me, with day 1 on the l eft and day 8 on the ri ght of the figure.
85The detai l s of the col our testi ng were anal ysed usi ng the anal ysi s of variance, in order to determine if time had a significant effect on the colour of the bars over ti me. The results can be seen in Table 3.32 bel ow.
Table 3.32: A nal ysi s of Variance for col our

The P-val ues test the stati sti cal si gni fi cance of each of the factors. Si nce the P-val ue i s l ess than 0.05, it can be deduced that time does not have a stati sti cal l y si gni fi cant effect on col our at the 95.0% confidence l evel . This resul t i ndi cates that the nul l hypothesi s (H0) i s accepted, and that there i s no difference in the col our of the sampl es over time. This resul t provi des the information requi red to state that the cereal s wi ll not be rejected in sensory tests, as there i s no si gni fi cant di fference between the bars over storage.
3.2.13 Packaging, marketing, nutritional information and costings of final product 3.2.13.1 Packaging
Packaging: The final product was vacuum packed. Vacuum packi ng i nvol ves removi ng air from the packaging that surrounds the food and preventi ng i ts return by an ai rti ght seal . This method of packaging al lows for optimum freshness, as the product cannot be oxidised by oxygen present in air. Oxidation could result in the ranci dity of fats present in the nuts of the cereal bar. Oxi dati on al so destroys ascorbi c aci d.
Removi ng air from product packaging changes the micro-envi ronment surroundi ng the food, i nhi biti ng the growth of the majority of spoi l age organi sms and pathogens that grow in the presence of oxygen. The absence of oxygen in thi s form of packaging coul d i nhi bit the growth of aerobi c bacteri a, such as Staphylococcus sp., Streptcoccus sp., Enterobacteriacae sp. and Pseudomonas aeruginosa. Removi ng oxygen surroundi ng the food does however create conditions suitable for the growth of anaerobic organisms. The
86
most significant of these, from a public health perspective, is Clostridium botulinum. Regular testing should be carried out in order to ensure the absence of Clostridium botulinum and the spores. Another advantage of vacuum packaging i ncl udes the fact that it can prevent moi st foods from dryi ng out, as wel l as mai ntai ni ng flavour for longer.
As well as being sensitive to oxygen, ascorbic acid is light sensitive. The theoretical packi ng of thi s product woul d consi st of a col oured plastic depi cti ng a design i ncl udi ng the logo, name of the product, i ngredi ents list and images that woul d block the light out, therefore preventing the light-induced degradation of the vitamin C. A rough draft of the packaging can be seen below in Figure 3.23.

Figure 3.23: Packaging, front cover.
The i magery used for the packaging as i l l ustrated above in Figure 3.23 was chosen because it contains a depiction of oats, which is one of the ingredients contained in the cereal bar. The inspiration for the colour scheme originated from colour of the fruit pulp, as it consists of a light orange colour. The colour scheme appeals to both males and females, and should not alienate either sex in terms of a target market. The significance of oats as an ingredient
is hi ghl i ghted in the name “OatiVita”. This name not only reflects the presence of oats in
the reci pe, but al so ai ms to conjure an image of a heal thy l i festyl e by the inclusion of
“vita”. As vita is the Italian word for life, it was decided that this name reflected well both
the ai m that the bar shoul d contri bute towards a heal thy di et and li festyl e, in addition to
87
incorporating an element of one of the colloquial names of the baobab tree, “Tree of Life”.
The back cover of the packaging, as i l l ustrated in Figure 3.24, contai ns details of the product, i ncl udi ng i ngredi ents, nutritional information, al l ergy advi ce, best before date, net
weight and manufacturer‟s details, in order to inform the consumer as to the contents of the
product. This section of the label also contains additional details that the health conscious
consumer may find of interest, including a “free from” section, outlining details such as no
added sugar, preservatives, col ours or flavouri ngs. If the content of baobab was i ncreased to
10 %, as di scussed in section 3.2.10, the label could include the term “source of vitamin C”.
Figure 3.24: Packaging, back cover. 3.2.13.2 Marketing
This product coul d be theoreti cal l y marketed to appeal to a wi de range of i ndi vi dual s. As well as the fact that it contai ns no additives, preservatives, col ouri ngs, added sugar or yeast, the product al so qualifies to cl ai m a range of benefi ts, i ncl udi ng:
Sui table for vegetari ans
Suitable for coeliacs,
Lactose free
Low GI
Source of Vitamin C (when i ncreased to 10 % content)

Oat, nut and fruit bar with added baobab fruit.
Ingredients: Dates, oats, mixed nuts (hazel, pecan, brazil and walnuts), raisins, apple juice, baobab fruit pulp, flavouring.
Suitable for: Vegetarians, coeliacs. Low GI. Source of fibre. Allergy advice: Contains nuts.
Best before:     Net weight: е42 g         Manufacturer’s details:
Nutritional Information per bar (42g): Energy 134 kcal, protein 2.4g, carbohydrate 24.7g, fat 4.6g, fibre 2.3 g, sodium 0.006g, calcium 46.5 mg.
Free from: Gluten, lactose, added sugar, yeast, artificial flavouringsi preservatives/colours.

88

High in fibre

3.2.13.3 Nutritional Information
The nutri ti onal content of the cereal bar was cal cul ated by gatheri ng the nutri ti onal content per 100 g of the component i ngredi ents. The amount of these i ngredi ents requi red to produce 100 g and 42 g respectively of the bar was determined. The figures were then compi l ed and tabul ated i nto Table 3.33. The cal cul ati ons were carri ed out in Excel, and can be vi ewed in A ppendi x ix.
Table 3.33: N utri ti onal information.

89
The nutritional content of the cereal bar would be appealing to individuals who wish to mai ntai n a l ow calorie diet, as the cereal bar contai ns onl y 6.7 % of the RDA of calories, based on a 2000 calorie/day diet. The bar contains almost 5 % of the RDA of protein. The bar contains the following percent of the RDA of the named minerals: 6 % calcium, 11 % magnesi u m and 1 % potassium. As a resu l t of the fact that the bar contai ns 5.5 g fibre per 100 g, it can be claimed that the bar is a “source of” fibre (FSAI, 2010). This cl ai m coul d be promoted to “high fibre” with an increase of just 0.5 g fibre per 100 g. As the bar
contai ns 0.006 g of sodium, equi val ent to 0.015 g salt, i t contri butes to j ust 0.4 % of the RDA of salt for adults.
3.2.13.4 Costing estimation
The cost per unit of the product was calculated using the price of the contributing i ngredi ents. The method used to esti mate the cost of the product can be seen in table 3.34.
Table 3.34: Cereal bar cost estimation.

Costing for product only:

Total cost per bar: €0.262 ≈ €0.26, total cost per kg: €6.2381 ≈ €6.24. This figure could be dramatically reduced if buying wholesale. Basic costing should also take into account the cost of packaging, labour, transport, storage and a number of additional factors.
90
91
C hapter 4 Conclusion
92
At the end of thi s study, the fol l owi ng conclusions can be made:
The successful characterisation of baobab fruit pulp has been achieved using a number of analytical techniques. This is proven by the fact that that the content of moi sture, ash, fat, protei n, pecti n, mi neral s, ascorbi c aci d and acidi ty were determi ned.
Baobab fruit pulp does not have an anti-browning effect on the enzymatic browning of sl iced apples over ti me.
Antioxidant assays, such as FRAP and DPPH, demonstrated that baobab fruit pulp contains high levels of antioxidants. In addition to this, the fruit pulp contains high l evel s of phenol i cs and fl avonoi ds.
Crude water and acetone/methanol extracts of baobab fruit have antimicrobial properties, but do not have prebiotic properties under aerobic conditions.
Sensory analysi s determi ned that the baobab cereal bar shoul d contai n ci nnamon as an added flavouring, and should not contain oat flakes on the top surface of the bar.
The ascorbic acid content, water activity and col our of the cereal bar did not change significantly (p≥0.05) over storage.
The addition of baobab fruit pulp increases the mineral content of a product considerably.

93
94
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Afolabi O. R.; Popoola T. O. S., (2004), The effects of baobab pulp powder on the micro flora involved in tempe fermentation. Eur Food Res Technol, 220, 187-190. Almroth S.; Latham M., (1995), Rational home management of diarrhoea. PubMed, 345, 709-711.
Al-Qarawi A. A.; Al-Damegh M. A.; El-Mougy S. A., (2003), Hepatoprotective Influence of Adansonia digitata Pulp Journal of Herbs, Spices & Medicinal Plants, 10, (3), 1-6.
Ananil K.; Hudson J. B.; Souzal C. d.; Akpaganal K.; Tower G. H. N.; Amason J. T.; Gbeassor M ., (2000) Investigation Of M edi ci nal Plants Of Togo For Antiviral And Antimicrobial Acti vities. Pharmaceutical Biology, 38, (1), 40-45.
Arnold T. H.; Wells M. J.; Wehmeyer A. S., (1985), Khoisan food plants: taxa with potential for future economic exploitation. Plants for arid lands, 69-86.
Arroqui, C., Rumsey, T. R., Lopez, A., & Virseda, P., (2002), Losses by diffusion of ascorbic acid during recycled water blanching of potato tissue. Journal of Food Engineering, 52(1), 25-30.
Association of Analytical Communities, Official Method 967.21 Ascorbic Acid in Vitamin Preparations and Juices, http://www.aoac.org/omarev1/967_21.pdf, accessed 11/04/2011
Atawodi S. E., (2005), Comparative in vitro trypanocidal activities of petroleum ether, chloroform, methanol and aqueous extracts of some Nigerian savannah plants. African Journal of Biotechnology, 4 (2), 177-182.
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101
Appendices
102
Appendix i
Stock solutions: (1000 ppm)
NaCl: To make 1000ppm (1g/ L) Na+ from NaCl Formula: Na+ + Cl- = x
N a+
-Y 22.990 + 35.453 = 2.5419 g/L
22.009
Dissolve 2.5419 g of NaCl in 1 L water
MgCl2: To make 1000ppm (1g/ L) Mg from MgCl2 Formula: Mg + 2(Cl) = x
Mg
-Y 24.305 + 2(35.453) = 3.9173 g/L 24.305
Dissolve 3.9173 g of MgCl2 in 1 L water
CaCl2: To make 1000ppm (1g/ L) Ca from CaCl2 Formula: Ca + 2(Cl) = x
Ca
-Y 40.078 + 2(35.453)  = 2.7692 g/L
40.078
Dissolve 2.7692 g of CaCl2 in 1 L water
KCl: To make 1000ppm (1g/ L) K from KCl Formula: K + Cl = x
K
103
-Y 39.098 + 35.453  = 1.907 g/L
39.098
Dissolve 1.907 g of KCl in 1 L water
104
Appendix ii
Ranki ng test for Preference Name:
Date:
Y ou are presented with four coded sampl es representi ng a range of four cereal bars.
Pl ease eval uate (accordi ng to appearance, aroma and taste) sampl e coded 102 fi rst, fol l owed by each of the other sampl es in the order of your choi ce. Pl ease ri nse wi th water between each tasti ng.
Write the coded sampl es in order of your preference in the spaces below. Appearance
1          2          3          4
Most    Least
preferred          preferred
Aroma
1          2          3          4
Most    Least
preferred          preferred
Taste

1          2          3          4
Most    Least
preferred          preferred
Comments:
105
Paired PreFerence Test Date:
Name:
Y ou wi ll receive two sets of cereal bar sampl es. Pl ease eval uate the sets (accordi ng to appearance), begi nning with the set nearest to you, maki ng your decision before proceedi ng to the next set.
C i rcl e in each set the sampl e you prefer.

Comments:
106
Appendix iii

Figure 1: Test plate for ranki ng test for preference.
Figure 2:Test plate for ranki ng test for preference.
107
Appendix iv
Figure 3:Calci um cal i bration curve. Cali bration curve representi ng calcium, resulti ng from data produced by the atomi c absorption spectrometer.
Figure 4: M agnesi um cal i brati on curve. Cal i brati on curve representi ng magnesi um, resul ti ng from data produced by the atomi c absorption spectrometer.
Figure 5: Potassium cal i brati on curve. Cal i brati on curve representi ng potassium, resulti ng from data produced by the atomi c absorption spectrometer.

Ca calibration curve
y = 0.0138x + 0.0042
R2 = 0.9967
Series1
Linear (Series1)
0          5          10        15
0.16
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
Mg calibration curve
-0.2 0  0.5       1          1.5
y = 0.9396x – 0.0487
R2 = 0.9895
Series1
Linear (Series1)

1 0.8 0.6 0.4 0.2 0

K calibration curve y = 0.2332x + 0.1503
R2 = 0.9283

2 1.5 1 0.5 0
	0          2          4          6          8

108
Appendix v
Figure 6: Standard curve of AA in water for D PPH .
Figure 7: Standard curve of AA in acetone/methanol for D PPH .

Figure 8: Standard curve of AA in water for FRA P.
0          20        40        60
y = 1.4924x – 3.5982
RZ = 0.9961
Series1
Linear …

80 60 40 20 0

0          20        40        60
RZ = 0.9865
Series1
Linear (Series1)
80
70
60
50
40
30
20
10
0
y = 1.6858x + 2.0498

0          20        40        60
Series1
Linear (Series1)
y = 0.0161x – 0.0402
RZ = 0.9902
1
0.8
0.6
0.4
0.2
0
109
Series1

0.7
		y = 0.0296x – 0.0343
0.6 0.5 0.4 0.3 0.2 0.1 0
			RZ = 0.9843

0          10        20        30
Figure 9: Standard curve of AA in acetone/methanol for FRA P.
110
Appendix vi
Table 1: H ydrophi l i c dilution cal cul ati ons. Cal cul ati ons used to determi ne the content of baobab (mg/mL) present in each hydrophi l i c dilution.

Table 2: L i pophilic dilution cal cul ati ons. Cal cul ati ons used to determi ne the content of baobab (mg/mL) present in each lipophilic dilution.

111
Appendix vi i
Table 3: Resul ts of pai red preference test

Where:
43 and 44 = no oats
43
112
Table 4: Resul ts of ranki ng test for preference-appearance

W here:
102= Plain
2
248= Coconut
3
912= Cocoa

2
319=C i nnamon
2
113
Table 5: Resul ts of ranki ng test for preference-aroma

W here: 3 102= Plain
1
248= Coconut
3
912= Cocoa
4
319=Cinnamon
2
114
Table 6: Results of ranking test for preference-flavour

W here: 102= Plain
1
248= Coconut
2
912= Cocoa
2
319=C i1nnamon

115
Appendix viii
Table 7: Cereal bar col our readi ngs, day 1 and day 8.

116

116
Appendix ix
Table 8: Calculations for nutritional information.

Baobab

Tags: Baobab health benefits, Baobab pulp Fruit Powder

Baobab Botanical Facial Masque

Discover the beauty secret of our innovative Baobab Botanical facial masque, an excellent source of trace minerals which can contribute to the strength of our connective tissue. The nutritional skin benefits include phyto-chemicals, enzymes and vitamins essential for strong cell growth and repair.

Tags: Baobab cosmetics, Baobab pulp Fruit Powder, Health benefits

The natural antioxidant properties of BAOMIX play an essential role in combating free radicals, the proliferation of which contributes to the premature aging of cells.  These antioxidants are integral to various metabolic processes, such as collagen production, the synthesis of hormones (steroids), and the production of connective tissue and neurotransmitters.
Ascorbic acid, also found in BAOMIX, improves the body’s ability to assimilate and distribute calcium and iron.
BAOMIX is especially recommended for anyone who desires to rebuild or maintain his or her good health:  seniors, children going through growth spurts, students, and athletes.  BAOMIX is a dietary supplement, and should not be used as a substitute for a varied and balanced diet.
100% organic and gluten free, BAOMIX is made by separating the naturally dried skin from the interior pulp of the fruit.
Directions:  Dissolve two teaspoons in a glass of water, fruit juice, iced tea, milk or yogurt one to two times daily.  Try it also as a breakfast tonic by adding two teaspoons to a cup of hot chocolate in the morning.  You can find more recipes at BAOMIX.COM.
Ingredients: 100% organic pulp of baobab (Adansonia digitata).  With a sweet and tangy taste, the pulp contains thiamine (vitamin B1) and riboflavin (vitamin B2), both essential to the regeneration of skin stem cells, and niacin (vitamin B3), which plays in important role in various metabolic functions.  Baobab pulp is naturally rich in minerals—calcium, iron, potassium, magnesium, manganese, phosphorus and zinc—and in several essential amino acids.
2 teaspoons of BAOMIX contain 44% of your required daily fiber, of which 22.4% is soluble and 22.6% insoluble.  Soluble fiber balances and fortifies your intestinal flora, which helps to facilitate healthy digestion.
An excellent complement to an active diet, 100 grams of BAOMIX contains seven times more vitamin C (300 mg) than an equal quantity of orange and three times more calcium (295 mg) than milk.
Store in a cool dry place.
Composition of 100 grams of baobab fruit:  75.6% carbohydrates, 2.3% protein, 0.27% lipids and 300 mg vitamin C.

Origin Senegal, distributed by company AGOJI France

Tags: antioxidant, Baobab pulp Fruit Powder, Health benefits, vitamin E

Madagascar is an island nation in the Indian Ocean, off the south eastern coast of Africa. The main island also called Madagascar which is the fourth largest island in the world and is home to five percent of the worlds plant and animal species. Most notable are the lemur of primates, the carnivorous fosse, three endemic bird families and six endemic baobab species.

Despite its location close to the African continent, the first human settlers of Madagascar appear to have come from Asia, rather than Africa. The culture shows the influence of both Africa and Asia.

The Malagasy language is of Malaya Polynesian origin and is generally spoken throughout the island. French is spoken among the educated population of this former French colony. English, although still rare, is becoming more widely spoken. Indians in Madagascar descend mostly from traders who arrived in the newly independent nation looking for better opportunities. The majority of them came from the west coast of India known as Muslim and Hindu. The majority speak Hindi or Gujarati, and though some other Indian dialects also exist a large number of the Indians in Madagascar have a high level of education, particularly the younger generation, which attempts to contribute their knowledge to the development of Madagascar.

Madagascar has got several microclimates due to the variation of the altitudes and its regional ecosystems. The seasons are mainly divided into two main periods. The rainy season from November to March and the dry season from April to October. The length of each period varies from one region to another one.

A holiday in Madagascar is unique, with its rich flora and fauna of such amazing variety and diversity that can be found nowhere else on earth. The warm and friendly people of Madagascar are African and Asian, proud to be from such a racially diverse, culturally rich country.

Madagascar gives rise to its astonishing biological diversity and remarkable scenery. Its of coastline is made up of mangrove reserves, stunning white sandy beaches, crystal clear waters, uninhabited land and coral reef. Madagascar has indeed been geographically blessed.

Madagascar is often referred to as the eighth continent, made up of endemic flora, fauna and wildlife species. It is most famous for its 50 species of lemurs these bright eyed mammals are often seen along with geckoes and chameleons.

Whale watching season is from July to September when humpback whales come into the St Marie channel to mate and give birth. Take part in the observation and preservation of humpback whales in the Indian Ocean.

It is incredibly unspoilt and offers adventure, beautiful beaches and deserts.

Article Source: http://www.ArticleStreet.com/

Tags: Baobab pulp Fruit Powder, Fruit pulp, Madagascar, organic

Baobab is a fruit from Africa that has recently been approved for use in the EU and UK. The baobab super-fruit is naturally dehydrated. It grows on the African baobab tree, and is encased in a thick hard shell. The fruit pulp itself is white, and is clumped around seeds and red fibers inside the shell.

The fruit pulp is mechanically removed from the shell and seperated from the red fiber. It has a nutty, acidic taste. Baobab pulp is only about 10% water, naturally, and contains significantly more vitamin C than oranges. The dehydrated form of the pulp means that the fruit does not need to be processed, just mechanically seperate, and use. In the EU and UK, it is intended to use baobab fruit pulp for smothies and cereal bars, amongst other possible uses.

Tags: Baobab health benefits, Baobab pulp Fruit Powder

The first natural bio complex created, dehydrated and stabilized by nature

Adansonia Digita, or Baobab, bears fruits once a year, which are then harvested by the local community. The Fruit pods contain seeds, red funicles and naturally dehydrated fruit pulp.
The Fruit pulp is a natural, non treated organic raw material, which is seperated from the seeds and the fibers by a mechanical process.

Why use Baobab pulp Fruit Powder

10 x the antioxidant level of oranges
6 x more vitamin C than oranges
6 x more antioxidants than Cranberries, Blueberries, and Blackberries
6 x Times More Potassium than a Banana
2 x More antioxidants than Goji Berries
Omega 3, 6, & 9
More Iron than red meat
More Magnesium than Spinach
Twice the calcium level of milk
Valuable aid in the prevention and treatment of gastric and IBS disorders
Effective in treating osteoporosis, varicose veins and hemorrhoids
The fruit’s minerals and vitamins are beneficial to anaemics, anorexics, smokers and athletes
Excellent ingredient in diets for diabetics (notably type II diabetes)
Displays antioxidant capacity by fighting against the formation of free radicals
Ideal in pre-biotic formulations, and stimulates the intestinal micro flora
Great complement in diets for celiac diseases
Involved in Tempe fermentation
Antiviral properties
Energy Booster
Diet Supplement
Baobab Fruit powder can be added to energy drinks, chewable tablets and vitamin bars to aid a diet supplement
Functional Food
Baobab fruit powder can be added to Yoghurts (with pre or pro biotic activities), biscuits, ice cream, fruit juice, smoothies and gluten free products
Daily Supplement
Can be added to cereal and powdered shakes as part of a daily food

Independent tests have also confirmed Baobab Fruit Powder’s importance as an anti-inflammatory, antibacterial, antifungal, antipyretic and analgesic agent.

Nutritional information per 100g

* Energetic Value (kcal)- 175 Kcal/100g
* Energetic Value (KJ) – 750 Kj/100g
* Proteins – 2.9g/100g
* Fats – 0.4g/100g
* Total Carbohydrates – 39.0g/100g
* Dietary Fiber – 45.9g/100g
* Sodium – 0.2mg/100g

Proteins
Average values of aminoacids per 100g of protein

* Proline – 2.35 g
* Histidine – 2.71 g
* Leucine – 8.41 mg
* Lysine – 14.62 g
* Arginine – 6.04 g
* Isoleucine – 10.73 g
* Methionine – 4.92 g
* Cysteine – 11.23 g
* Glutamic acid – 4.02 g
* Valine – 1.62 g
* Tyrosine – 4.21 g
* Tryptophan – 1.49 g
* Threonine – 2.96 g

Dietary Fibers
Average values per 100g of fruit pulp.

* Soluble dietary fibers – 23g/100g
* Insoluble dietary fibers – 23g/100g

Sugars
Average values per 100g of fruit pulp.

* Gluctose – 3.4g/100g
* Fructose – 3.3g/100g
* Saccharose – 20g/100g

Minerals
Average values per 100g of fruit pulp.

* Calcium – 300mg/100g
* Phosphorus – 96-210mg/Kg
* Iron – 2.7mg/Kg
* Potassium – 2.31mg/Kg
* Manganese – 0.60-0.90mg/Kg

Vitamins

* Vitamin C – 295mg/100g
* Vitamin A – 200mcg/100g
* Vitamin B1(thiamin) – 0.04mg/100g
* Vitamin B2(riboflavin) – 0.06mg/100g
* Vitamin B6(piridoxin) – 2.13mg/100g
* Vitamin PP(niacin) – 2.16mg/100g

sources : www.baobabfruit.co.uk

Tags: Baobab health benefits, Baobab pulp Fruit Powder

バオバブ

バオバブ（英名：Baobab、学名：Adansonia）はアオイ目アオイ科（クロンキスト体系や新エングラー体系ではパンヤ科）バオバブ属の総称のこと。

学名はA. digitataを報告した仏人自然学者Michel Adansonの名に由来する。原生種がマダガスカルに6種、オーストラリアとアフリカに1種ずつ存在する。
サバンナ地帯に多く分布する。幹は徳利のような形をしており、高さは約20メートル、直径は約10メートルに及ぶ。最大のものは南アフリカのリンポポにあ る高さ47m、直径15mである。年輪が無いため樹齢を知ることは難しいが、数千年に達すると言われ、放射性年代測定は可能である。中は空洞になることが 多い。葉は幹の上部につき、乾季に落葉する。花は白色で大きい。果実はヘチマのように垂れ下がり、堅い。果肉は食用・調味料とされ、セネガルでは「サルの パン」と呼ばれる。ビタミンCがオレンジより多く、カルシウムも牛乳より多いと言われる。また、若葉を野菜として利用する。樹皮は煎じて解熱剤に用いられ る[1]。

その独特の樹型から、悪魔が大木を引き抜いて、逆さまに突っ込んだといわれている。 サン・テグジュペリの『星の王子さま』では、星を破壊する巨木として描かれている。浜名湖花博において日本では初めて屋外で開花した。

120klもの水分を幹にたくわえており、乾季になると葉を落とし休眠する。休眠中はその水分で生きのびる。 バオバブは観葉植物にもなり、盆栽型に仕立てることもできる。

また、このタネは、オランウータンに一度食べられ排泄された物でないと発芽しないと言われていた。

japan japon 日本

Tags: Baobab pulp Fruit Powder, Botanic of the Baobab tree, Health benefits, バオバブ