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Continental J. Food Science and Technology 2: 1 - 5, 2008 © Wilolud Online Journals, 2008.

EFFECTS OF ROXARSONE ON COPPER UTILIZATION BY CHICK

Gabriel O. Wordu and P. Obinna -Echem Department of Food Science & Technology, Faculty of Agriculture, Rivers State University of Science &

Technology, P.M.B. 5080, Port Harcourt, Rivers State, Nigeria

ABSTRACT Experiment was conducted to examine the effect of dietary roxarsone (3-nitro-4-hydroxyphemylarsonic acid) on Cu utilization by chick. A fortified corn-soybean meal diet was fed to the chick. Roxarsone dramatically reduced liver cu concentration at all levels of supplemental Cu fed. The level of roxarsone commonly fed, 50 mg/kg diet, resulted in a two-to-four fold depression in liver Cu concentration in chick. The effects of roxarsone on weight gain were more perplexity. In the chick, the diets containing 100 and 250 mg Cu/kg depressed growth in the presence, but not in the absence, of 50mg/kg dietary roxarsone. In contrast, at toxic levels of Cu, roxarsone had no effect on (500 or 750 mg Cu/kg diet) the growth-depressing effects of Cu/kg diet) the growth-depressing effects of Cu. KEY WORDS: Roxarsone, Copper, Chick, Liver Copper, Toxicity

INTRODUCTION Roxarsone, (3-nitro-4-hydroxyphonylarsonic acid) figure 1, is commonly used a feed additive in both poultry and susine feeds. Although the toxic effects of roxarsone have been studied (Wise et al; 1994; Rice et al; 1980), the interactions between roxarsone and another common feed additive, Cu, have not been extensively studied, using chick as model. It has been reported that roxarsone dramatically reduces liver Cu concentration in chicks fed supplemental Cu (czarnecki and Bakar, 1984). In addition, a growth depression is generally observed when growth-promoting levels of roxarsone and Cu are combined in the diet. This study was to examine the Cu and roxarsone interaction and to determine whether it is unique for the poultry and the utilization of cu as affected by roxarsone. MATERIALS AND METHODS The basal diet is presented in table 1. This diet was designed to meet nutrient requirement of the chick. Supplemental roxarsone was supplied as a 10% premic and Cu as CuSo4 .5H2O. All dietary additions were made at the expense of corn-starch. Feed and water were provided addibitum throughout all experiments. Broiler-type chicks were used in the experiment. The chicks were fed a corn-soybean meal starter diet during the first 7 d posthatching. After an overnight fast, experimental groups of five male chicks were selected to have similar mean initial weight and weight distributions. Triplicate groups of five male chicks were assigned to each treatment. Assay length was 8 to 23 d post hatching and averaged initial weight was 81.7g. The chicks were fed 0, 100, 250, 500, 750 or 1000 mg supplemental Cu/kg diet in the presence or absence of 50mg/kg supplemental roxarsone constituting a 6 x 2 factorial treatment design. Fifty milligrams/kilogram roxarsone and between 100 and 250 mg/kg Cu are the common use levels in commercial boiler production.

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G. O. Wordu and P. Obinna –Echem: Continental J. Food Science and Technology 2: 1 - 5, 2008 At the termination of the experiment, liver and bile sample were obtained per replicate. Tissues samples were dried at 100C for 24h, weighed, wet ashed with HNO3 and analysed for Cu by atomic absorption spectrophometry (AOAC, 1984). Results are expressed as gµ cu/g dry tissue.

Data were analyzed by analysis of variance, using methods appropriate for factorial treatment designs (Steel and Torrie, 1980). RESULTS AND DISCUSSION The results of the experiment are presented in figure 2 and table 2. Low levels of Cu (100 and 250mg Cu/kg diet) depressed growth in the presence, but not in the absence, of 50 mg/kg dietary roxarsone. This resulted in a roxarsone (O vs 50) x Cu (0 vs 100, 250) interaction (p<.05). In contrast, roxarsone slightly ameliorated (1<.06) the toxicity of 1,000mg Cu/kg diet, while it had had effect on the growth-depressing effect of 500 or 750mg Cu/kg diet. Liver Cu concentration was dramatically reduced by roxarsone at all levels of supplemental Cu addition (P<.05). Bile Cu was similarly affected. The depression in liver Cu concentration due to feeding roxarsone was not specific for the avian species. In fact, 50mg/kg roxarsone resulted in approximately the same magnitude depression in liver Cu concentration in pig and rat (Czarnechi and Baker 1983) The roxarsone probably reduced liver Cu concentration by inhibiting Cu absorption rather than by enhancing Cu excretion. Although, roxarsone consistently decreased liver Cu concentration at all levels of supplemental Cu intake, the effects of roxarsone on performance can be obtained from either roxarsone (50mg/kg) or Cu (100 to 250mg/kg) When fed alone, the combination produced a growth depression in chick (figure 2). Similar results were reported by Czarnechi and Baker (1984). This interaction may have important practical implications for the broiler chick. However, the combination of growth-promoting levels of roxarsone and Cu does not produce growth in pigs (Czarnecki and Baker, 1984). It is interesting and somewhat perplexing that roxarsone can either increase, decrease or have no effect on weight gain of chick fed supplemental Cu, depending on the level of Cu fed. Nonetheless, it consistently reduced liver Cu concentration. Table 1. Composition of chick Basal Diet

Ingredient % Cornstarch 100 Corn meal 55.10 Soybean meal 25.08 Corn glufen meal 10.00 Corn oil 5.00 Dicalcium phosphate 2.20 Ground limestone 1.00 Iodized salt .40 L-lysine-HCL .25 L-Methionine .20 Vitamin premix .10 Choline.Cl .10 Ferric citrate .06 MnSO4 .05 ZnCO3 .01

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G. O. Wordu and P. Obinna –Echem: Continental J. Food Science and Technology 2: 1 - 5, 2008 Figure 1: Structure of roxarsone (3-nitro-4-hydro-xyphenylarsonic acid) Table 2. Tissue copper concentration of Chicks fed copper with or without supplemental Roxarsonea Liver Copperc Bile Copperc Copperb ( µ g/g dry tissue) (µ g/g dry bile) Mg/kg Roxarsone, mg/kg Roxarsone, mg/kg 0 50 0 50 0 15.6 14.8 21.1 24.0 100 39.0 17.9 31.7 29.4 250 71.8 18.2 54.9 43.0 500 531.1 138.8 143.6 112.5 750 960.8 449.4 197.8 161.6 1,000 2,184.6 894.6 345.6 173.1 aTissue samples from five birds within each replicate were pooled before atomic absorption analysis was performed. Pooled SE for the transformed data were .20 and .16 for liver and bile Cu, respectively. bprovided as CuSO4 :5H2O, cRoxarsone xCu interaction (p<.05).

ASo(OH)

NO2

OH

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G. O. Wordu and P. Obinna –Echem: Continental J. Food Science and Technology 2: 1 - 5, 2008

Dietary Cu (mg/kg diet)

Figure 2. Weight gain of Chicks fed graded levels of copper with or without 50mg supplemental roxarsone/kg diet. REFERENCES AOAC, (1984). Official Methods of Analysis (14th Ed.) Association of Official Analytical Chemists,

Washington, DC. Czarnecki, G. L; and Baker, D. H; (1984). Feed additive interactions in the chicken: Reduction of tissue

copper deposition by dietary roxarsone in healthy and in Eimerin acevulwia-or Eimeria Tenella-infected chicks. Poul. Sci. 63:1417.

Czarnecki, G. L; and Baker, D.H. (1983). The role of roxarsone as a metal chelating agent. Poul. Sci.

62:1407. Rice, D. A; McMurray, C. H; McGracken, R. M; Bryson, D. G. and Maybin, R. (1980) A field case of

poisoning caused by 3-nitro-4-hydroxyphenylarsonic acid in pigs. Vet. Res. 106:312. Steel, R. G. D. and Torrie, J. H. (1980) Principles and procedures of statistics: A biometrical Approach (2nd

Ed). McGraw-Hill Book Co., New York. Wise, D. R; Hartley, W. J. and Fowler, N. G; (1974). The pathology of 3-nitro-4-hydroxyphenylar sonic

acid toxity in Turkeys Res. Vet. Sci. 16:336

200

250

300

Gain

(g/

)

0 500 1000

Roxarsone

Basal diet

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G. O. Wordu and P. Obinna –Echem: Continental J. Food Science and Technology 2: 1 - 5, 2008 Received for Publication: 07/11/2007 Accepted for Publication: 22/12/2007 Corresponding Author: Gabriel O. Wordu Department Of Food Science & Technology, Faculty Of Agriculture, Rivers State University Of Science & Technology, P.M.B. 5080, Port Harcourt, Rivers State, Nigeria E-mail: wordugab@yahoo.com

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Continental J. Food Science and Technology 2: 6 - 13, 2008 © Wilolud Online Journals, 2008.

PRODUCTION, PROXIMATE COMPOSITION AND CONSUMER ACCEPTABILITY OF BISCUITS

FROM WHEAT/SOYBEAN FLOUR BLENDS

Okoye, J.I1, Nkwocha, A.C1 and Ogbonnaya, A.E.2 1Department of Food Science and Technology, 2Department of Biochemistry, Madonna University, Elele

Campus, P.M.B 48 Elele, Riversstate, Nigeria.

ABSTRACT The use of wheat and soybean flour blends in the preparation of biscuits was studied. The flour blends of wheat (WF) and soybean (SF) were composite at replacement levels of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80% and 90%. The proximate composition of the various flour blends used for the preparation of biscuits were determined using standard methods. The biscuits produced were evaluated for their proximate composition and sensory attributes. From the results, the protein content of the blends increased steadily with increasing content of soybean flour (SF) (13.18% in 90:10, WF: SF to 27.44% in 10: 90, WF: SF) while carbohydrate decreased. Similarly, the protein content of the biscuits increased with increasing supplementation with soybean flour. The protein content of biscuits increased from 12.08% in 90:10, WF: SF to 25.48% in 10: 90, WF: SF samples. In the same way, the energy content of the biscuits increased as the level of soybean flour inclusion increased. The energy content of the biscuits increased from 375.6kcal in 90: 10, WF: SF to 435.5kcal in 10: 90, WF: SF. The results also showed that the biscuits fortified with 10% soybean flour was the most acceptable because there was no significant difference (p>0.05) between this sample and the control (Biscuits made from 100% wheat flour). The other samples of biscuits were significantly different (p<0.05) from the control with the biscuits made from 100% soybean flour having the lowest score (4.8) in general acceptability. KEYWORDS: Biscuits, proximate composition, acceptability, wheat-soybean flour blends.

INTRODUCTION Biscuits are flour confections produced from dough or batter and baked to a very low moisture content within a short period of time to make them flaky and crispy. The consumption of biscuits and other western styled bakery products such as bread and cakes prepared from wheat flour has become very popular in Nigeria, especially, among children (Ayo and Nkama, 2003). The low protein content of wheat flour, which is the most important ingredient used for the production of different kinds of baked goods has been of major concern in its utilization. The enrichment or fortification of biscuits and other bakery products with other protein sources such as oilseeds and legumes has received considerable attention. This is because oil seed and legume proteins are high in lysine, an essential limiting amino acid in most cereals (Ihekoronye and Ngoddy, 1985). Whole legumes contain relatively high amount of protein compared to other plant foodstuffs. Legume proteins should complement the protein in cereal grains since the chemical and nutritional characteristics of legumes make them natural complements to cereal-based diets (Altschull, 1974). The soybean, (Glycine max) a grain legume, is one of the richest and cheapest sources of plant protein that can be used to improve the diet of millions of people, especially the poor and low income earners in

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 developing countries because it produces the greatest amount of protein used as food by man (LIU, 2000). Soybean can be processed into soymilk, soy sauce, tofu (soybean curd), soy-yoghurt, soy sprouts, soy flour and many other soy products. Defatted soybean flour can be used for the production of protein isolates and concentrates. Soybean seeds ↓ Washing ↓ Soaking in water (for 8h) ↓ Dehulling ↓

Draining and Removal of hulls ↓ Boiling (1000Cfor 30 min) ↓ Drying (650C for 6 hr) ↓ Milling (Attrition mill) ↓ Sieving (Fine sieve, 400µm) ↓ Cooked full-fat soybean flour ↓ Packaging

Fig 1: Flow Chart for The Production of Cooked full-fat Soybean flour.

Nutritionally, soybean protein resembles animal protein more closely than other vegetable proteins. Soybean protein constitutes about 40% of the total solids and plays a very important role in the enrichment of cereal-based baked goods (Fukushima, 1999). It is also a rich source of vitamin, minerals and is relatively low in crude fibre (Oyenuga, 1968). Soybean is one such protein sources, which when used partially to replace or complement wheat flour in the production of bakery products such as biscuits, bread and other confectionery could go a long way in improving the nutritional status of such products. The objective of the study was to determine the nutrient constituents and acceptability of biscuits prepared from wheat flour supplemented with soybean flour at different levels of substitution. MATERIALS AND METHODS Source of Materials Mature Soybean seeds (Glycine max) were bought from a local market in Owerri, Nigeria whereas the wheat flour and other ingredients used for this study were purchased at Ogbuke market, Elele, Nigeria. This research work was carried out in Department of Food Science and Technology, Madonna University, Elele, Rivers State, Nigeria. ( June, 2006) Preparation of Full-Fat Soybean Flour The full-fat soybean flour was prepared according to the method described by Ihekoronye and Ngoddy (1985) as shown in Figure 1. During preparation, two kilograms of soybean seeds which were free from dirts and other foreign particles such as stones, sticks and leaves were weighed, cleaned and soaked in tap water for 8 hours. Thereafter, the seeds were drained, dehulled manually, boiled (1000C, 30min) and dried

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 in cabinet dryer (650C, 6hr). During drying, the dehulled seeds were stirred at intervals of 30 minutes to ensure uniform drying. The dried seeds were milled (attrition mill) and sieved to obtain cooked full-fat soy bean flour. The full-fat soybean flour obtained was finally packaged in an air tight container due to the hygroscopic nature of soybean flour unit used for blending and analysis. Flour Blending Wheat flour (WF) was composite at 10, 20, 30 40, 50, 60, 70, 80, and 90% levels with soybean flour (SF) on a replacement basis in Kenwood blender. The flour blends were individually packaged in sealed polyethylene bags and kept at room temperature until used for biscuit production and analysis. The various flour blends are shown in Table 1

Ingredients

Mixing and aeration

Sheeting/rolling (on a wooden board with rolling pin)

Cutting (using biscuit cutter)

Printing

Baking (in a hot oven at 2000C for 20min)

Cooling

Scraps separation

Weighing

Packaging

Fig 2: Flow Chart for the Production of Biscuits

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 Preparation of Biscuits The biscuits were prepared using the creaming method described by Okaka (1997) as shown in Figure 2. The basic formulation was 100% flour, 40% fat, 20% beaten egg, 60% sucrose, 4% milk, 2% salt and 1% baking powder. The 100% flour was systematically replaced for the wheat flour (WF) with 10, 20, 30, 40, 50, 60, 70, 80, and 90% soybean flour (SF) During the biscuit making, the sugar and fat were initially creamed in a mixer (Model AT 220A) to produce a creamy mixture before the flour and other dry ingredients were added. Thereafter, the mixture was Table 1: Flour Blends

Samples WF (%) SF (%)

A 100 0

B 90 10

C 80 20

D 70 30

E 60 40

F 50 50

G 40 60

H 30 70

I 20 80

J 10 90

K 0 100

thoroughly mixed to form hard consistent dough. The dough obtained was thoroughly kneaded manually on a smooth clean table for about 5 mins. The dough was thinly rolled on a wooden board with rolling pin to uniform thickness (2mm) and cut out (using biscuit cutter) to desired shapes of similar sizes. The cut out biscuit dough pieces were placed in a greased baking tray and baked in a hot oven (2000C) for 20 mins. The biscuits were cooled immediately after baking and packaged individually in an air tight container and kept at room temperature until used for analysis and sensory evaluation. Wheat biscuits were similarly baked/produced as reference. Chemical Evaluation Proximate analysis was carried out on each of the flour blends and the biscuits. All determinations were carried out in triplicates. Protein determination was by kjeldahl method (Pearson, 1973). Fat, ash and moisture content determination methods were as described by Pearson (1973). The carbohydrate was determined by difference (Bryant et al, 1988). Food energy was calculated using the Atwater factor 4 x protein, 4 x carbohydrate, 9 x fat (Marero et al, 1988).

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 Sensory Evaluation The biscuits were evaluated by a panel of fifteen untrained judge drawn from the University Community for attributes of colour, texture, flavour and general acceptability on a hedonic scale of 1-9 where 1 = dislike extremely and 9 = like extremely (Ihekoronye and Ngoddy 1985). Statistical Analysis Analysis of variance (ANOVA) was used in all the analysis for detection of significant differences (p<0.05) among samples. The turkey test was used in separating significant means. RESULTS AND DISCUSSION The proximate composition of the various flour blends are shown in Table 2. The moisture content ranged from 12.27% to 14.62%. The differences could be attributed to inadequate drying of soybean seeds after soaking and boiling. The wheat flour (WF) appeared to be better dried as reflected in the moisture content values for the flour. The ash content of all the flour blends differed significantly (p<0.05) from each other. The differences were observed because the ash content of the blends increased steadily with increasing content of soybean flour (SF). Legumes have been reported to be good sources of ash (Pyke, 1981). This addition effect was also observed for fat and protein. In other words, the ash, fat and protein contents of the blends increased as the level of soybean flour inclusion increased. However, the carbohydrate content of the WF/SF blends decreased with the increasing concentration of soybean flour (SF). The result showed that soybeans are not good sources of carbohydrate when compared to other legumes (Salunkhe et al, 1992). Table 2: Means 1, 2 of Proximate Composition of Wheat/Soybean flour blends on dry weight basis.

Samples Moisture (%)

Ash (%) Fat (%) Nx5.75 Protein (%)

Carbohydrate (%)

A 12.27a 0.86a 0.82a 11.62a 73.43a B 12.42a 1.88b 3.22b 13.18b 69.30b C 12.64a 2.08c 4.68b 14.46ab 65.92b D 13.28b 2.42c 6.28c 16.32c 62.80b E 13.26a 2.46c 7.86c 16.86c 59.57c F 11.62c 2.86c 8.42ab 17.44d 60.66c G 12.82a 3.18d 9.64d 19.22ac 57.14ab H 12.22a 3.32d 12.18e 22.82e 51.66ac I 12.62a 3.56a 14.08f 24.24de 46.50d J 13.42b 3.86d 16.02ac 27.44f 42.85dc K 14.62d 4.62ab 18.46de 25.64fe 34.86e 1 Means of triplicate samples 2 Means in the same column and followed by the same letters are not significantly different from

each other (p>0.05) The supplementation of wheat flour (WF) with soybean flour (SF) produced the desired effect of increasing the protein content of the blends, which will in turn improve the nutritional quality of biscuits produced from these blends. The proximate composition of biscuits made from WF/SF blends are shown in Table 3. The moisture content of all the biscuit samples was not significantly different from each other (p>0.05). They were also similar with those reported by Akpapunam and Darbe (1994). The protein contents was significantly different (P<0.05). The biscuits produced from blends with higher concentrations of soybean

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 flour contained more protein than the ones made from blends containing less proportion of soybean flour. There was a gradual decrease in the carbohydrate content of the biscuits from sample A to sample K. The ash content of all the samples was higher than those previously reported for biscuits. The fat content of the biscuit samples was similarly higher than those reported earlier for biscuits. The increase in fat content of all the biscuit samples could be attributed to the fact that the soybean flour was not defatted. However, soybeans have been reported to be good sources of fat (Iwe and Onuh, 1992). The energy content of all the biscuit samples ranged from 366.28 kcal/ 100g to 435.48 kcal/100g. There was significant difference in the energy content of the biscuits (p<0.05). The energy contents of the biscuit samples were higher than those reported by Oyewole et al, (1996). The increase in energy content of the biscuits resulted from the high protein, fat and carbohydrate contents of the blends used for their production. Table 3: Means 1, 2 of Proximate Composition of Biscuits Produced from WF, SF and WF:SF Blends

Samples Moisture (%)

Ash (%) Fat (%) Nx5.75 Protein (%)

Carbohydrate (%)

Energy (Kcal/100g)

A 9.86a 1.42a 2.28a 11.22a 75.22a 366.28a B 9.44a 1.76b 4.08b 12.08a 72.64a 375.60a C 9.62a 1.82b 6.22c 14.56b 67.74b 385.28a D 9.68a 2.02c 8.26d 15.12b 64.74b 385.28a E 9.86aa 2.12c 9.82e 16.64c 62.17c 401.14c F 9.48a 2.18c 11.32ab 17.68c 59.43d 409.96c G 9.28a 2.28c 13.28ac 18.08ab 58.08d 424.26d H 9.42a 2.44c 14.48f 20.28d 54.48ab 428.96d I 9.62a 3.62d 16.22bc 22.46e 49.08e 432.24d J 9.74a 3.86ab 16.88bc 25.48de 47.56ac 435.48e K 9.36a 3.74d 17.22de 24.28de 44.58de 432.22e 1 Means of triplicate samples 2 Means in the same column and followed by the same letters are not significantly different from

each other (p>0.05) The scores of the flair sensory evaluation of the biscuits produced from wheat/soybean flour blends are shown in Table 4. The scores of various sensory attributes were low in all the biscuits samples from the different blends. In general, biscuits produced from 100% wheat flour (Sample A) used as control were better accepted by the judges and were not significantly different from each other. However, the unique baking property of wheat flour has been well known (Ihekoronye and Ngoddy, 1985). Wheat flour generally has a better baking quality than any other type of flour. The substitution of wheat flour (WF) with soybean flour (SF) in all the biscuit samples up to 60% produced good results. Beyond this level, the sensory scores were very poor on the average. Acceptable results were also reported for African bread fruit kernel / wheat flour blended biscuits (Akubor et al, 2000). CONCLUSION Biscuits of acceptable quality similar to those made from wheat flour were produced from WF/SF blends. Substitution of wheat flour (WF) with soybean flour (SF) beyond 60% did not produce good results. Therefore, substitution beyond this level is not encouraged. From the study, it was observed that the biscuits produced had better nutritional quality than those produced from 100% wheat flour because of their high protein content despite the high level of sugar (Sucrose) used for the production of biscuits which may impair protein availability due to the incidence of maillard or carbonyl amine reaction. Also, further studies

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 should be performed on wheat flour/soybean blended biscuits to evaluate their respective protein quality and availability. The enhancement of the nutritional value of biscuits with the addition of soybean flour, could help to alleviate the problem of protein – energy malnutrition prevalent in Nigeria and other developing countries of the tropics Table 4: Means 1, 2 of Sensory evaluation of Biscuits Produced from WF, SF and WF:SF Blends.

Sample Colour Texture Flavour General Acceptability

A 4.8c 6.4a 7.2c 7.8a B 5.9b 6.1a 6.7a 7.4a C 6.4a 5.8b 6.4a 6.6b D 5.2b 6.2a 6.3a 6.4b E 5.6b 5.6b 5.8b 6.2b F 6.0a 5.4b 5.6b 5.7c G 5.4b 4.5c 5.4b 5.6c H 5.6a 4.2c 4.8c 5.8c I 5.8b 4.6c 4.6c 5.6c J 6.2a 5.2ab 4.4c 5.4c K 6.8a 4.7c 4.2c 4.8d 1 Means of 15 untrained Panelists / Judges 2 Means in the same column and followed by the same letters are not significantly different from

each other (P>0.05) ACKNOWLEDGEMENT The authors thank Messers E.O Agbonwaneten and E.O. Ozochi for their invaluable contribution to this study. Moral support received from Mr. E.E Ononobi of Department of Food Science and Technology, Madonna University, Elele Campus, Rivers State, is highly appreciated. REFERENCES Akpapunam, M.A and Darbe, J.W. (1994). Chemical composition and Functional Properties of Maize and bambara groundnut flours for cookie production. Plant foods. Hum Nut 46: 147-155 Akubor, P.I., Isolokwu, P.C., Ugbane, O. and Onimawo, I.A (2000). Proximate Composition and functional properties of African bread fruit kernel and wheat flour blends. Food Res. Inter. 33:707-712. Altschull, A.M. (1994). New protein foods. Vol. 1a. Academic Press Ltd, Ibadan. pp 20-24 Ayo, J.A and Nkama, I (2003). Effect of acha (D.exilis) flour on the physico chemical and sensory qualities of biscuits. Nutrition and food science 33(3): 125-130. Byrant, L.A; Montecalvo, J.J.R; Morey, K.S. and Lay, B. (1988). Processing, functional and nutritional properties of okro seed products. Journal of Food Science 53: 810-816. Fukushima, D. (1999). Recent progress of soybean protein foods: Chemistry, Technology and Nutrition. Food Review Int. 7(B): 323-352. Ihekoronye, A.I and Ngoddy, P.O (1985). Integrated Food Science and Technology for the Tropics. Macmillian Publishers Ltd, London and Oxford. Pp 283-292

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Okoye, J.I et al: Continental J. Food Science and Technology 2: 6 - 13, 2008 Iwe, M.O. and Onuh, J.O. (1992). Functional properties of soybean and sweet potato flour mixtures. Lebensin – Wiss. U-Technol. 25: 569-573 LIU, K. (2000). Expanding soybean food utilization. J. Food Technol. 54(7): 46-47. Marero, L.M., Payuma, E.M., Librando, E.C., Lainez, W.N., Gopz, M.D. and Homma, S. (1988). Technology of Weaning food formulations prepared from germinated cereals and legumes. Journal of Food Science 53(5): 1391 – 1395. Okaka, J.C. (1997). Biscuit Manufacture. In cereals and legumes storage and processing Technology. Data and Microsystems Publishers Ltd Enugu. pp 115-130. Oyenuga, V.A. (1968|). Nigeria’s food and feeding stuffs, their chemistry and nutritive value 3rd edition, Reprinted, Ibadan University Press, pp 20-26. Oyewole, O.B., Sannic, L.O. and Ogunjobi, M.A. (1996). Production of biscuits using cassava flour. Nigeria Food Journal 14: 24-29 Pearson, D. (1973). Laboratory Techniques in Food Analysis. Butterworth and Company Publishers Ltd, London, pp 27-72. Pyke, M. (1981). Classification of Wheat: Food Science and Technology. 4th edition. John Murray Publishers Ltd, London, pp 44-56. Salunkhe, D.K., Charan, J.C., Adesule, R.N., and Kadam, S.S (1992). World oil seed, chemistry, Technology and Utilization. An Avi Book Published by Van Nostrand Reinhold, New York, pp 115-136. Received for Publication: 20/10/2007 Accepted for Publication: 22/12/2007 Corresponding Author: Nkwocha, A.C Department of Food Science and Technology, Madonna University, Elele Campus, P.M.B 48 Elele, Rivers State, Nigeria. E-mail: Chinkwocha1@yahoo.com.

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Continental J. Food Science and Technology 2: 14 – 19, 2008 © Wilolud Online Journals, 2008.

EFFECT OF MALTING AND SOYBEAN SUPPLEMENTATION ON THE NUTRIENT QUALITY AND ACCEPTABILITY OF “EKO-EDA”: A MAIZE GRIT PORRIDGE

OCHEME, O.B1.; ALASH, A.M.1 and ZAKARI, U.M2. 1Department of Food Science and Nutrition, Federal University of Technology, P.M.B. 65, Minna, Niger

State, Nigeria. 2Department of Food Science and Technology, Federal Polytechnic, Kaduna, Kaduna State, Nigeria.

ABSTRACT The effect of malting and soybean supplementation on the nutrient quality and acceptability of maize grit porridge was investigated. Malting of maize and supplementation with soybean significantly (P<0.05) increased protein, ash and copper content of eko-eda while fat and crude fibre were significantly (P<0.05) decreased. Soybean supplementation significantly (P<0.05) increased iron and phosphorus content. With the exception of methionine and tryptophan, all essential amino acids were adequate. Maize grit porridge (eko-eda) prepared from malted maize grits and blends of maize/soybean grits were acceptable.

KEYWORDS: Malting, supplementation, porridge, essential amino acids, nutrient, quality.

INTORDUCTION Maize (Zea mays), the American-Indian word for corn, literally means, “that which sustains life” (FAO, 1997). It is, after wheat and rice, the most important cereal grain in the world providing nutrients for humans and animals and serving as a basic raw material for the production of starch, oil, alcoholic beverages, food sweeteners and more recently, fuel. Maize is an important food in Asia, Africa, Latin America and part of Europe. Each country in these continents has one or more maize dish(es) unique to its culture. Examples are “kenkey” in Ghana; koga in Cameroun; injera in Ethiopia; and ugali in Kenya just to mention a few. Likewise in Nigeria, each locality or culture has one or more maize dish(es) unique to its people. An example is “eko-eda”. Eko-eda, a maize grit porridge, is a popular breakfast meal of the Yoruba-Igbomina tribe, natives of Ifelodun and Irepodun Local Government areas of Kwara State in central Nigeria. It is produced by dry milling maize grains into grit and sieving into various particle sizes, any of which can be used depending on individual preferences. The grits are then soaked in water and the hulls and germs removed by decanting. This process gives a cleaner and more acceptable grit which is then steeped in water overnight to soften. The soft maize grits are then cooked in boiling water and allowed to thicken to an acceptable consistency, this also depends on individual preferences. It is then served hot. Sugar (sucrose) may be added to enhance the taste. Maize constitutes an important source of carbohydrates, protein, B-complex vitamins and minerals. As an energy source, it compares favourably with root/tuber crops and is similar in energy value to dried legumes. Furthermore, it is an excellent source of carbohydrate and is richer in nutrients compared to other cereals. However, maize is deficient in lysine and tryptophan. Many studies conducted with animals have demonstrated that the addition of both amino acids improves the quality of maize protein. The addition of these limiting amino acids has been confirmed to improve the nitrogen balance in children (Bressani et. al., 1968).

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OCHEME, O.B et al: Continental J. Food Science and Technology 2: 14 – 19, 2008 Soybeans (Glycine max) is rich in both essential and non-essential amino acids. For a plant food, soybean protein is exceptionally rich in lysine and can serve as a valuable supplement to cereal foods where lysine is a limiting factor (FAO/WHO, 1973). Though a valuable source of protein, soybean has certain adverse nutritional effects following consumption of raw soybean meal. This has been attributed to the presence of endogenous inhibitors of digestive enzymes. The full nutritional potential of soybean is attained only after a certain amount of heat has been applied. Table 1: Chemical composition of maize grits and blends of maize-soybean grits.

Grit samples Parameter C MMG MMSG MMMSG Moisture (%) 9.43b 11.76a 8.44d 8.98c Protein (%) 8.57c 6.89d 17.25b 20.26a Fat (%) 12.38a 10.70a 8.19b 5.49c Ash (%) 0.33c 0.45b 0.67a 0.68a Crude fibre (%) 4.05a 2.38b 2.06b 1.27c Carbohydrate (%) 65.24b 67.82a 63.39c 63.32c Calcium (mg/kg) 96.2a 96.4a 96.3a 96.0a Copper (mg/kg) 4.71c 4.86b 5.26a 5.28a Iron (mg/kg) 3.4b 3.4b 5.47a 5.49a Phosphorus (mg/kg) 442.5b 451.1b 563.7a 5573b

Values are means of triplicate determinations, Means in the same row with different superscript are significantly different (p<0.05) C = Untreated maize grits (control) MMG = Malted maize grits

MMSG = 70:30 blend of untreated maize grits and malted soybean grits MMMSG = 70:30 blend of malted maize and malted soybean grits.

Malting involves the germination of grains until the food store (endosperm) which is available to support the germ has suffered some degradation from enzyme (Briggs and Mac Donald, 1983). During malting, there is an improvement in the nutritional content of grains (Collins and Sanders, 1976). Through the activity of enzymes, some nutrients are degraded (anti nutritional factors inclusive) while others are changed from one form to another.

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OCHEME, O.B et al: Continental J. Food Science and Technology 2: 14 – 19, 2008 The objective of this study is to determine the effect of malting and soybean supplementation on the nutritional value (with respect to proximate composition and amino acid content) and acceptability (with respect to sensory properties) of maize grit porridge (eko-eda). MATERIALS AND METHODS Maize grains (white variety) and soybean seeds were obtained from central market, Minna, Niger State, Nigeria on 7th July, 2007. Preparation of samples Maize grains and soybean seeds were winnowed and hand picked to remove pieces of cobs, stones, damaged seeds and rodents’ droppings. They were then washed with tap water and oven-dried at 600C for 1h. Table 2: Amino acid composition (g/100g) of maize grit and blends of maize-soybeans grits

Grit samples FAO/WHO (1991) Req. Amino acid C MMG MMSG MMMSG Pattern for 10-12year olds Essential Threosine 3.3+0.00 3.3+0.00 3.7+0.01 3.7+0.00 2.8 Cystine 2.0+0.02 2. 0+0.01 1.5+0.00 1.5+0.01 1.2 Valine 4.8+000 4.7+0.00 6.3+0.00 6.3+0.01 2.5 Methionine 1.6+0.00 1.6+0.00 1.2+0.00 1.2+0.01 1.3 Isolencine 4.0+0.01 3.9+0.00 3.8+.0.01 3.8+0.02 2.8 Leucine 11.7+0.00 10.9+0.02 7.4+0.00 7.1+0.01 4.4 Tyrosine 3.5+0.02 3.5+0.01 4.6+0.00 4.7+0.02 1.1 Phenylalanine 5.2+0.01 5.0+0.00 54+0.02 5.4+0.02 1.0 Lysine 3.1+0.00 3.2+0.01 4.7+0.01 4.7+0.02 4.4 Tryptophan 0.5+0.00 0.5+0.01 0.8+0.00 0.8+0.01 0.9 Non Essential Aspartic acid 5.9+0.02 6.1+0.00 7.2+0.02 7.3+0.02 Serine 4.7+0.01 4.7+0.01 4.8+0.00 4.8+0.02 Glutamic acid 15.2+0.02 15.3+0.00 16.6+0.02 16.6+0.01 Proline 7.4+0.01 7.2+0.01 3.3+0.00 3.2+0.00 Glycine 3.7+0.00 3.7+0.01 3.7+0.00 3.7+0.02 Histidine 2.6+0.01 2.8+0.02 3.2+0.01 3.1+0.01 Arginine 5.0+0.00 5.2+0.00 6.9+0.02 6.9+0.01 Alanine 7.5+0.01 7.5+0.00 8.1+0.02 8.2+0.00

Values are mean + SD of triplicate determinations. C = Untreated maize grits MMG = Malted maize grits MMSG = 70:30 blend of untreated maize and malted soybean grits. MMMSG = 70: blend of malted maize and malted soybean grits. Preparation of maize grits A 500g lot of maize grains were dry milled using a double disc attrition mill. The resultant grits were sieved using a 2mm sieve to obtain the desired grit particle size. This was then soaked in water for 2h to

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OCHEME, O.B et al: Continental J. Food Science and Technology 2: 14 – 19, 2008 soften and the germ and hulls were separated from the grits (endosperm) by decanting. The grits were then dried at 1000 C for 1h 30 min in an air oven (Gallenkamp, 3000 plus series, England) and packaged in low density polyethylene. Preparation of malted maize grits A 500g lot of maize grains were cleaned as described previously. The grains were then soaked for 16h with the soak water been changed after 4h. At the end of soaking, the soak water was drained off and the grains were spread evenly on trays and covered with jute bags in a dark, secluded area for 72 hours at 32+ 20C. At the end of malting, the grains were dried at 1000C for 1h 30 min in an air oven. After drying, the grains were devegetated, i.e. the rootlets were removed by rubbing between palms after which they were milled, sieved, soaked in water for 2h to soften. The germ and hull were then separated from the grits by decanting. The grits were then dried and packaged as with the previous sample. Preparation of soybean grits A 500g lot of soybean seeds were malted, dried at 600C for 1h, milled and packaged as was the case with malted maize grits. Soybean was malted primarily to reduce its characteristic beany flavour. Formulation of blends Table 3: Mean sensory scores of eko-eda prepared from maize grits and blends of maize and soybean grits

Samples Parameters C MMG MMSG MMMSG Appearance 6.40a 6.50a 3.70a 4.60ab Colour 6.30a 6.40a 4.20b 5.10ab Taste 5.80a 6.10a 3.80b 4.20ab O/A 6.10a 6.20a 3.90b 4.20ab

Mean values in the same row with different superscripts are significantly different (p<0.05). KEY C = Maize grits (control) MMG = Malted maize grits MMSG = 70:30 blend of untreated maize grits and malted soybean grits MMMSG = 70:30 blend of malted maize grits and malted soybean grits. O/A = Overall acceptability. Values are based on a 7 – point scale where 7 = like very much and 1 = dislike very much. Formulation of blends Malted soybean grits were blended with untreated maize grits in a ratio 30:70 since this was the most organoleptically acceptable level of blending (Ocheme and Alashi, 2007). Malted soybean and malted maize were also blended in the same ratio.

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OCHEME, O.B et al: Continental J. Food Science and Technology 2: 14 – 19, 2008 Determination of Chemical Composition Each sample was analyzed for proximate composition in triplicate. Moisture, protein, fat and ash were determined according to the AOAC (2000) methods. The mineral composition was determined according to the AOAC (1995) method. Amino acid was analyzed according to the method described by Hussain and Basahy (1998). The samples were homogenized in 6ml, 6M HCL for 24h at 1100C. After hydrolysis, the hydrolysate was filtered through a G3 glass filter, evaporated in a vacuum at 400C, then diluted with a citrate buffer (pH 3.2) and analyzed using the amino acid analyzer (SYK 3540). Sensory Evaluation. Eko-eda was prepared from untreated maize grits; malted maize grits; 30:70 blend of malted soybean grits and untreated maize grits; and 30:70 blend of malted soybean grits and malted maize grits. The products were judged by a 20-member panel consisting of staff and students of the Department of Food Science and Nutrition, Federal University of Technology, Minna, Nigeria. A 7-point hedonic rating scale was used where 7 = like very much and 1 = dislike very much. Statistical analysis All data were tested with ANOVA and Duncan multiple range test using the statistical package for social scientist (SPSS, 2000). RESULTS AND DISCUSSION The results of the chemical composition of maize grits and blends of maize-soybean grits are shown in Table 1. Malting and supplementation with soybean significantly (p<0.05) increased the protein and ash content of the grits while significantly (p<0.05) decreasing the fat, crude fibre and carbohydrate content. The increase in protein may be due to the high protein content of soybean which supplemented that of maize while the increase in ash may be due to effect of malting which could have made some hitherto unavailable minerals available due to the degrading action of enzymes. The decrease in fat content may be due to the action of lipolytic enzymes which may have been liberated during malting. It may also be due to utilization of some of the fat to support respiration (Briggs and MacDonald 1983). This reason may also be responsible for the decrease in carbohydrate as a result of malting and supplementation with soybean. Supplementation with soybean significantly (p<0.05) increased the copper, iron and phosphorus content of the grits. Soybean is not a major source of minerals (McGilvery, 1984) but, when included in a mixed diet, contribute to overall requirements. The amino acid composition of maize grits and blends of maize soybean grits is shown in Table 2. Supplementation with soybean caused the lysine content of grits to meet FAO/WHO (1991) requirement pattern. Supplementation also reduced the leucine content of the grits. This is important since excess leucine interferes with the absorption and utilization of isoleucine (Benton et al., 1956). This will result in a better balance of the amino acids. The amino acid profile of maize-soybean grits shows that methionine and tryptophan are the limiting amino acids while other essential amino acids are adequate. The mean sensory scores of eko-eda prepared from maize grits and blends of maize soybean grits are shown in Table 3. The sensory evaluation result showed that there were significant (p<0.05) differences between the eko-eda prepared from maize grits and eko-eda prepared from soybean grits with respect to colour, taste and overall acceptability. This could be attributed to the colour and flavour of soybeans which may have caused a deviation from the conventional eko-eda. However, eko-eda prepared from malted maize grits; blends of untreated maize-soybean grits; and malted maize-soyabean grits were as acceptable as the control sample. CONCLUSION Supplementation with soybean improved the protein, mineral and amino acid content of eko-eda. However, methionine and tryptophan were limiting. Furthermore, eko-eda prepared from blends of maize and soybean grits were acceptable.

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OCHEME, O.B et al: Continental J. Food Science and Technology 2: 14 – 19, 2008 REFERENCES AOAC (1995). Official Methods of Analysis, 15th ed. Arlington, VA: Association of Official Analytical Chemists. AOAC (2000). Official Methods of Analysis, 17th ed.Washington DC Association of Official Analytical Chemists. Benton, D.A; Harper, A.E. and Elvehjem, C.A. (1956) Effect of isoleucine supplementation on the growth of rats fed zein or corn diets. Arch. Biochem. Biophys. (57): 13-19. Bressani, R.; Elias, L.G. and Graham, J.E. (1968). Supplementation con amino acids del maiz de la tortilla. Arch. Latinoam, Nutr. 18: 123-134. Brigg, D.E. and MacDonald, J. (1983). Patterns of Modification in malting. Journal of the Institute of Brewing (89) 260-273 Collins, J.L. and Sanders, G.C. (1976). Changes in trypsin inhibitory activities in some soybean varieties during maturation and germination. J. Food Science (41) 168-172. FAO/WHO (1973). Energy and Protein requirements.FAO Nutrition Meeting Reports Series No. 52; WHO Technical Reports Series No. 522 Rome. FAO/WHO (1991). Protein Quality Evaluation; Food and Agricultural Organisation of the United Nations: Rome, Italy p.66 FAO (1997). Maize in Human Nutrition: Food and Agricultural Organisation, Food and Nutrition Series, Rome/Italy. Hussain, M.A. and Basahy, A.Y. (1998). Nutrient Composition and amino acid pattern of cowpea (Vigna unguiculata(L) walp) grown in Gizan area of Saudi Arabia. Int. J. Food Science and Nutrition, 49, 117-124. McGilvery, R.N. (1984). Nutrition: Minerals and Vitamins in Biochemistry: A functional approach, London: W,B. Sanders Company pp 797-805. Ocheme, O.B. and Alashi, A.M. (2007). Sensory Properties of “eko-eda” prepared from blends of maize and soybean. Proceedings of the 31st Annual conference/General meeting of Nigerian/Institute of Food Science and Technologist. P 114-115.

Received for Publication: 21/02/2008 Accepted for Publication: 13/03/2008 Corresponding Author: OCHEME, O.B Department of Food Science and Nutrition, Federal University of Technology, P.M.B. 65, Minna, Niger State, Nigeria. E-mail: ocheme4ochemeb@yahoo.com

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Continental J. Food Science and Technology 2: 20 – 26, 2008 © Wilolud Online Journals, 2008.

CHEMICAL AND NUTRITIONAL EVALUATION OF THE RAW SEEDS OF MACROTYLOMA CHRYSANTHUM

A. P. UDOH1, E. O. AKPANYUNG2* and I. A EFFIONG1

1Department of Chemistry and 2Department of Biochemistry. University of Uyo. Uyo. Nigeria.

ABSTRACT The paper presents chemical and nutritional information on the underutilised seeds of M. chrysanthum. Chemical analyses show that the seeds are not rich in lipids and hence have a low energy value. The seeds were found to be rich in some essential amino acids (especially lysine), macroelements (Na, K, Mg) and ascorbic acid. The antinutrient levels of tannin, phytic acid and hydrogen cyanide were found to be low hence the seeds could be considered to be nutritionally good. KEYWORDS: Amino acids, Lipids, Macrotyloma chrysanthum, Mineral elements, Proteins, Proximate composition.

INTRODUCTION A reasonable percentage of the population of developing countries suffers from protein malnutrition arising from inadequate intake. In Nigeria today, there is an increase in the monthly earnings of workers but there is also an unprecedented increase in the prices of common food substances such that adequate protein supply is still a serious problem to the citizenry. Although there is an acute need to meet daily nutritional requirements, available rich and cheap sources of nutrients are either unexplored or were abandoned with the advent of modernity and civilization. For sometime now efforts have been directed at identifying alternative and cheap sources of protein (Enobong and Carnovali, 1992, Udoh et al., 1995a, b; Udoh and Akpan, 1997). Macrotyloma chrysanthum (A. Chev.) Verdc. is a leguminous plant whose seeds were widely consumed in Eastern Nigeria up to the late sixties. The cooked seeds of this plant were eaten with cooked or roasted plantain, cocoyam, yam, water yam and the three leaved yam. However, with the increasing cultivation of various varieties of cowpea, this food has been neglected. It’s cultivation is not given priority and is almost extinct in some areas. In the present study, an attempt is made to decipher the chemical composition of the seeds of M. chrysanthum and to understand its nutrient potential. MATERIALS AND METHODS Mature dry pods of M. chrysanthum were collected from a local farm at Ukana Ikot Ideh, Essien Udim Local Government Area, Akwa Ibom State, Nigeria. The seeds were separated from the pods and dried in an oven at the temperature of 500C for 24 hours. The seeds were then pulverized into fine powder using a clean and dry electric blender. The powder obtained was stored in an air tight plastic container and used for analyses. Moisture content was determined by drying 40 transversely cut seeds in an oven at 600C and the value expressed as a percentage. Crude protein content was calculated by multiplying the percentage Kjedldahl nitrogen (Humpries, 1956) by a factor of 6.25. Crude lipid, crude fibre and the ash contents were analysed by the (AOAC, 2000) methods. The nitrogen free extract (NFE) or total crude carbohydrate was obtained

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by difference. The energy content of the seeds was estimated in kJ by multiplying the percentages of crude protein, crude fat and NFE by factors of 16.7, 37.7 and 16.7 respectively and taking the sum of the products. TABLE 1. PROXIMATE, MINERAL AND ANTINUTRIENT COMPOSITION OF THE SEED OF M. chrysanthum

Proximate Values, g/100g Moisture (wet weight) 15.80 ± 0.02 Crude fibre 9.30 ± 0.42 Crude protein 23.19 ± 0.01 Crude fat 2.70 ± 0.04 Ash 5.53 ± 0.02 Carbohydrate 59.28 Calorific value kJ/100g 1479.04 Mineral composition, mg/100g Sodium 6055.57 ± 27.49 Potassium 990.00 ± 0.41 Calcium 76.67 ± 0.59 Magnesium 2571.67 ± 23.45 Zinc 35.55 ± 1.25 Copper 1.18 ± 0.14 Iron 26.28 ± 0.02 Manganese 3.75 ± 0.10 Cobalt 0.07 ± 0.01 Cadmium ND Chromium ND Antinutrients, mg/100g HCN 1.19 ± 0.03 Oxalate 8.36 ± 0.11 Phytic acid 8.70 ± 0.01 Tannins 1.55 ± 0.10 Vitamins, mg/100g Ascorbic acid 94.65 ± 0.50

*Each value represent the mean + SD for triplicate determinations

A known mass of the dry powdered sample was ashed at 6000C in a muffle furnace for 4 hours. The ash was dissolved in 6M HCl, made to a definite volume and used for mineral analysis. Sodium and potassium were determined with a flame photometer (Jenway PF 7 Flame photometer, Essex, UK) while other mineral elements were determined using an atomic absorption spectrophotometer (Unicam Analytical System, Model 919, Cambridge, UK). Total oxalate was determined by the method of Dye (1956). Hydrocyanic acid was estimated by the alkaline titration method (AOAC, 2000). The colorimetric method of (Burns, 1971) was used for the determination of tannic acid while phytic acid was estimated with the method of (Wheeler and Ferrel, 1971). Vitamin C was determined by the indophenol reduction method (AOAC, 2000). Amino acids were determined with an automatic amino acid analyzer using the principles of (Moore, 1963) and (Spackman et al., 1958). The total (true) proteins were extracted by the method of Basha et al. (1976). However, ethanol treatment was omitted in order to save the prolamin fraction. The extracted proteins were

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purified by precipitation with cold 20% trichloroacetic acid (TCA) and estimated by the method of (Lowry et al., 1951). The methods of AOAC (2000) were used in the determination of acid value, saponification value, iodine number, peroxide value, ester value, % free fatty acid and unsaponifiable matter. RESULTS AND DISCUSSION The data on proximate composition of M. chrysanthum seeds are shown in Table 1. The seeds contain higher amounts of moisture than the 10% obtained for cowpea and pigeon pea (Sinha, 1977) and 5.16% obtained for the seeds of X. xylocarpa (Siddhuraju et al., 1995). The crude protein content is comparable to TABLE 2. AMINO ACID COMPOSITION OF M. chrysanthum SEEDS.

Amino acid M. chrysanthum Whole Hen’s egg (Paul et al., 1976)

Amino Acid score on Paul et al., 1976)

Isoleucine 5.2 5.6 92.86 Leucine 3.6 8.3 43.37 Lysine 7.7 6.2 124.19 Methionine 1.0 3.2 31.25 Cystine 0.3 1.8 16.67 Phenylalanine 4.8 5.1 94.12 Tyrosine 0.5 4.0 12.50 Threonine 4.1 5.1 80.39 Tryptophan ND 1.8 - Valine 4.0 7.5 53.30 Arginine 7.6 6.1 1.24 Histidine 3.4 2.4 1.41 Alanine 2.9 5.4 0.53 Aspartic acid 8.6 10.7 0.80 Glutamic acid 17.6 12.0 1.46 Glycine 1.1 3.0 0.36 Praline 2.9 3.8 0.76 Serine 4.4 7.9 0.55

values obtained for the seeds of the more common legumes while the contents of fat, fibre and ash were slightly higher than values reported for the seeds of other commonly cultivated legumes (Sinha, 1977). Mineral analysis (Table 1) revealed that the levels of Zn, Fe and Mn were higher than their contents in some of the commonly cultivated legumes (Sinha, 1977; Siddhuraju et al., 1995). The seeds of M. chrysanthum are, however, rich source of Na, K and Mg. Although food legumes are important sources of dietary protein in developing countries, their acceptability and utilization is limited by the presence of relatively high concentrations of some anti-nutritional factors (Nowacki, 1980). Some of these antinutrients like cyanides and tannins are heat labile (Liener, 1980) whereas toxic amino acids, cyanogenic glucosides, saponins, flavones, isoflavones and alkaloids are heat stable (Nowacki, 1980). The data on the antinutrient levels in M. chrysanthum seeds are included in Table 1. The level of tannin appears to be low compared with other cultivated legumes such as green grain, cowpea, pigeon pea, black grain and X. xylocarpa (Khan et al., 1979; Rao and Deosthale, 1982; Siddhuraju et al., 1995). The amount of tannin in M. chrysanthum seed is, however, higher than that found in lima beans, 0.59 mg/100g (Egbe and Akinyele, 1990) but less than that in horse eye bean, 82 mg/100g (Osaniyi and Eka, 1978). The presence of tannins in foods even at low level, is undesirable from nutritional point of view. Tannins are known to inhibit the activities of digestive enzymes (Jambunathan and Smith, 1981).

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A. P. UDOH et al: Continental J. Food Science and Technology 2: 20 – 26, 2008 Soaking and cooking are reported to reduce the levels of phenols, cyanides and tannins in foods (Sathe and Salunke, 1984). This may not apply in the case of M. chrysanthum because the seeds are not usually soaked before cooking and even after cooking the water is not usually discarded. The phytic acid content of M. chrysanthum is also very low compared with the value of 361 mg/100g in the seeds X. xylocarpa (Siddhuraju et al., 1995); 135.8 mg/100g in the seeds of V. unguiculata and 13.5 mg/100g in bambara groundnut (Chakraborty and Eka, 1978). Phytate is known to lower the availability of essential mineral elements from food (Reddy et al., 1982). The seeds of M. chrysanthum contain a low amount of oxalate when compared to the value of 43.0 mg/100g for soyabean seeds (Eka, 1977) and 42.2 mg/100g for the seeds of cowpea (Aremu, 1989). The present study also reveals the presence of HCN in the seeds of M. TABLE 3. CONTENT OF TOTAL (TRUE) PROTEIN AND PROTEIN FRACTIONS OF M. chrysanthum SEEDS.

Protein fraction g/100g seed flour* g/100g seed protein

Total (true) protein 9.60 ± 0.42 100.00 Albumins 0.40 ± 0.27 4.17 Globulins 6.20 ± 0.05 64.58 Prolamins 0.10 ± 0.02 1.04 Glutelins 2.90 ± 0.08 30.21

* Values represent the mean + SD for triplicate determinations chrysanthum. However, the observed level of HCN is much lower than those found in safe varieties of pigeon pea, 86.19 mg/100g; soyabean, 71.81 mg/100g; locust bean, 26.46 mg/100g; bambara nuts, 88.24 mg/100g (Chakraborty and Eka, 1978) and X. xylocarpa seeds, 1.65 mg/100g (Siddhuraju et al., 1995) and may not pose health problems. The ascorbic acid content of M. chrysanthum seeds (94.65 mg/100g) was found to be higher than that of bambara nuts, 1.0 mg/100g (Hepper, 1970) and raw groundnut seeds – 5.8 mg/100g (Elegbede, 1998). Hence, the seeds of M. chrysanthum could be classified as a rich source of ascorbic acid. The amino acid profile of the total seed proteins and the amino acid score are presented in Table 2. The data reveal that the sulphur containing amino acids, methionine and cysteine are the limiting amino acids. As a source of amino acids the seeds of M. chrysanthum are generally poorer than whole hen’s egg (Paul et al., 1976). However, with respect to the contents of isoleucine and lysine, the seeds are quite comparable to whole hen’s egg. It is also noteworthy that the levels of isoleucine, lysine, phenylalanine and threonine obtained in this study compare favourably with those obtained for the seeds of S. stenocarpa (Nwokolo, 1987). Generally, the seeds of M. chrysanthum can be regarded as a moderate source of amino acids and need to be supplemented with other protein rich foods. Seed protein fractionation of M. chrysanthum (Table 3) shows that the globulins and glutelins together constitute the bulk of seed protein (94.8%) as is the case with many other legumes (Boulter and Derbyshire, 1976; Rajaram and Janardhanan, 1991; Siddhuraju et al., 1995). The percentage distribution of the globulin fraction obtained in this study is higher than the value of 54.8% reported for the seed protein of X. xylocarpa (Siddhuraju et al., 1995). The ratio of albumin to globulin is, however, lower than that of X. xylocarpa (Siddhuraju et al., 1995). Data on the chemical characteristics of the crude lipid of M. chrysanthum seeds are shown in Table 4. The iodine value of oil is low and this is an indication of the low degree of unsaturation of the fatty acids present in the oil (Etuk, 2000). However, the iodine value of M. chrysanthum is comparable to that of palm oil which ranges between 37 to 57. Since the iodine value of the seed oil from M. chrysanthum is low, the oil can be classified as a non-drying oil (Ihekoronye and Ngoddy, 1985).

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A. P. UDOH et al: Continental J. Food Science and Technology 2: 20 – 26, 2008 The saponification value gives an estimate of the mean molecular weight of the fatty acids present in the crude lipid. The value obtained in the present study is lower than those of commonly cultivated oil seeds such as groundnut and cotton seed (195) and guar meal (205) (Singh and Misra, 1981). The acid value of M. chrysanthum seed oil is high compared with the reported values for other edible oils such as groundnut oil – 2.0; guar meal – 3.0; and cotton seed oil – 0.5 (Singh and Misra, 1981). The ester value of an oil gives some insight into the total glycerides present whereas the level of free fatty acids indicate the quality of an oil; the higher the amount of free fatty acid the lower the quality of the oil (Etuk, 2000). The peroxide value of an oil is an index of the degree of peroxidation of the oil. The peroxide value obtained is high. This lipid oxidation and its resultant flavour impairment seriously limits the storage potential of the seed oil of M. chrysanthum. TABLE 4. CHEMICAL CHARACTERISTICS OF THE OIL OF M. chrysanthum SEEDS.

Parameter Content* Acid value 19.35 ± 0.28 Saponification value 109.40 ± 0.00 Iodine value 45.30 ± 0.13 Ester value 90.05 ± 0.28 Peroxide value 83.33 ± 3.30 % Free fatty acid 9.73 ± 0.14 % Unsaponifiable yield 5.80 ± 0.10 Unsaponifiable matter 49.54 ± 0.40

* Values represent the mean + SD for triplicate determinations CONCLUSION In conclusion, the present study has shown that the seeds of M. chrysanthum are rich in critical nutrients. It’s nutrient potential compares favourably with those of the more commonly consumed legumes. The antinutrient levels of tannin, oxalate, hydrogen cyanide and phytic acid in these seeds are reasonably low. There is, therefore, the need to revisit the consumption of these seeds in the absence of the better known food legumes. REFERENCES AOAC (2000). Official Methods of Analysis. 17th edn. Association of Official Analytical Chemists, Washington DC, USA Aremu CY (1989). Quantitative estimation of the dietary contribution of phytate, oxalate and hydrocyanate by six popular Nigerian foodstuff. Nig J Nutr Sci 10:79-84 Basha SMM, Cherry JP. Young CT (1976). Changes in free amino acids, carbohydrates and proteins of maturing seeds from various peanut (Arachis hypogaea L.) cultivars. Cereal Chem 53:586-597 Boulter D, Derbyshire E (1976). The general properties, classification and distribution of plant proteins. In: Plant Proteins. Norton G (ed). Butterworths, London Burns RE (1971). Methods of estimation of tannin in grain sorghum. Agronomy J 163:511-519 Chakraborty RE, Eka OU (1978). Studies on hydrocyanate, oxalate and phytic acid content of food stuffs. West Afr J Biol Appl Chem 21:50-59

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A. P. UDOH et al: Continental J. Food Science and Technology 2: 20 – 26, 2008 Dye WB (1956). Chemical studies on Halogeton glomeratus. Weeds 4:55-60 Egbe IA, Akinyele IO (1990). Effect of cooking on the antinutritional factors in Lima beans (Phaseolus lunatus). Food Chem 35:81-87 Eka OU (1979). Studies on the level of oxalic and phytic acid in traditional foods of Northern Nigeria. West Afr J Biol Appl Chem 20:45-54 Elegbede AJ (1998). Legumes. In: Nutritional Quality of Plant Foods. Osagie A, Eka OA (eds). Post Harvest Research Unit. Department of Biochemistry, University of Benin. Benin. Nigeria Enebong HN, Carnovalli E (1992). A comparison of the proximate, mineral and amino acid composition of some known and lesser known legumes in Nigeria. Food Chem 43:169-175 Etuk EUI (2000). Food Biochemistry. Afahaide and Bros Printing and Publishing Company, Uyo, Nigeria Hepper FN (1970). Bambara Groundnut. Field Crops. Abstract 23:1 Hill JW (1988). Chemistry for the Changing Times. 5th edn. Macmillan Publishing Company, New York Humphries EC (1956). Mineral composition and ash analysis. In: Modern Methods of Plant Analysis. Vol. 1. Peach K, Tracey MV (eds). Springer Verlag, Berlin, Germany Ihekoronye AI, Ngoddy PO (1985). Integrated food science and technology for the tropics. Macmillan Publishers Ltd, Lagos, Nigeria Jambunathan R, Singh U (1981). Grain quality of pigeon pea. In: Proceedings of The International Workshop on Pigeon Peas. Vol. 1. ICRISTAT, Pantencheru, Andra Pradesh, India Khan AM, Jacobson I, Eggum OB (1979). Nutritive value of some improved varieties of legumes. J Sci Food Agric 30:395-400 Liener IE (1980). Heat labile antinutritional factors. In: Advances in Legume Science. Summerfield RJ, Bunting AH (eds). Royal Botanic Gardens, Kew, Richmond, Surrey, UK Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951). Protein measurement with folin – phenol reagent. J Biol Chem 193:265-275 Moore S (1963). The determination of cystine as cysteic acid. J Biol Chem 238:235-237 Nowacki E (1980). Heat stable anti-nutritional factors in leguminous plants. In: Advances in Legume Science. Summerfield RJ, Bunting AH (eds). Royal Botanic Gardens, Kew, Richmond, Surrey, UK Nwokolo E (1987). A nutritional assessment of African yam bean (Sphenostylis stenocarpa) and Bambara groundnut (Voandzea subterranea). J Sc Fd Agric 41:123-129 Osaniyi CB, Eka OU (1978). Studies on chemical composition and nutritive value of horse eye bean. West Afr J Biol Appl Chem 21:60-66 Paul AA, Southgate DAT, Russel J (1976). First supplement to McCance and Widdowson’s The Composition of Foods. Her Majesty’s Stationery Office, London, UK Rajaram N, Janardhanan K (1991). The biochemical composition and nutritional potential of the tribal pulse, Mucuna gigantean (Willd) DC. Plant Foods Hum Nut 41:45-51

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A. P. UDOH et al: Continental J. Food Science and Technology 2: 20 – 26, 2008 Rao PV, Deosthale YG (1982). Tannin content of pulses; varietal differences and effects of germination and cooking. J Sci Food Agric 33:1013-1016 Reddy NR, Sathe SK, Salunkhe DK (1982). Phytate in legumes and cereals. Adv Food Res 28:1-92 Sathe SK, Salunkhe DK (1984). Technology of removal of unwanted components of dry bean. CRC Crit Rev Food Sci Nutr 21:263-268 Siddhuraju P, Vijayakumari K, Janardhanan K (1995). Nutrient and chemical evaluation of raw seeds of Xylia xylocarpa: an underutilized food source. Food Chem 53:299-304 Singh SP, Misra BK (1981). Lipids of Guar seed meal. Agric Food Chem 29:907-911 Sinha SK (1977). Food Legumes: distribution, adaptability and biology of yield. FAO Plant Production and Protection (III), FAO, Rome Spackman DH, Stein WH, Moore S (1958). Chromatography of amino acids on sulphonated polystyrene resins. An Improved System. Anal Chem 30:1185-1190 Udoh AP, Akpanyung EO, Igiran IE (1995a). Nutrients and antinutrients in small snails (Limicolaria aurora). Food Chem 53:239-241 Udoh AP, Effiong RI, Edem DO (1995b). Nutrient composition of dogwhelk (T. Cattifera), a protein source for humans. Trop Sci 35:64-67 Udoh AP, Akpan EJ (1997). Chemical composition of Potadoma freethi and Coclicella acuta. J Fd Sci Technol 34:540-542 Wheeler EL, Ferrel RE (1971). A method for phytic acid determination in wheat and wheat fractions. Cereal Chem 48:312-320

Received for Publication: 08/01/2008 Accepted for Publication: 13/03/2008 Corresponding Author: E. O. AKPANYUNG Department of Biochemistry. University of Uyo. Uyo. Nigeria. Email: ediboy2006@yahoo.com

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Continental J. Food Science and Technology 2: 27 - 32, 2008 © Wilolud Online Journals, 2008.

CHEMICAL COMPOSITION AND FUNCTIONAL PROPERTIES OF KIDNEY

BEAN/WHEAT FLOUR BLENDS.

Okoye J.I1, Nkwocha A.C1 and Agbo A.O2.

1Department of Food Science and Technology, Madona University, Elele Campus, P.M.B 48, Elele, Rivers State, Nigeria. 2Department of Industrial Chemistry, Elele Campus, P.M.B 48, Elele, Rivers State, Nigeria.

ABSTRACT The chemical and functional characteristics of kidney bean and wheat flour blends were examined. The kidney bean flour (KF) was composite with wheat flour (WF) at the levels of 10%, 20%, 30%, 40% and 50%. The flour blends were analysed for their chemical composition and functional characteristics. From the results, the protein content of the blends increased with increasing supplementation with kidney bean flour from 22.74% in 50.50 (KF:WF) to 27.24% in 90:10 (KF:WF) samples, while the carbohydrate decreased. Contrarily, the energy content of the blends increased gradually as the level of fortification with kidney bean flour decreased from 360.60KJ in 90:10 (KF:WF) to 362.15KJ in 50:50 (KF:WF). The results also showed that there were significant differences (p<0.05) in emulsion capacity, and oil and water absorption capacities of the blends. KEYWORDS: Chemical, functional, characteristics, kidney bean-wheat flour mixtures.

INTRODUCTION In most developing countries of the world including Nigeria, where diets are composed mainly of one plant staple food, the occurrence of protein-energy malnutrition especially among children is common. As the cost of producing meat, milk, egg and fish which are foods of high biological value increase, plant proteins offer ready and affordable solution to the problem of a growing protein gap. Low cost, protein-rich and high energy food formulations based on cereal legume mixtures have been suggested (Akobundu and Hoskins, 1987); Marero et al., 1988; Okaka et al., 1992). The enrichment or fortification of traditional cereal based diets with other protein sources such as oilseeds and legumes has received considerable attention. This is because oil seed and legumes proteins are rich in lysine, but deficient in sulphur containing amino acids (Ihekoronye and Ngoddy, 1985). Legumes generally contain relatively high amount of protein than other plant food stuffs. Cereals have a low protein content and are in general deficient in lysine but are adequate in sulphur containing amino acids. Legume proteins are mainly used in food formulations to complement the protein in cereal grains because of their chemical and nutritional characteristics (Enwere, 1998). Kidney bean (Phaseolus vulgaris) a grain legume, is one of the neglected tropical legumes that can be used to fortify cereal-based diets especially in developing countries, because of its high protein content (Akobundu et al., 1992). It is also a rich source of vitamin, minerals and relatively high in crude fibre (NAS, 1979). Kidney bean is one such protein source, which when used in the fortification or enrichment of cereal – based diets could go a long way in improving their nutritional status. The objective of study was to examine the chemical and functional properties of kidney bean-wheat flour blends.

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Okoye J.I et al: Continental J. Food Science and Technology 2: 27 - 32, 2008

Kidney bean seeds

Washing

Soaking in water (for 6h)

Dehulling

Draining and Removal of hulls

Boiling (100oC for 30 min)

Drying (65oC for 6h)

Milling (Attrition mill)

Sieving (Fine sieve, 300µm)

Cooked Kidney bean flour

Packaging

Fig 1: Flow chart for the production of cooked kidney bean flour.

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Okoye J.I et al: Continental J. Food Science and Technology 2: 27 - 32, 2008 MATERIALS AND METHODS Mature kidney bean seeds (Phaseolus vulgaris) and the wheat flour used for this study were bought from local market in Owerri, Nigeria. This research work was carried out in the Department of Food Science and Technology, Madonna University, Elele, River State, Nigeria in November, 2007. Table 1: Flour Blends

Samples KF (%) WF (%) A 90 10 B 80 20 C 70 30 D 60 40 E 50 50

KF = Kidney bean flour; WF = wheat flour Table 2: Means 1, 2 of Proximate Composition of Kidney bean/Wheat flour blends on dry weight basis.

Samples Moisture (%)

Nx5.75 protein (%)

Fat (%) Ash (%) Fibre (%)

Carbohydrate (%)

Energy (KJ/100g)

A 13.32a 27.24a 2.36a 4.12a 4.56a 57.60a 360.60a B 12.12b 27.13b 2.48a 4.08a 4.52a 57.45a 360.64a C 9.03c 26.25c 3.05b 3.98b 3.22b 57.70a 361.64a D 8.86d 23.63d 3.45c 1.66c 2.24c 59.14b 362.13b E 8.68c 22.74c 4.15d 1.33d 1.82d 58.57c 362.15b

1. Values are means of triplicates samples 2. Means in the same column and followed by the same letters are not significantly different

from each other (p>0.05). Preparation of Kidney bean flour The kidney bean flour was prepared according to the method described by Giami and Bekebain (1992) as shown in Figure 1. During preparation, two kilograms of kidney bean seeds which were free from dirts and other foreign materials such as stones, sticks and leaves were weighed, cleaned and soaked in tap water for 6h. After soaking, the seeds were drained, dehulled manually, boiled (100oC, 30min) and dried in the cabinet dryer (65oC, 6h). During drying, the dehulled seeds were stirred at intervals of 30 minutes to ensure uniform drying. The dried seeds were milled (attrition mill) and sieved to pass through a 300 mesh sieve. The cooked kidney bean flour obtained was finally packaged in sealed polyethylene bags due to the hygroscopic nature of the flour until used for blending and analysis. Flour Blending The kidney bean flour (KF) was composite with wheat flour (WF) at the levels of 10%, 20%, 30%, 40% and 50% in a kenwood mixer (Model A 990). There after, the flour blends were individually packaged in sealed polyethylene bags and kept at room temperature until used for analysis. The various flour blends produced are shown in Table 1. Chemical Analysis The moisture, protein, fat, ash and fibre contents of each of the flour blends were determined in tripulates using the methods of AOAC (1990). The carbohydrate was determined by difference (Iherokoronye and Ngoddy, 1985). The food energy was calculated using the Atwater factor 4 x protein, 4 x carbohydrate, 9 x fat (Bryant et al., 1988).

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Okoye J.I et al: Continental J. Food Science and Technology 2: 27 - 32, 2008 Evaluation of Functional Properties Emulsion capacity, water and oil absorption capacities and bulk density of each of the flour blends were determined in tripulates by the methods of Okezie and Bello (1988). Swelling capacity was determined according to the method of AOAC (1990). Statistical Analysis Analysis of variance (ANOVA) was performed on all the data after the analyses to detect significant differences (p<0.05) among the sample values. The turkey test was used in separating significant means. Table 3: Means 1, 2 of functional properties of Kidney bean/wheat flour blends on dry weight basis.

Samples Emulsion capacity (ml/g)

Oil absorption capacity (ml/g))

Water absorption capacity (ml/g)

Bulk density (g/ml)

Swelling capacity (%)

A 9.62a 6.28a 9.48a 0.98a 257.1a B 9.54b 6.26a 9.4aa 0.96a 256.2a C 9.42c 5.86b 9.36b 0.97a 255.6a D 9.30d 5.62b 9.24c 0.94a 256.6a E 9.22c 4.48c 9.16d 0.96a 256.4a

1. Values are means of triplicates samples 2. Means in the same column and followed by the same letters are not significantly different from

each other (p>0.05). RESULTS AND DISCUSSION The proximate composition of the flour blends are shown in Table 2. The moisture contents of the blends ranged from 8.68% to 13.32%. The differences could be attributed to inadequate drying of kidney bean seeds after soaking and boiling. The protein content of all the flour blends differed significantly (p< 0.05) from each other. The differences were observed because the protein content of the blends increased steadily with increasing content of kidney bean flour (KF). However, kidney beans have been reported to be good sources of protein (Duke, 1981). This addition effect was also observed for ash and fibre. In other words, the ash, fibre and protein contents of the blends increased as the level of kidney flour inclusion increased. However, the opposite effect (subtraction effect) was observed for fat and carbohydrate contents of the flour blends. The results of fat and carbohydrate contents of the blends are generally in agreement with those reported by Akpapunam and Darbe (1994). The energy contents of the blends ranged from 360.60KJ/100g to 362.15KJ/100g. The energy content of the flour blends was generally higher than those reported by Giami et al., (2000). The increase in the energy content of the blends resulted from their high protein and carbohydrate contents. The supplementation of wheat flour (WF) with kidney bean flour (KF) produced the desired effect of increasing the protein content of the blends, which will invariably improve the nutritional quality of the products made from these flour blends. The results of the functional properties of the blends are shown in Table 3. From the results, the emulsion capacity of the blends ranged from 9.22ml/g to 9.62ml/g with samples A and B having the highest values. The variations in emulsion capacity of the blends could be attributed to differences in their globular protein contents (Sathe et al., 1982). Flour with good emulsion capacities will be useful in the preparation of comminuted meat products and analogs. The oil absorption capacity of the blends differed significantly (p<0.05) from each other. They were also higher than those reported by Akubor et al., (2000). However, flours with high oil absorption capacities will perform excellently as meat extenders. The water absorption capacity of the flour blends ranged from 9.16ml/g to 9.48ml/g. The differences could be attributed to the milling process subjected to each of the flour during processing which resulted in starch damage (Wolf, 1970). The high water absorption capacities of the blends will make them useful in the formulation of doughnuts and pancakes. The bulk density of the blends was generally higher than those previously

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Okoye J.I et al: Continental J. Food Science and Technology 2: 27 - 32, 2008 reported for flours and was also not significantly different from each other (p>0.05). The increase in the bulk density of the blends could be due to the cooking treatment adopted during the processing of kidney bean flour (Arkroyed and Doughty, 1982). The high bulk density of the blends will be an advantage in the formulation of confectionery products. The swelling capacity of the blends ranged from 255.60% - 257.10 % The result also showed that there was no significant difference in the swelling capacity of the flour blends (p>0.05). The increase in the swelling capacity of the samples could be due to their high protein contents (Chang and Satter, 1981). The high swelling capacities of the blends will make them useful in the preparation of soups, puddings and sauces. However, the kidney bean/wheat blended flours could be used as protein supplement and as functional ingredients in the formulation of a number of food products. CONCLUSION The enhancement of the nutritional value of wheat and other cereal flours with the inclusion of kidney bean flour could help to alleviate the problem of malnutrition prevalent in rural areas of our society. From the findings of this study, it was observed that the fortification of wheat and other cereal flours with kidney bean flour at a level up 50% would help immensely in improving their nutritional value and functional characteristics. Further studies should be performed on the flour blends to determine their respective protein quality and amino acid composition. ACKNOWLEDGEMENT The authors thank Miss C. A. Nwanosike and Mr. C. P. Udejiofor for their support and contribution to this study. Moral support received form the management of Madonna University, Elele Campus, River State, is highly appreciated. REFERENCES Akobundu, E.N.T. and Hoskins, F.H. (1987). Potential of corn-cowpea mixture as infant food Journal of Food and Agriculture 2: 111-114. Akpapunam, M. A. and Darbe, J, W. (1994). Chemical composition and functional properties of Miaze and bambara groundnut flours for cookie production. Plant foods for Human Nutrition 46: 147-155. Akubor, P. I., Isolokwu, P. C., Ugbane, O. and Animawo, I. A. (2000). Proximate composition and functional properties of African bread fruit kernel and wheat flour blends. Food Res. Inter. 33: 707-712. AOAC (1990). Official Methods of Analysis. Association of Official Analytical Chemists. 15th edn. Washington, D.C. Pp. 205-228. Arkroyed, W. R. and Doughty, J. (1982). Legumes in Human Nutrition. FADI, Rome. Pp. 232-244. Bryan, L. A., Montecalro, J. J. R., Morey, K. S. and Lay, B. (1988). Processing, functional and nutritional properties of Okro seed products. Journal of Food Science. 53: 1399-1402. Chang, K. and Satter, L. (1981). Isolation and characterization of the Major proteins from the Northern Beans. (Phaseolus vulgaris) Journal of food Science 46:462-468. Duke, J.A. (1981): Handbook of Legumes of World Economic Importance. Plenum Publishing Corporation, New York. Pp. 184-220. Enwere, N. J. (1998): Foods of Plant Origin. Afro-Obis Publications Ltd., Nsukka. Pp.43-46.

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Okoye J.I et al: Continental J. Food Science and Technology 2: 27 - 32, 2008 Giami, S. Y. and Bekebain, D. A. (1992). Proximate compositional and functional properties of raw and processed full-fat fluted pumpkin seed (Jelfairia occidentalis) flour. Journal of the Science of Food and Agriculture 59: 321-325. Giami, S. Y., Adindu, M. N., Akusu, M. O. and Emelike, J. N. T. (2000). Composition, functional and storage properties of flour from raw and heat processed African bread fruit (Trecuia Africana Decne). Plant Foods for Human Nutrition 55: 357-368. Ihekoronye, A. I. and Ngoddy, P.O. (1985). Intergrated Food Science and Technology for the Tropics. Macmillan Publishers Ltd. London. Pp. 322-346. NAS (1979): Tropical Legumes: Resources for the Future. National Academy of Science (NAS). Washington. D.C. Pp. 150-168. Okaka, J.C., Akobundu, E. N. T. and Okaka, A. N. C. (1992). Human Nutrition – An Intergrated Approach. Obio Press Ltd., Enugu. Pp. 182-220. Okezie, B. O. and Bello, A. B. (1988). Physiochemical and functional properties of winged bean flour and isolated compared with soy isolate. Journal of Food Science 53: 1450-1454. Sathe, S. K., Deshpande, S. S., and Salunkhe, D. K. (1982). Functional properties of winged bean protein. Journal of Food Science 47: 503-506. Wolf, W. J. (1970. Functional, chemical and physical properties of soybean protein. Journal of Food and Agriculture 14: 969-976. Received for Publication: 08/03/2008 Accepted for Publication: 13/06/2008 Corresponding Author: Okoye J.I Department of Food Science and Technology, Madona University, Elele Campus, P.M.B 48, Elele, Rivers State, Nigeria.

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Continental J. Food Science and Technology 2: 33 - 36, 2008 © Wilolud Online Journals, 2008. CHEMICAL COMPOSITION AND FUNCTIONAL PROPERTIES OF IRVINGIA GABONENSIS SEED

FLOUR

Abulude F.O1, Alo F. I2, Ashafa3 S.L and Fesobi M4. 1Department of General Studies, Federal College of Agriculture, Akure, Ondo State. 2Department of

Metallurgy, Obafemi Awolowo University, Ile Ife, Osun State. 3Department of Animal Production, Federal College of Agriculture, Akure, Ondo State. 4Department of Biochemistry, Adekunle Ajasin University,

Akungba - Akoko, Ondo State

ABSTRACT The Irvingia gabonensis seed has been evaluated with respect to proximate, mineral composition and functional properties. The seed contained (g/100g): protein (12.78), fibre (5.87), fat (40.26) and carbohydrate (37.47). The predominant metal in the seed is sodium (840mg/100g). Water absorption, oil absorption and oil emulsion capacities are relatively high, while foaming capacity and least gelation concentration are low. The results showed that the seed may be useful in some food formulations. KEYWORDS: Irvinga gabonensis, mineral, proximate composition, functional properties.

INTRODUCTION In many rural tropical communities, local-seeds from wild or semi-dome, herb trees and climbers, are used in the diet, often the oil is extracted and the crushed seed is added to soups and stews. An example of one of such seeds is that of the Irvingia gabonensis, sometimes known as”Wild mango” (FAO, 1989). Wild mango is a large tree up to 120ft. high or more, with grey trunk slight buttressed. This tree is found in an evergreen forest and mixed deciduous forest, sometimes seen in villages or towns. The pulp, kernel and fruit are said to be eaten by man and animal, although bitter and acrid, with the flavour of turpentine. The fruits are rich in oil and are important because of their use in making bread, chocolate, cheese butter, soap and feed cake. The wood is suitable for planking of ships decks, wood paving, house post, canoes, pestles, for household utensils and the bark is medicinal (Irvine, 1961). Some authors have reported the proximate compositions, metal contents and production of wine from this seed (Uzegbu, 1993; Ojukwu et al, 2000) elsewhere. However, there is little or no information on the functional properties of the Irvingia gabonensis seed. Therefore, this present study investigated the chemical composition and the functional properties of the seed in an attempt to assess its value for application in the food industries. MATERIALS AND METHODS The sample was obtained from a farm site in Arimogija, Ose Local Government area of Ondo State, Nigeria. The seed was carefully washed with distilled water, sun dried for 3 days, ground with a Kenwood blender and kept in a dry container prior to analyses. Proximate analysis of the sample for moisture, dry matter and crude fat were determined in triplicate using the methods described by AOAC (1990). Nitrogen was determined by the micro-Kjeldahl method described by Pearson (1976), and the result was used for estimating the crude protein content by multiplying by 6.25. The ash content was determined using the method described by Pearson (1976). Carbohydrate was determined by difference (100 – ash + protein + fibre + fat). Minerals were determined using a solution that was obtained by dry-ashing the sample at 550oC and dissolving it in distilled water that contains a few drops of concentrated hydrochloric acid in a volumetric flask. Na and K contents were measured with a Corning 405 flame photometer (AOAC, 1990). Fe, Mg and Ca were determined with a Perkin Elmer 306 atomic absorption spectrophotometer. The

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Abulude F.O et al: Continental J. Food Science and Technology 2: 33 - 36, 2008 Table 1: Proximate and mineral constituents of samples analyzed (n=3)

Parameter Mean±SD

Moisture (g/100g) 8.25 (0.76) Ash ‘ 3.62 (0.22) Protein ‘ 12.78 (0.79) Fibre ‘ 5.87 (0.22) Fat ‘ 40.26 (0.88) Carbohydratea ‘ 37.47 (0.72) Calcium (mg/100g) 560 (3.42) Potassium ‘ 110 (1.22) Sodium ‘ 840 (5.22) Magnesium ‘ 270 (1.98) Iron ‘ 10 (0.15) Phosphorus ‘ 240 (1.97)

n – number of determinations, a – calculated by difference protein solubility of the sample was determined at various pH values by the method described by Oshodi and Ekperigin (1989). The least gelation concentration, water and oil absorption, and foaming properties of the sample were determined by the methods described by Sathe et al (1982). RESULTS AND DISCUSSION Table 1 contains the proximate composition of the seed. The moisture content is low (8.25g/100g). This low value could be an added advantage to the shelf life of the seed. The ash content is low. This was depicted by standard deviation of 0.22. Fibre content (5.87g/100g) is low. Its ingestion will help to reduce blood cholesterol levels and the risk of bowel cancer and gall stones (Taylor et al, 1997). The fat content is high (40.26g/100g). This suggests that sample is a good source of vegetable oil. The carbohydrate content is 37.47g/100g. Carbohydrate makes up more than 82% of starch content. This means that the seed could be a good source of starch of human consumption and/or the industry. The above observation is in line with what was contained in other literature reports on soup thickeners (Uzuegbu, 1993), baobab seeds (Odetokun, 1996) and cooked seeds of locust beans (Adeyeye et al. 2002) Table 1 also depicted the mineral contents of the sample in mg/100g. Sodium with a value of 840mg/100g was found to be the most abundant mineral in the seed. This observation is close agreement with what was reported by Uzuegbu (1993) and Odetokun, (1996). The results calcium, magnesium, phosphorus and potassium contents in our present study are different from the values reported for African yam bean (AYB) (Oshodi et al., (1997); Adeparusi, (2001); Abdullahi; (2002). Iron concentration is the lowest mineral (10mg/100g) in the samples, hence seed may not be a good blood enhancing substance for the treatment of anemic patient. Table 2 shows the results for the functional properties of the sample analysed. The water absorption capacity (WAC) of the seed is 150%. This value compared favourably with WAC reported for some seeds of AYB (Adeyeye and Aye, 1996; Oshodi et al., 1997). Cola acuminata (Abulude, 2002) and breadnut (Nwabueze et al., 2001). WAC has been very useful in the production of viscous foods. Hence, Irvingia gabonensis seed may be useful in the production of soups, gravies and doughs. Oil absorption capacity (OAC) is found to be 217%. This value is high than the values and for Adenopus breviflorus benth flour (Oshodi, 1992) and AYB (Oshodi et al., 1997). The result is however, similar to those reported by Fagbemi and Olaofe (2000) for precooked cocoyam flour. In food formulation OAC plays a vital role in increasing the mouth feel of foods and acts as flavour retainer (Kinsella, 1976a). Least gelation concentration (LGC) is 10%. This result compared well with values reported in the literature. The LGC

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Abulude F.O et al: Continental J. Food Science and Technology 2: 33 - 36, 2008 low value might have been caused by the high value of the carbohydrate content in the seed. Subsequently, the seed may be useful in the production of curd or as an additive to other materials for forming gel in food Table 2: Functional properties of sample analyzed (%)

Parameter Mean±SD Water Absorption Capacity 150 (1.40) Oil Absorption Capacity 217 (1.98) Least Gelation Concentration 10 (0.42) Foaming Capacity 35 (0.72) Oil Emulsion Capacity 97 (0.53)

Table 3: Protein Solubility (%) as a function of pH pH Protein Solubility 2 82 3 76 4 71 5 45 6 48 7 52 8 65 9 50 10 70 11 76 12 81

product. The values recorded for foaming capacity (FC) in this study is higher than those reported in literature. The high FC will enhance the seeds functionality in the production of cakes and whipping toppings (Kinsella, 1976b). The oil emulsion capacity (OEC) is 97%. This value is higher than that reported for locust beans (Adeyeye et al., 2002). Hence, this seed may be ideal as an additive for the stabilization of emulsion in sausages (Altschul and Wilcke, 1985). Table 3 shows the results of the effect of pH on the solubility of the Irvinga gabonensis seed. It shows that a minimum solubility occurs in pH 5 and 9, and this corresponds to the iso-electric point of proteins and amino acids are least soluble at their iso electric points. CONCLUSION The present results show that the seed has high carbohydrate, fat and mineral contents. The functional properties were good and so this seed may have great potentials for food formulations. ACKNOWLEDGEMENTS The authors are grateful to Mr. Lanre Aregbesola for the provision of the sample used, and to Engr. Aladewolu for production of the manuscript. REFERENCES Abdullahi, S.A. (2002). Evaluation of the nutrient composition of some fresh fish families in Northern, Nigeria. J.Agric. and Environ. 1(2): 141 -150. Abulude, F.O. (2000): Composition and certain food properties of Cola nitida and Cola acuminata flour found in Nigeria. Global J. Pure and Appl. Sci (in press). Adeparusi, E. O. (2001): Effect of processing on some minerals, anti-nutrients and nutritional composition of African yam bean. Sustain Agric. Environ. 3(1): 101 – 108.

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Abulude F.O et al: Continental J. Food Science and Technology 2: 33 - 36, 2008 Adeyeye, E.I. and Aye, P.A. (1998): The effects of sample preparation on the proximate composition and the functional properties of African Yam Bean (Sphenostylis sternocarpa Hochst ex A. rich) flours. La Riv. Ital. Delle. Sost. Grasse. 55:253- 262. Adeyeye, E.I., Ipinmoroti, K.O. and Oguntokun, M.O. (2002): Chemical composition and functional properties of African locust bean (Parkia biglobosa) seeds. Pak J. Sci. Ind. Res. 45 (1): 29-33. Altschul, H. M. and Wilcke, H. L. (1985): New protein foods, Vol. 5, seed storage Protein, Academic Press, London. AOAC (1990): Official Methods of Analysis, Washington, D. C. Association of Official Analytical Chemists. Fagbemi, T.N. and Olaofe, O. (2000): The chemical composition and functional properties of raw and precooked taro (Colocasia esculenta) flours. J. Bio. & Phy. Sci. 1:98-103. FAO (1989): Utilization of tropical foods: tropical oil seeds. Nutrition paper 47/5. Food and Agricultural organization of the United National, Rome. Irvine, F.R. (1961): Woody plants of Ghana. Oxford University Press, London. Pp 506- 507. Kinsella, J. (1976a): Functional properties of protein foods. Crit. Rev. Fd. Sci. Nutri. I: 210. Kinsella. J. (1976b): Functional properties of soy proteins J. Am. Oil Chem. Soe.6:212- 258. Nwabueze, T. U; Ugaduh, A and Nwabueze, J. C. (2001): Functional properties of breadnut flour. J. Sustain. Agric. Environ. 3(2): 93-100. Odetokun, S.M. (1996): The nutritive value of Boabab fruit (Andasonia digitata). La Riv. Ital. Delle Sost. Grasse 53: 371-374. Ojukwu, U.P; Enujiugha, I.B. and Nzejiofor. E. N. (2000): production of wine from wild mango (Irvingia gabonensis) pulp. Nig. J. Tech. Educ. 17(1&2): 51-56. Oshodi, A.A. and Ekperigin, M.M. (I989): Functional properties of pigeon pea (Cajanus cajan) flour. Food Chem. 36: 187-191. Oshodi, A.A. Ipinmoroti, K.O. and Adeyeye, E.I. (1997): Functional properties of some varieties of African yam bean (Sphenostylis stenocarpa) flour – III. Int. J. Food Sci. & Nutr. 48: 243-250. Pearson, D. (1976): Chemical analysis of foods: 7th Edn. Churchill Living stone, London. Sath, S.K; Deshpamde, D.E.S. and Salankhe, D.H. (1982): Functional properties of the great Northern bean (Phascolus vulgaris L.) Protein emulsion, foaming, viscosity and gelation properties. J. Food Sci. 46:71-74. Taylor, D.J., Green, N.P.O. and Stout, G.W. (1997): Biological science 3rd edition Cambridge University Press, UK. Pp 251-252. Uzuegbu, J.O. (1993): Proximate and metal composition of some soup thickeners used in Nigeria. Nig. J. Tech. Educ. 10 (1&2): 57-59. Received for Publication: 08/07/2008 Accepted for Publication: 13/09/2008

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Continental J. Food Science and Technology 2: 37 - 43, 2008 © Wilolud Online Journals, 2008.

CHEMICAL AND MICROBIOLOGICAL EVALUATION OF SOYBEAN FLOURS BOUGHT FROM

LOCAL MARKETS IN ONITSHA, ANAMBRA STATE, NIGERIA.

Eze, V. C1., Okoye J. 12., Agwung, F.D1 and Nnabueke, C1. 1Department of Microbiology, Madonna University, Elele Campus, Rivers State, Nigeria.

2Department of Food Science & Technology, Madonna University, Elele Campus, Rivers State, Nigeria.

ABSTRACT The Chemical and microbiological quality of soybean flours bought from different Local markets in Onitsha, Anambra State were examined. The flour samples were evaluated for their proximate and microbiological quality using standard methods. From the results, the proximate composition of the flours showed significant difference (P < 0.05) in their moisture, protein, fat, ash, carbohydrate and energy contents. The mean microbial counts in the samples from main market were total aerobic bacterial count 6.2 ± 1.4 x 103 and coliform count 2.3 ± 0.8 x 103 cfu/g, E. coli count 1.9 ± 0.7 x 103 and fungal count 7.3 ± 2.9 x 103 cfu/g. The samples from Ose market had the microbial counts as follows: total acerbic bacterial count is 7.8 ± 1.3 x 103 cfu/g; coliform count 3.7 ± 1.2 x 103 cfu/g; E.coli count 2.5 ± 2.7 103 and fungal count 7.0 ± 2.9 x 103 cfu/g. The microbial counts in the samples from Ochanja market were total aerobic bacterial count 4.1 ± 1.9 103 cfu/g; coliform count 2.1 ± 0.4 x 103 cfu/g; E. coli count 1.2 ± 0.4 103 cfu/g and fungal count 3.6 ± 2.4 x 103 cfu/g. The bacteria isolated were Staphylococcus aureus, Streptococcus spp, Escherichia coli, Bacillius spp, Lactobacillus spp, Pseudomonas spp and Micrococcus spp. The fungal genera isolated were Aspergillus, Rhizopus, Penicillium and Fusarium KEYWORDS: Chemical, Microbiological, Soybean Flour, Local Markets.

INTRODUCTION Soybean, which is botanically known as Glycine max (L) Merr, is an important oil seed belonging to the family leguminosae. Soybean is mainly cultivated for its seeds, used as human food, livestock feed and for the extraction of oil. The seeds of various varieties of soybeans may be spherical, elongated and flat in nature, but the industrial varieties are particularly yellowish in colour and oval in shape (Iwe and Onuh, 1992). The production and utilization of soybean as food by humans is associated with the history of China where it had been almost the sole source of protein for generations. The crop has become an increasingly important agricultural commodity in the past several decades, having a steady increase in its annual production, especially in the United States and the world in general. There has been equally a conscientious effort to introduce the crop into poorer regions of the world with reputable marginal malnutrition (Parman, 1974; Danshiell, 1993). Soybean is capable of producing the greatest amount of protein used as food for man. However, this unique plant is generally regarded as an excellent food crop for the protein deficient countries of the world (Liu, 2000). In Nigeria, commendable progress has been made especially at International Institute of Tropical Agriculture (11TA) Ibadan, in the development of different varieties of soybean that could readily adapt to our ecological condition (Solabi, 1993). In spite of these achievements, the potentials of soybean, which is not only rich in oil but contains protein of high biological value have not been adequately exploited in the manufacture of different types of food products in Nigeria. Soybean can be processed into soymilk, soy sauce, tofu (soybean curd), soy-yogurt, soy flour, soy sprouts and many other soy products. Defatted soybean flour can be used for the production of protein isolates and concentrates. Nutritionally, Soybean protein resembles animal protein more closely than other vegetable proteins. Soybean protein constitutes about 40% of the total solids and plays a very important role in food processing

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 (Iwe, 2003). The protein content of soybean and other legumes have a well-recognized deficiency of essential sulphur containing amino acids notably methionine and cysteine but are rich in lysine (Enwere, 1998). It is also a rich source of vitamin, minerals and is relatively low in crude fibre (Ang, 1985). The objective of this study was to examine the chemical and microbiological qualities of soybean flours bought from different local markets in Onitsha, Anambra State. MATERIALS AND METHODS Sources and preparation of samples The samples of soybean flour used for this study were bought from Ose, Ochanga and Main markets in Onitsha, Anambra State. Four samples of the flour were randomly bought from each of the three markets. After purchase, the flour samples were individually packaged in sealed polyethylene bags and kept at room temperature until used for analysis. This research work was carried out in the Department of Microbiology, Madonna University Elele Campus, Rivers State, Nigeria in August, 2007. Chemical reagents The chemical reagents employed in the study were of analytical grade and were products of BDH chemical, Pooles, England and Sigma Chemical Company, St Louis, Missouri, USA. The microbiological media used were products of Oxoid and Difco Laboratories, England. They included nutrient agar used for the estimation of total heterotrophic aerobic bacteria Sabouraud dextrose agar (SDA) used for the isolation of fungi and the MacConkey agar was used for the isolation of coliforms. Chemical analysis The moisture crude protein, fat and ash contents of each sample of the flour were determined in triplicates according to the methods of AOAC (1990). The carbohydrate was determined by different (Ihekoronye and Ngoddy, 1985). The food energy was calculated using the Atwater factor 4 x protein, 9 x fat 4 x carbohydrate (Marero et al., 1988). Enumeration of Coliforms, total heterotrophic bacteria and fungi Samples of the soybean flour were serially diluted in ten folds. Total viable heterotrophic aerobic counts were determined using pour plate technique. Then molten nutrient agar Sabourand dextrose agar and MacConkey agar at 450C were poured into the Petridishes containing 1mL of the appropriate dilution for the isolation of the total heterotrophic bacteria, fungi and coliforms respectively. They were swirled to mix and colony counts were taken after incubating the plates at room temperature for 48h. Characterization and identification of the isolates Bacterial isolates were characterized and identified after studying their Gram reaction as well as cell micromorphology. Other tests performed included spore formation, motility, Oxidase and catalase production, citrate utilization, Oxidative/fermentative ()/F) utilization of glucose, indole product, methyliod-voges Roa-skaur reaction, uease and coaguase production, starch hydrolysis and sugar fermentation. The tests were performed according to the methods described by Gerhardt et al. (1981); Stewart and Beswick (1977); Cruick shank et al. (1980) was performed using the keys provided in the Bergey’s Manual of Determination Bacteriology (1994). Fungal isolates were examined macroscopically and microscopically using the needle mount technique. Then identification was performed according to the scheme of Barnett and Hunter (1972) and Larone (1986). Statistical analysis Analysis of variance (ANOVA) was used where applicable for the detection of significant differences (P< 0.05) among the samples values. The turkey test was used in separating significant means. RESULTS AND DISCUSSION The proximate compositions of the flour samples are shown in Table 1. The moisture content ranged from 9.46% to 10.26%. The differences could be attributed to poor processing methods employed during processing. The protein of all the flour samples differed significantly (P < 0.05) from each other. The

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 variations in the protein content of the flours could be due to differences in heat treatment adopted during processing which affected their level of protein as a result of scorching effect of heat. The protein content of soybean flours is in agreement with those reported by Iwe (2003). The fat content of the flours ranged from 22.44% to 23.86%. The higher fat content of all the samples could be attributed to the fact that the flours were not defeated. However, soybeans have been reported to be good sources of fat (Salunkhe et al., 1992). The ash content of the flours was not significantly different from each other (P> 0.05). They were also similar to those reported by Enwere (1998). The carbohydrate content of the flour samples was significantly different (P < 0.05). The carbohydrate of the flour samples is in agreement with those reported by Fukushima (1991). The energy content of the flours ranged from 455.24 Kcal /100g to 459.62 Kcal/100g.There was significant difference in the energy content of the soybean flours. The energy content of the flour samples was higher than those reported by Okaka et al. (1992). Table 1 means 1,2 of proximate composition of soybean flours on dry weight basis Samples Moisture

(%) N x 5.71 Protein (%)

Fat (%)

Ash (%)

Carbohydrate (%)

Energy (Kcal/100g)

Osm1 9.58a 40.28a 22.60a 4.86a 22.68a 455.24a Osm2 9.54a 40.25a 22.76a 4.82a 22.63a 456.36a (456.36b) Osm3 9.44a 40.16a 23.68b 4.76a 21.96a 461.60C Osm4 10.02b 40.22a 23.60b 4.72a 21.44b 459.04d Om1 9.57a 40.20a 23.86a 4.68a 21.69b 462.30c Om2 9.54a 39.98a 22.74a 4.65a 23.09c 456.94a Om3 10.15b 38.28b 22.52b 4.68a 24.37d 453.28e Om4 9.48a 40.12a 22.44a 4.67a 23.29c 455.60a Mm1 10.26b 40.08a 23.86b 4.66a 21.14b 459.62d Mm2 10.20b 38.68b 22.76a 4.64a 23.72c 459.02d Mm3 10.18b 38.82b 23.52b 4.68a 21.74b 457.92f (459.92f) Mm4 9.46a 40.24a 22.46a 4.74a 23.10c 455.50a Legend: Osm= Ose market; Om= Ochanja market; Mm= Main market 1.Values are means of triplicate determination 2.Means in the same column and followed by the same letters are not significantly different from each other (P > 0.05) The increase in energy content of flours could be attributed to their high protein and fat contents. Table 2 shows the microorganisms isolated and their percentage occurrence. It has been observed that the primary causative agents of microbial spoilage of food are the bacteria, yeast and mould (FAO, 1979). The isolation of these microorganisms from the soybean flour samples is an indication of their involvement in the contamination and spoilage of the product. This also shows evidence of proteolytic and lipolytic activities (Osuji, 1977; Turner, 1983; Agwung et al., 2006). They also reported that the following genera of microorganisms, which include Achromobacter, Flavobacterium, Bacillus, Micrococcus Pseudomonas, Staphylococcus Aspergillus, Rhizopus, Mucor, Fusarium, Neurospora and Candida, are involved in the breakdown of fats oil and protein. The presence of Staphylococcus aureus, which is a normal flora of the body, indicates contamination from handlers. The organism can pass onto the food during harvesting, processing or even storage. The consumer is at risk of acquiring food borne diseases. Staphylococcus is major cause of food poisoning known as Staphylococcal food poisoning. The poisoning is caused by the ingestion of an enterotoxin produced which is characterized by diarrhea and vomiting (Singleton, 1995; Frazier and westhoff, 2004).

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 Bacillus spp, which are Gram-positive, aerobic spore formers, were also present. Most members of the genus are saprophytic organisms prevalent in soil, water and air and on vegetation. Bacillus cereus and Bacillus Subtilis are the most encountered in the group. Bacillus cereus when grown on food causes food poisoning by the production of an enterotoxin (Thomas, 1994,; Brooks et al., 2005). Escherichia coli is an enteric organism and its presence is an indication of faecal contamination of the Samples. This may be attributed to improper sanitary condition during processing of the flour from the water supply, unsterilized utensils and contamination by flies. It causes gastroenteritis especially in infants and young children (Turner, 1983; Brooks et al., 2004). Table 3 shows the mean microbial count of the soybean flour samples. The high mean microbial counts recorded shows exposure of the samples to different genera of bacteria and fungi leading to their contamination. This may also be attributed to high protein and lipid contents as observed in the proximate analysis of the samples. The high counts may also be as a result of the large number of people that visit the markets resulting in increased microbial numbers. The air borne flora may dominate s a result of physical disturbances. The main sources of contamination include humans, sewage, utensils, processing, equipment, handling and storage condition and rodents (Benwart, 2002; Benchat, 1996; Frazier and Westhoff, 2004; Akobundu, 1980). The fungal genera isolated could be traced to harvesting period. Their spouse may have been attached to the gains which overcome the adversely conditions during preparation and finally appeared in the finished products. Table 2: Microorganisms isolated and their percentage occurrence

Organism % occurrence Bacteria Bacillus 13.33 Escherichia coli 20 Staphylococcus aureus 33.33 Streptococci SPP 6.67 Lactobacillus spp 6.67 Protus spp 6.67 Micrococcus 6.67 Psendomonas 6.67 Fungi Rhizopus SPP 50 Aspergillus 25 Penicillium 12.5 Fusariium 12.5

Table 3: The mean microbial counts from soybean flour samples Sample code Total aerobic Coliform count E. coli count Fungal count

Bacterial count (Cfu/g) (Cfu/g) (Cfu/g) (Cfu/g)

MM 6.2 ± 1.4 x 103 2.3 ± 0.8x103 1.9 ± 0.7 x 103 7.3 ± 2.9x 103 OSM 7.8 ± 1.3 x103 3.7 ± 1.2 103 2.5 ± 2 x 103 7.0 ± 2.9 x 103 OM 4.1±1.9 x 103 2.1± 0.4 x 103 1.2 ± 0.4 x 103 3.6 ± 2.4 x 103

Legend: MM = Main Market; OSM = Ose Market; Ochanja Market

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 CONCLUSION Soybean is an important and economically valued food. It is highly nutritious and can be used to solve various malnutrition problems. From the results, it was observed that the source of the samples has no effect on the chemical composition. The bacteria and fungi isolated from the samples could be an indication of health hazards. Therefore microbiological safety should be employed in order to reduce the rate of contamination, which will greatly enhance the quality of the product. The producers, distributors and hawkers of the products should be properly educated on good sanitary practices during the manufacturing, distribution and sale of the product. ACKNOWLEDGEMENT We sincerely thank the Department of Microbiology, Madonna University, Elele Campus for providing all the necessary assistance for the completion of this work. REFERENCES Agwung, F.D., Ifeanyi, V. O., Kanu, Okeke, E. V. and Onome E. D.(2006). Microbial Qualities of Ready to eat meats sold in Elele, Rivers State, Nigeria, International Journal of Biotechnology and Allied Sciences; 1 (1): 14-18 Akobundu, I. O. (1978). Chemical used control in cowpea and soybean in Southern Nigeria, paper prepared for the 3rd symposium on weed control in the tropics, Dakar Senegal volume 2. Ang, G.H. (1985). Nutritional valves of soybean, cowpea and spray dried milk powder. Acro Publishing Inc, New York; pp 233-254. AOAC (1990). Official methods of Analysis. Association of Official Analytical Chemists 15th Edition. Washington, DC.pp.210-225. Barnett, H. L. and Hunter, B. B. (1972). Illustrated genera of imperfect fungi 3rd edition, Burgess publishing company Minniesota,U. S. A. Benchat, I. R. (1996). Pathogenic microorganisms associated with fresh produce, Journal of Food Protection; 59:827 Benwart, G. J. (2002). Basic Food Microbiology, 2nd edition, Van Nostrand Reinbold, New York, pp,665. Bergey’s Manual of Determinative Bacteriology, (1994). 9th edition Holt J. G. (Ed.), Williams and Wilkins Co. Baltimore, pp 783 Brooks, G. F., Butel, S. J. and Morse, S. A. (2004). Medical Microbiology 23rd edition, the McGraw Hill companies Inc. Singapore. Cruickshank, R., Duguid, J. P. and Swain, R. H. A. (1980). Medical Microbiology, 12th edition, the English Language Book Society, E&S Livingstone Ltd, Edinburgh. Danshiell, K.E. (1993). Soybean production and utilization in Nigeria. Paper presented at the national (workshop) on small scale and industrial level processing of soybean organized by IDRC /11TA soybean utilization project, Ibadan. pp. 10-21. Enwere, N.J. (1998) Foods of plant origin. Afro-Orbis Publications Ltd, Nsukka. p 125-240. F. A. O. (1979). Bulletin Publisher, U.N.O.

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 Frazier, W. C. and Westhoff, D. C. (2004). Food Microbiology 4th edition, McGraw Hill Book Co. New York. Fukushima, D. (1991). Recent progress of soybean protein foods. Chemistry, Technology and Nutritional Food Review Int, 7(B): 323-351. Gerhardt, P., Murray, R. G. E., Costilow, R. N., Nester, E. W., Wood, W. A., Kriey, N. R. and Philips, G. G. (1981). Manual of Methods for General Bacteriology, American Society for Microbiology Washington D. C. Ihekoronye, A.1. and Ngoddy, P.O. (1985).Integrated for science and Technology for the Tropics. Macmillan publisher Ltd, London and Oxford. pp 283-292. Iwe, M.O. and Onuh, J.O. (1992). Functional properties of soybean and sweet potato flour mixtures. Lebensm Wiss. U-Technology; 25: 569 - 573.

Iwe, M.O. (2003). Science and Technology of Soybean, Rejoint communication services Ltd, Enugu. pp. 126-162 Larone, D. H. (1986). Important fungi: A Guide to identification, Harper and Row Publishers, Hagerstown, Maryland pp 7-26. Liu, K. (2000). Expanding soybean food utilization,Journal Food Technology 54(7): 46-47. Marero, L. M., Payuma, E. M., Librando, E.C., Lainez, W.N., Gopz, M.D. and Homma, S. (1988). Technology for weaning food formulations prepared from germinated cereals and legumes. Journal of Food Science; 53(5): 1391-1395. Okaka, J. c., Akobundu, E.N.T and Okak, A.N.C. (1992). Human Nutrition – An Integrated Approach. Esut Publication, Obio Press Ltd., Enugu, pp 182-210 Osuji, F. N. C. (1977). The influence of traditional handling on the quality of processed fish in Nigeria. In: Proceeding of conference on the handling processing and marketing of tropical fish, Tropical Product Institute, Lagos, pp 1-75. Parman, G. K. (1974). Agency for international development program for development and utilization of soybean in developing world. J. Am. Oils Chemists’ Society. 51:150-152. Prescott, M. L., John, P.H. and Klein, D. A. (2004). Microbiology 5th edition, McGraw Hill Publishers, New York; 939-941. Salunkhe, D.K, Charan, J.C., Adesule, R. N. and Kadam, S.S. (1992). World’ oil seed, Chemistry, Technology and utilization. An Avi Book published by Van Nostrand Treinhold, New York. pp. 115-132. Singleton, P. (1995). Bacteria in Biology, Biotechnology and Medicine, 4th edition, John Wiley and Sons Ltd., New York, pp 232-266. Solabi, G. A. (1193). Industrial processing of soybean in Nigeria: A dream turns to reality. Keynote address presented at the national workshop on small scale and industrial level processing of soybean organized by IDRC/11TA Soybean utilization project, Ibadan. pp 12.16. Stewart, F. S. and Beswick, T. S. (1977). Bacteriology, Virology and Immunity for Students of Medicine, 10th edition, English Language Book Society, London; pp 23-29.

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Eze, V. C et al: Continental J. Food Science and Technology 2: 37 - 43, 2008 Thomas, C. G. A. (1994). Medical Microbiology, University Press Cambridge, UK. Turner, P. D. (1983). The importance of lipolytic Microorganisms in degradation of oil palm products in Malaysia, Quality of palm oil product in Malaysia, 2:52-62. Received for Publication: 08/01/2008 Accepted for Publication: 13/09/2008 Corresponding Author Eze, V. C Present Address: Department of Microbiology, Michael Okpara University of Agriculture Umudike, P. M. B. 7267, Umuahia, Abia State, Nigeria. E-mail: vin13eze@yahoo.com

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Continental J. Food Science and Technology 2: 44 - 49, 2008 © Wilolud Online Journals, 2008.

NUTRITIONAL AND ANTINUTRITIONAL VALUES OF ARMYWORMS (NOCTUIDAE SPODOPTERA SPP)

1Abulude, F.O, 1Ogunkoya M.O, 2Ogunleye R.O, 3Kayode, B.O and 4Osundare, F.O 1Department of General Studies, 3Department of Crop Production, 4Department of Extension and

Management, Federal College of Agriculture, Akure, Ondo State 2Department of Zoology, University of Ado Ekiti, Ado Ekiti, Ekiti State

ABSTRACT The nutrition and antinutrition of Noctuidae spodoptera spp were examined using standard methods of analyses. From the results, it was found out that protein content of the caterpillar was high (mean value, 24.30 %), while fat, NFE, moisture, ash and fibre ranged thus mean value (%): 1.22, 48.33, 22.70, 1.88 and 1.58. Sodium (662.20 mgkg-1), Calcium (415.25 mgkg-1) and Magnessium (345.28 mgkg-1) were relatively high. Nickel (2.42 mgkg-1) was the least mineral present. The results also showed that the sample consist on antinutrients. It would be recommended that adequate processing should be ensured before consumption. KEYWORDS: Nutrition, Noctuidae spodoptera spp, livestock, alternative source of protein, armyworms

INTRODUCTION Armyworms are the caterpillars of various Noctiudae, mostly spodoptera spp, which under certain condition of high population density behave gregariously, swarms will match from field to field devastatingly, defoliating entire crops, most species are migratory as adults (Akinseye 2007) The plants they attack are mostly Gramineae (Cereal and grasses), but the genus spodoptera is recorded feeding on plants from 40 different families containing at least 87 species of economic importance. Armyworms pass the winter as a partially grown in the soil or under debris in grass area. Activity and growth are continuous except during very cold weather. When fully grown, they stop feeding for four days, then pupate over a 15-20 day period, Adult energy in May and June, mating takes place at night during the 5th hour after sunset (Pyke, 1979) multiple matings usually occur. Females feed for 7-10 days on honey, nectar or decaying fruit before laying eggs. Eggs are laid at night in clusters of 25-134 on grass and on small grain leaves. A single female may live as adult for days and produce up to 2000 eggs hatch in 6-10 days. Armyworm caterpillars will vary in length form 2mm at first instance, larvae have strip as running the length of their body, one strip is present on each side and another stripe runs down the middle of the back. The nutritional needs of most of the world population especially the developing nations remain unsatisfied. This has spurred on various researchers into studying the nutrient composition of both conventional and non conventional dietary items (Abdullahi, 2000). Improving the quality and quantity of livestock feeds have also been main concern of both researches and farms, hence efforts have been geared towards the utilization of lesser-known and under-utilized animals which are indigenous to Africa. From this angle, we have thought it fit to establish the nutritional status of the armyworms with a view of recommending their use as an alternative source of protein. It is therefore the aim of this study, is to quantify the proximate, mineral, oxalate, tannins and phytate compositions of armyworm (Noctuidae spodoptera spp). Further study is on to quantify fatty acid, amino acid and multienzyme digestibility of this sample.

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Abulude, F.O et al: Continental J. Food Science and Technology 2: 44 - 49, 2008

Plate 1: Noctuidae spodoptera spp samples used for the analyses Table 1: Proximate composition (%) of the samples analyzed (n=3) Parameter Min Max Mean Standard Deviation Coefficient of variation (%)

Protein 21.30 25.20 24.30 1.27 5.23 Fat 1.00 1.35 1.22 0.55 45.08 NFE 48. 15 48.42 48.33 1.62 3.35 Moisture 22.10 23.00 22.70 0.71 3.13 Ash 1.52 2.10 1.88 0.50 26.60 Fibre 1.42 1.62 1.58 0.50 31.65

n= number of determinations, NFE= Nitrogen free extract Table 2: Mineral composition (mgkg-1) of the samples (n=3) Parameter Min Max Mean Standard Deviation Coefficient of variation (%)

Na 650.70 667.20 662.20 154.53 23.34 Fe 28.30 30.50 29.00 2.53 8.72 Mn 22.35 33.25 26.32 1.27 4.83 K 38.32 51.00 48.32 2.50 5.17 Zn 40.10 52.33 48.22 1.32 2.73 Cu 4.80 6.22 5.57 0.74 13.29 Ca 315.01 428.22 415.25 15. 22 3.67 Ni 1.85 2.62 2.42 0.72 29.75 Mg 325.28 401.35 345.28 6.82 1.98

Table 3: Antinutritional contents of samples analyzed (n=3)

MATERIALS AND METHODS Sample preparation. Armyworms (Noctuidae spodoptera spp) (Plate 1) used for this study was obtained at Federal College of Agriculture, Akure, Ondo State, Nigeria. The samples were collected between July and August 2007.

Parameter Min Max Mean Standard Deviation Coefficient of variation (%) (mgg-1) Tannins 50.25 72.10 60.35 2.70 4.47 Phytate P 1.32 2.25 1.87 0.50 26.74 Phytate 2.06 3.51 2.32 1.00 43.10 Oxalate 0.62 1.06 0.75 0.50 66.67

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Abulude, F.O et al: Continental J. Food Science and Technology 2: 44 - 49, 2008 They were washed in distilled water, sun dried for 10 days, ground in a Kenwood blender, sieved with 2mm wire mesh and store in an air-tight container at an ambient temperature prior to analyses. Mineral Composition. Two grams of powdered sample was in a muffle furnace (5500C) for 3h, dissolved in 2M HCL, filtered and made up to 50cm3. Minerals were read using a Pye Unican SP 9 spectrophotometer. Proximate Composition The ash, fiber and nitrogen free extract (NFE) compositions of the sample were determined using standard methods of AOAC (1990), protein content was determined using the micro-kjeldahl method. The percentage nitrogen was multiplied by 6.25 to obtain percentage protein. Antinutritional Composition Tannin Tannin was determined by the quantitative method of Markkar and Goodchild (1996). Phytate. The procedure of Young and Greaves (1940) as modified by Abulude (2001) was used for extraction, precipitation and determination of phytate. Oxalate The procedure proffered by Day and Underwood (1986) was employed for the above determination. All determinations were in triplicate. Results were statistically analyzed using mean, standard deviation and coefficient of variation in percent. RESULTS AND DISCUSSION The proximate compositions of the sample are shown in Table 1. All the determinations were on dry matter basis. The protein value was relatively high with mean value of 24.30% (±SD=1.27.CV%=5.23). The fat content was low ranging from 1.00-1.35%. The ash content was also low. Moisture and NFE were generally high in the sample. The results obtained for the protein content in the study were lower than those recorded for Cuban Boa (60.20-66.80%) Ogunkoya et al 2006), Larva (66.09%; Ekpo & Onigbinde, 2005) and crab (28.69-87.57%; Adeyeye, 2002), but higher than 10.50-17.50% recorded for Bouillon cube (Akpanyung, 2005) and tree barks (3.65-14.18%; Abulude et al 2004). The level of protein may contribute substantially to the total, daily intake of proteins by humans and livestock. The low fat content may not contribute to the flavour if roasted or dried. The fat content obtained in this study is fat lower than the amount found in most conventional foods like beef, chicken, egg, herring, mackerel and milk (Pyke 1979). This implies that a 100g sample of the armyworm may not meet the caloric fat needs of humans and animals and would not allow it to contribute significantly as a source of non-visible oil to any diet in which it may be present. The protein content would be very appropriate weaning food for young ones since the fibre content was low (1.42-1.62%), which means the protein may be liable to easy digestion. Where the fibre is relatively high, it has nutritional advantage as it will assist in reducing constipation and other attendant problems (Adeyeye 2000). The moisture content was in the range covered by Olaofe et al (1998) for grasshopper and Abdullahi (2000) for fish, but higher than those reported for mushroom samples (Esieet and Kayode, 2007). The relatively high moisture will not assist in keeping quality since the sample will go bad in time. Mineral contents (mgkg-1) of armyworm are presented in Table 2. The mean values ranged thus Na(662.20), Na(29.00) Mn(26.32), K(48.32), Zn(48.22), Cu(5.57), Ca(415.25),Ni(2.42) and Mg(345.28).

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Abulude, F.O et al: Continental J. Food Science and Technology 2: 44 - 49, 2008 These values compared with results obtained from the literatures cited. Elements both micro and macro have been reported to be useful to animals and humans (Abulude et al 2007). Insects are known to be rich sources of various macro and trace elements. These elements are probably accumulated for future use in adult exoskeletal and connective tissue synthesis (Ekpo and Onigbinde (2005). Sodium is an extra cellular cation involved in the regulation of plasma volume, acid-base balance, nerve and muscle contraction, High dietary sodium has been associated with essential hypertention (Lathan 1997), Iron is an important trace element in the human body. It plays crucial roles in haemopoiesis, control of infection and cell mediated immunity (Bhaskaram 2001). The deficiency of iron has been described as the most prevalent nutritional deficiency, and non deficiency anemia is estimated to affect more than one billion people worldwide (Trowbridge and Martorelli, 2002). The consequences of iron deficiency include reduce work capacity, impaired body temperature and regulation, impairment in behaviour and intellectual performance, and decreased resistance to infections (Dixon et al, 2004). Zinc is present in all tissues of the body (i.e both humans and animals) and is a component of more than 50 enzymes (Adeyeye 2000). Meat is the richest source of zinc in the diet and supplies one third to one-half of the total zinc intake of meat eaters. Zinc dietary deficiency has been found in adolescent boys. An estimated 20% of the world population is reported to be at risk of inadequate zinc intake (Hotz and Brown 2004). In Nigeria study has shown that zinc deficiency affects 20% of children less than five years, 28.1% of mothers and 43.9% of pregnant women (Dixon et al 2004). The sample appeared to be a good source of magnesium, sodium and potassium. Magnessium is an activator of many enzyme systems and maintains the electrical potential in nerves (Shils, 1973). Potassium is primarily an intracellular cation, in large part this cation is bound to protein and with sodium influences osmotic pressure and contributes to normal pH equilibrium. Plants and animals tissues are with sources of potassium this dietary lack is seldom found. Sodium is widely distributed in foods with the plants containing less than animal tissues. CONCLUSION The study shows that armyworms contain substantial amount of protein, carbohydrate and moisture which may contribute to the daily intake of these nutrients. The low levels of the antinutrients may be advantageous because they may not hinder the availability of the essential nutrients to consumers. However it is recommended that proper processing method should be put in place or used for the antinutrients to be destroyed or removed. REFERENCES Abdullahi S.A. (2000): Nutrient composition of three species of mormyrids in the Nigerian fresh water Nig. J. Tech. Educ. 17(1& 2): 177-183. Abulude F.O., Adesanya W.O., Ogunkoya M.O., Elemide O.A. and Esiet E.E. (2007): Nutritional composition of Ogi and its by-product. Acta Alimentaria. 36(4): 489-493. Abulude F.O., Onibon V.O. and Oluwatoba F (2004): Nutritional and anti-nutritional composition of some tree barks Nig. J. Basic and Appl. Sci. 13:43-49. Abulude F.O. (2001): Mineral and phytate content of vegetable grown in Nigeria and calculation of their phytate: Zn and Ca: phytate molar ratio. Adv. Food Sci. 23(1): 366-39. Adeyeye E.I. (2002): Determination of the chemical composition of the nutritionally valueable parts of male and female West African fresh water crab Sudananantes africanus. Int. J. Food Sci. & Nutr. 53:189-196.

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Abulude, F.O et al: Continental J. Food Science and Technology 2: 44 - 49, 2008 Adeyeye E.I. (2000): Determination of the elemental composition of the nutritionally valuable parts of male and female common West African fresh water crab sudananautes africanus. Int. J. Food Sci & Nutr. 47: 111-116. Akinseye A. S. (2007): The proximate, elemental, oxalate, phytate and tannin compositiond of armyworms (Noctuidae spodoptera spp). National Diploma Thesis, Federal College of Agricultre, Akure, Ondo State, Nigeria. Akpanyung E.O. (2005): proximate and mineral element composition of bouillon cubes produced in Nigeria. Pakistan J. Nutr. 4(5): 327-329. AOAC (1995): official methods of Analysis. 5th edition, Washington D.C. Association of Official Analytical Chemists pg 6-16. Bhaskaram P (2001): Immunobiology of mild nutrient deficiencies. Br. J. Nutr. 85:s75-s80. Dixon B.M., Akinyele I. O, Oguntona S. Nokue R.A., Sanusi and Harris E.M (2004): Nigeria food consumption and nutrition survey 2001-2003 summary IITA, Ibadan, Ekpo K. E and Onigbide A.O (2005); Nutritional potentials of larva of Rhynochoplorus phoenicis (F) Pakistan J. Nutr. 4(5): 287-290. Esiet E. E. and Kayode B. O. (2007): Proximate composition and economic feasibility of some mushroom consumed in southwestern Nigeria. Cont. J. Food Sci & Tech. 1(3): 7-10. Hotz C and Brown K.H (2004): International zinc nutrition consultative group (IZINCG). Technical Document No 1. Assessment of the risk of zinc deficiency in populations and options for its control. Food and Nutrition Bull. 25:s130-s162. Latham M.C. (1997): Human nutrition in the developing world. FAO Foods and Nutrition Ser. No 29, Rome. Makker M and Goodchild G (1996): The similarity between oxalic present in leaf and oxalate present in leaf and oxalate present in rat. Br. J. Nutr. 39(2): 233-414. Ogunkoya M.O., Abulude F.O. and Oni A.B (2006): Determination of anatomical, proximate, minerals, oxalate, tannin and phytate compositions of Cuban Boa (Epicrates anquifer). Electr. J. Environ, Agric., and Food Chem. 5 (1):1661-1166. Olaofe O. Arogundade L. A., Adeyeye E.I and Falusi O.N (1998): Composition and food properties of the variegated grasshopper, Zonocerus variegatus. Trop. Sci. 36:233-237. Pyke M. (1979): The science of nutrition. In: Science in nutrition. John Murray (Publishers) Ltd, London pp 251-258. Shils M.E. (1973): Magnesium. In modern nutrition in health and disease, eds RS Goodhart and M. E.Shils, Ch 6, Sect. B. Philadelphia P. A: Lea and Ferbiger. Trowbridge F and Martorell R (2002): Forgoing effective strategies to combat iron deficiency. Summary and Recommendations. J. Nutr. 85: s75-s80. Young S.M. and Greaves J. S. (1940): Influence of variety and treatment of phytin content of wheat. Food Res. 5:103-5.

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Abulude, F.O et al: Continental J. Food Science and Technology 2: 44 - 49, 2008 Received for Publication: 08/01/2008 Accepted for Publication: 13/09/2008 Corresponding Author Abulude, F.O Department of General Studies, Federal College of Agriculture, Akure, Ondo State.