Academia Journal of Food Research 1(3): 050-058, September 2013 DOI: http://dx.doi.org/10.15413/ajfr.2012.0117 ISSN: 2315-7763 ©2013 Academia Publishing

Research Paper

Production and evaluation of weaning meal from fermented red maize fortified with cowpea

Accepted 16th June, 2013

ABSTRACT

A weaning food blend was prepared from an improved variety of maize (Marggi Red) and cowpea. Standard method was used for the analysis. The protein content Modu S, Ibrahim Z, Hajjagana L, of the weaning blend (12.68±0.45) was not able to meet the recommended Dietary Falmata AS, Babagana M and Allowance of infants of weaning age. The Calcium content of the weaning food Bintu BP blend (50.08±0.09) was compared favorably with commercial weaning food Department of Biochemistry, frisocrem® (52.00). Processing resulted in increase in-vitro protein digestibility Faculty of Science, University (77.91±0.29 to 83.40±1.96) at one hour and (83.93±0.12 to 87.86±0.05) at six of Maiduguri hours of the composite meal.

Corresponding author email: [email protected] Key words: red maize, cowpea, nutritional value.

INTRODUCTION

Weaning is a gradual process of introducing solid foods to food blends. an infant’s diet, alongside breast milk from the age of three to four months, since the breast feeding along cannot meet Maize yields more flour, with much less bran than wheat. the infant nutritional requirement (Salmon et al., 2008). In However, it lacks the protein gluten of wheat and therefore, Nigeria, traditional weaning foods consist of monocereal makes baked food poor rising capability (USDA Nutrient grains, prepared from either millet, maize or sorghum Database, 2009). referred to as “Ogi” or “Kamu” which is of poor nutritional Maize and cornmeal (ground dried maize) constitute a value (Hellstrom et al., 1981). The major problems staple food in many regimes of the world, introduced in to associated with infant during transitional phase of weaning Africa by Portuguese in the 16th century, maize has is the protein energy malnutrition (PEM), which is becomes African’s most important staple crop (FAO, 2009). associated with wasting condition resulting from a diet Maize meal is made in to a thick porridge in many cultures inadequate either protein or energy (calories) or both. This from polenta of Italy, the angu of Brazil, the mamaliga of can visualized as either Kwashiorkor or Marasmus (Pollitt, Romania, to mush in the USA or the food called mealic pap 1995). in South Africa and Sadza, nishima and ugali in other parts The supplement of these locally available cereals with of Africa. legumes, which are good sources of protein rich foods, will In Nigeria Maize is prepared and consumed in a give rise to weaning food that gives the infant enough multitude of ways vary from one ethnic group to the other, energy and nutrient for its nutritional requirements for instance, Maize grains are prepared by boiling or (Marero et al., 1988). roasting as paste (‘eko’), ‘aba do’ and elekute in some part of the country. Maize is an all-important crop which provides an avenue for making various types of foods. It is AIMS AND OBJECTIVE OF THE STUDY also used as a raw material in many industries (Osagie et al., 1998). (i) To produce a weaning food from fermented red maize, ‘Tuwo’ (Yoruba), ‘tuwo-masara’ (Hausa), ‘Oko’ (Egun) etc fortified cowpea that will be adequate to meet the is a very important and popular stable food among various nutritional requirement of infants during weaning period. ethnic groups in Nigeria from Maize. ‘Maasa’ or Wainna are thick porridges while ‘Maasa’ is small size, Wainna is big (ii) To evaluate the chemical composition, mineral element eating with sugar sprinkled on it or with pumking soup are level and in in-vitro protein digestibility of the weaning all prepared from maize in Nigeria, Wainna cakes also be Academia Journal of Food Research; Modu et al. 051

made with mixture of cassava flour (Manihot esculenta) and (Scotland), Muffle furnace, beakers, burretes, crucibles, millet flour (Lancaster et al., 1982). Other uses of Maize Soxhlet extractor, round bottom flask, digesting tube, includes ‘cous cous’, Akple, Ukejuka, Uwate, Nakia, Dambu, distilled water, indicator (methyl orange and bromcresol Alubosa, Abari and Egbo are produces from maize cereal green),pipettes, conical flask, distiller, Petri dishes, grains that can either be cooked, roasted, fried ground, measuring cylinder, funnels, pair of tongs. pounded or crushed to prepared in to this various food items (Abdulrahaman et al., 1997). Most importantly, cowpea seed is nutrition’s component List of chemicals used for in vitro protein digestibility in the human diet, as well as a nutritious livestock – feed green manure and cover crop for maintenance of Phosphate buffer (pH 7.5), 11% trypsin, 0.1N HCl, productivity of soils. The dried seeds may be grounded in to neutralized formalin, filter paper, test tubes. meal or flour, which is used in a number of ways. The fresh seeds and immature pods are eaten as vegetables. They may be frozen or canned. The young shoots and leaves are List of material used for mineral element analysis eaten as spinash and provide one of the most widely use pot herbs in tropical Africa (Sinha et al., 1999). Muffle furnace, beaker, distilled water, Whitman filter In Nigeria cowpea (Vigna unguiculata) are an important paper, atomic absorption spectrometer. grains legumes and source of protein in the diets. Cowpeas are used extensively to fortify cereal – based weaning foods (Uwaegbute, 1991). They are also processed in to paste or List of material used for titratable acidity (TA) flour and used as a food ingredient or starting material for a variety of local foods (Mc Watter, 1983). Phenolphthalein, 0.1N sodium hydroxide, beaker, restort The protein in cowpea seed is rich in the amino acids stand, 10 ml pipette, burette, pH meter. lysine and tryptophan, compared to cereals grains, however, it is deficient in methionine and cystein when compared to animal proteins. Therefore, cowpea seeds is Collection of sample valued as a nutritional supplement to cereals and an extender of animals. The nutrient content of mature Red maize Zea mays L, (Marggi Red) and cowpea Vigna cowpea seed is estimated as protein 24.8%, fat 1.9%, fiber Unguilata (Biu Local) were purchased at Maiduguri Monday 6.3%, carbohydrate 63.6%, thiamine 0.00074%, Riboflavin Market, Nigeria and identify by Dr. O, Oluwana, Head of 0.000042% and Niacin 0.00281% (Singh, 2003). cereals grains from Lake Chad Research Institute (LCRI), Legumes such as cowpea are a particular rich source of Maiduguri, Nigeria. natural anti oxidant such as protease inhibitor, Amylase inhibitor, flatus factor, saponins, cynogenic glycoside etc (Singh, 2003). Preparation of the Kamu from red maize Although, the natural lactic acid fermentation improved the contents of certain B-group vitamins considerable The ‘Kamu’ (Ogi) was prepared (Figure 1) by the method variations occurs depending on the nature of the raw described by Akingbala et al. (1981). One hundred grams materials, microflora, temperature and the nature for the (100 g) of clean Red Maize (raw grain) were soaked in method of determination for these vitamins. water for one hour to get rid of foreign matter and damage The microbial fermentation of cereals and cereal-legume grains and steeped in 200 cm3 of distilled water (1:2 ratios) bleeds improves their relative nutritive value, availability of for 72 h (3 days) at the end of the 0, 28, 48 and 72 h the pH protein, amino acids (lysine, threonine, tryptophan, and and titratable acidity was taken, and finally the top water methionine), carbohydrates, contains B group vitamins and was decanted and 200 cm3 of distilled water was added and minerals. The fermentation of cereals and legumes milled with warring blender for 4 min at rheostate setting decreases or eliminate certain antinutrient like phytates, of 120. The slurry obtain was served through nylon cloth to protease inhibitors, flatus factors and lectins. separate bran after which the water was decanted and the Kamu was sun dried to a constant weight.

MATERIALS AND METHODS Preparation of the cowpea (Biu local) List of chemicals used for proximate analysis One hundred grams (100 g) of the cowpea was cleaned of Sulphuric acid, Kjedahl tablet, sodium hydroxide, boric acid, dirt and soaked in distilled water for 20 min. The cowpea hydrochloric acid, petroleum ether, trichloroacetic acid, was then diluted using a pestle and mortar then washed to glycylacetic acid, nitric acid, filter paper, Monak desiccator separate the husk, after which it was dried to constant Academia Journal of Food Research; Modu et al. 052

Raw materials

Clean

Red maize Cowpea

Soaking Soaking

Dehulling Fermentation (72hours)

Drying

Milling

Roasting Drying

Grinding

Weaning food ogi

Store Store

Figure 1. The flow chart of ogi preparation. Sourer: Pak. J. Nutr., 8(10): 1671-1675, 2009.

weight and roasted. The cowpea was then ground in to a involves gyratory movement of grains in relation to sieve find powder. mesh, at the same time a reciprocating up and down movement is performed on the nest of sieves so that the action jolts the grains at all possible angles. The shaker Preparation of the weaning food blend generally hold up to nine or ten sieves including a collecting pan and lid set under vibration until a sufficient grains has The blending of the weaning meal was done in the ratio of been sieved then the process stop and determined the 70:30 as described by Akapunam (1984) that is 70 parts of various sizes of grains retained. fermented Red Maize (Kamu) and 30 parts of milled cowpea were mixed together by using blender. Hardness of grain

Sieving analysis The degree of hardness of grains is obtained by using Ward’s Natural science establishment Inc. Rodester New The sieve analysis was obtained using British Standard York method by comparing with that of ten standards, nine sieves, wire mesh series (B.S 410: 1969). This British of which are included in this set. A given grains would, for Standard provide a wide range of fine mesh size ranging example, be described as having “hardness number 4” or by from 16 mm to a minimum of 38 microns. In this process it those of the scale, remembering that a harder grains will Academia Journal of Food Research; Modu et al. 053

cause a scratch mark an softer one only. These standards Ash determination are Talc, Selenite, Calcite, Fluorite, Apalite, Microline, Quartz, Topaz, Diamond. Ash was determined by the method of AOAC (2002). Two grams of each sample was added in separate crucible and placed in a muffle furnace and then ashed at 560°C for three pH measurement (using pH meter) hours. The sample were removed from the furnace, cooled in desiccator to room temperature and weighted. pH measurement was done using share analytical method Percentage ash was calculated using the formula below: of food analysis (2010), by placing the sample in to clean beaker and dip the electrode of pH meter until the reading Wt of crucible + Ash –Wt of empty crucible become stable then the value is read. % Ash= ×100 Initial Wt of sample

Measurement of titratable acidity (TA) Fat determination Titratable acidity was measured using share analytical method of food analysis (2010), by pipetting 10 ml of the The ether extract was determined using Soxhlet apparatus. sample in to a 50 ml beaker, add 3 drops of phenolphthalein Two grams of the sample was weighed in to a thimble and indicator, filled the burette with 0.1 ml NaOH standard 200 ml of petroleum either was measured into a conical solution. Taking note of the initial reading, titration was flask. The solution was heated at 45°C at 1 h interval for 2 h. done by slowly rotating the burette stop cock. The beaker The flask was removed, reweighed and percentage fat was swirled so that the sample and NaOH would mix very sample is determined using the formular: well until the solution become pin for 30 s then the final burette reading was read to an accuracy of 0.1 ml. To Wt of fat calculate the volume required for the titration, the initial %fat = × 100 reading was subtracted from the final reading. Wt of sample It is computed as follows:

V (N) (meq.wt) (100) TA (g/100 ml) = Crude protein determination (1000)(V) Crude protein content was analyzed using Where: V = is volume of sodium hydroxide used, N = Kjeldahl procedure, 2 g of samples was weighed into a Normality, Meq.wet = is milliequivalent weight of standard, digestion tube and 2 tublets of Kjeldahl were added, 2 ml of V = is sample volume. concentrated sulphuric acid (conc. H2S04) was added, on to the tube and digested at 420°C for 3 h. After cooling 80 ml of distilled was added into digested solution. About 50 ml of Proximate analysis 40% caustic soda (NaOH) was added on to 50 ml of digested solution and then placed on heating section of the The proximate composition is determined by AOAC (2000) distillation chamber, 30 ml of 4% boric acid, plus method. bromocresol green and methyl red indicator was put into conical flask and placed underneath the distillation chamber for collection of ammonia, the solution changed Determination of moisture from orange to green colour. About 0.1 normal solution of HCl was weighed in to burette. The conical flask containing The direct oven method (AOAC, 2000) was used for this the solution was titrated until the colour change from green analysis. Ten grams of each sample was weighed using to pink. The burette reading was taken. The crude protein electric balance and placed in a Petri dish of known weight. was calculated using the formular: The sample was then allowed to dry in the air oven set at 108°C for 8 h. The dish and sample were cooled in a desicators and then weighed. This procedure was repeated until a constant weight was obtained. The moisture content of each sample was calculated as follows: Where: A = Ml of acid used for titrating sample, B = Ml of Wt of orig. sample –Wt of orig. dried sample acid used for titrating blank, N = Normality of acid used for % moisture= ×100 Wt of the orig. sample titration, 100 = conversion to percentage. Academia Journal of Food Research; Modu et al. 054

Crude fiber determination Mineral element analysis

Two grams of the sample was placed in a 450 ml conical Twenty grams of each sample was weighed using electric flask and 50 ml of trichloracetic acid reagent (TCA) was weighing machine. The sample was then ashed in a furnace added, the mixture was boiled and refluxed for 40 min. The at arshing temperature of 550°C. After the arshing, it was flask was removed and cooled to room temperature. Filter weighed to be 2 g out of which 1 g of ashed sample was paper was used to filter the residue. The residue obtained digested. One gram of the sample was in a 200 ml beaker was 4 times with hot water and once with petroleum ether and 30 ml of nitric acid and distilled water was then added then the filter paper plus the sample were folded together to sample in the beaker. The sample is then wormed over and dried at 30-60°C in an oven for 24 h, reweighed then water bath for 35 min and then allowed to cool. The washed at 610° and reweighed flask was removed, digested sample is then filtered using white man filter reweighed and percentage fat sample is determined using paper and diluted with water to volume of 100 ml. the formular: The sample was then run at a particular wavelength using Atomic absorption spectrometer (model AA analyst 400) of Wt of fiber Perkin Elmer Company product to determine the various %fiber = ×100 minerals element present in the samples. Wt of sample

Water absorption capacity (WAC) Determination of carbohydrate Water absorption capacity was determined by the method This was determined by differences: of Celgla et al. (1977) with slight medications, 10 g of each sample were weighed in a 100 ml beaker. A known volume % carbohydrate = 100 – (% moisture + Ash + % protein + (10 ml) of water was pipette in to the beaker, carefully % crude fiber). stirred and allowed to equilibrate for hour at room temperature (23 – 25°C). After complete water absorption, the sample was further treated with 0.01 ml water portion, Determination of energy value at 10 min interval before usual observation. The volume that gave a complete absorption of water (no visible free This was obtained using the atwater factors (physiological water) is compute as: fuel values) of 4kcal, 4kcal and Gkcal per gram of carbohydrate, protein and fat content of the samples WAC = Initial weight – Final weight (after absorption of respectively. water).

Determination in-vitro protein digestibility (Nills, RESULTS 1979) Physical characteristics One milliliter of 11% trypsin was introduced in to 5 test tubes. Four milliliter of phosphate buffer of pH 7.5 was Table 1 present the grain diameter (mm) for maize at added to each test tube 1 ml of 0.1 N HCl was added and different mesh sizes range between 0.00 – 28.92 with mesh allowed to stand to equilibrate 1 ml of 1% ‘Kamu’ was size 6.70 having the highest grain size of 28.92 and also the added to all the rest tubes (labeled as digestibility at 1 and cowpea exhibited a similar trend of grain diameter range 6 h). The reaction in each of the test tube was stopped with 0.00 – 27.36 and mesh size 6.70 having highest diameter of 5 ml of neutralized formation at 60 min and 6 h. The 27.36. Table 2 show a hardness number of 5- apotile while content of the test tubes were then filtered using filter cowpea show 4- flourite. paper. The filter papers were dried in an oven at 108°C for Table 3 show the pH and titratable acidity of “Kamu” 3 h. from maize. A decrease in pH with an increase in titratable The Nitrogen of the undigested sample was determined by acidity was exhibited during the fermentation process. the Kjedahl method. Table 4 shows the proximate composition of the raw and process maize and cowpea. There is no significant CP1-CP2 difference in the moisture content of the raw and processed % in vitro protein digestibility = ×100 maize. The protein and fat content of the processed maize is CP1 significantly higher than that of the raw maize and thus the moisture content of the cowpea showed a significant Where: CP1 = total protein of unprocessed grain, CP2 = difference. While the Ash, shows a significant decreased. Total protein after digestion with trypsin. Table 5 present the proximate composition of the Academia Journal of Food Research; Modu et al. 055

Table 1. Sieve analysis. 28.92 at mesh size 6.70 of the maize as well as the cowpea exhibited a similar mesh size in diameter of 6.70 retend the Grains diameter (mm) highest grain of 22.36, while the maize show a hardness Mesh (mm) Maize Cowpea number of 5-apotile and the cowpea show 4-flourite in 16 0.00 0.00 Table 2. 11.20 0.00 0.00 8.00 11.38 1.45 6.70 28.92 27.36 pH and titratable acidity (TA) 4.10 9.59 21.17 A decrease in the pH (Table 3) with an increase in titratable 3.35 0.00 0.01 acidity was exhibited during the fermentation process, and 2.00 0.00 0.01 this is similar to the report of (Akinrele, 1970). The acidic 1.00 0.00 0.00 nature of the product could be due to the production of lactic acid produced by microorganism associated with maize fermentation (Newman et al., 1984). Table 2. Grains hardness (N).

Grains Hardness number Proximate composition Maize 5 – Apotite Cowpea 4 – Fluorite The proximate composition (Table 4) shows that there is no significant difference in the moisture and ash content between the raw and the processed maize. The protein of Table 3. pH and titratable acidity (TA). the processed is significantly higher than that of the raw

maize this could be as a result of improvement in the Time pH Titratable acidity protein content of the maize during fermentation. This is 0 6.10 0.000031 similar to the report of Nanson and Field (1986). The 24 4.50 0.000062 improvement to the fat content of the processed may be 48 4.70 0.000112 due to increase in activity of lipolytic enzymes in the 72 4.30 0.00016 fermentation medium which hydrolyzes fat to glycerol and fatty acids. The free fatty acids are used by the fermenting organisms for the synthesis of new lipids (Ezeokenkwo, weaning food blend compared with commercial weaning 2004). The moisture content of the cowpea shows a food frisocrem. The moisture content compared favourably significant decreased in the processed cowpea. This and the protein of the blend does not meet the protein attribute longer shelve life span to the processed cowpea content of the frisocrem. than the raw cowpea. Table 6 presents the mineral element composition of the The protein content of the weaning blend (Table 5) is raw and processed maize and cowpea. There is significance compared low with respect to the commercial weaning food increase in the level of Ca, Na, P, and Fe in the processed frisocrem®, though it provide one third of the RDA (14%) maize while there was a significant decrease in the level of as reported by WHO (FAO/WHO/UNU, 1985) and National ca, Mg, Na, and P in the processed cowpea. Institute of Nutrition (ICDS, 1992) for children and rural Table 7 shows the mineral element composition of the mothers, while fiber and Ash content was not detected. weaning food blend compared with commercial weaning food frisocrem with a Ca level compare favourably and the Na, P and Fe of the blend does not meet the RDA of the Mineral element composition frisocrem®. Table 8 shows the in vitro protein digestibility of raw and The increased in the content of Ca, Na, P and Fe in the processed maize and cowpea with a significantly increase processed maize could be as a result of improvement in the in the level of both processed maize and cowpea. mineral content of maize during fermentation (Reddy and Salunkhe, 1980). The increase in the level of Fe could be due to the reduction of phytic acid during fermentation DISCUSSION since lactic acid fermentation changes a diet of low iron bioavailability (Suanberg and Sandberg, 1988). The Physical characteristics decreased in the content of ca, Mg, Na and P of the processed cowpea could be attributed to the loss in ash The sieve analysis of the grains (Table 1), in diameter (mm) content during dehulling of cowpea (Akingbala et al., 1981). at different mesh sizes ranged between the highest grain of The blend ratio of the weaning food (Table 7) shows that Academia Journal of Food Research; Modu et al. 056

Table 4. Proximate composition of raw and processed maize and cowpea.

Maize Cowpea % Raw Processed Raw Processed Moisture 4.10 ± 0.17a 3.80 ± 0.03a 7.10 ± 0.03c 2.50 ± 0.05d Ash 1.50 ± 0.56a 1.50 ± 0.03a 2.33 ± 0.45c 1.50 ± 0.01d Protein 6.98 ± 0.12a 8.74 ± 0.02b 9.54 ± 0.12c 11.99 ± 0.04d Fat 5.00 ± 0.20a 6.00 ± 0.02b 12.00 ± 0.36c 13.00 ± 0.02c Fiber 6.00 ± 0.22a 8.48 ± 0.05b 7.00 ± 0.18c 8.00 ± 0.07d Carbohydrate 82.50 ± 2.40a 71.46 ± 0.05b 69.49 ± 0.02c 65.51 ± 0.49d Energy (kcal) 403.00 ± 1.49a 365.88± 0.36b 424.00±2.90c 427.01±0.08c

Values are recorded as mean ± SD of three determination, values in the same row with difference superscript are significantly different (P < 0.05).

Table 5. Proximate composition of weaning blend compared with commercial weaning food frisocrem®.

Maize/cowpea Commercial weaning food % (70:30) mg/100 g Frisocrem (rice) mg/100 g Moisture 1.90 ± 0.02 2.0 Ash 1.00 ± 0.02 ND Protein 12.68 ± 0.09 16.00 Fat 13.33 ± 0.45 ND Fiber 12.33 ± 0.18 13.20 Carbohydrate 63.79 ± 0.10 65.10 Energy (kcal) 414.00 ± 0.27 455

70 parts of processed maize to part of cowpea, ND – Not detected, Value recorded as mean ± SD of three determinations.

Table 6. Mineral composition of raw and processed maize and cowpea.

Maize Cowpea Mineral element Raw Process Raw Process Ca 311.53 ± 2.31a 366.27 ± 0.40b 384.66 ± 0.03c 216.33 ± 0.02d Mg 153.39 ± 1.32a 149.42 ± 0.32b 224.06 ± 0.02c 113.61 ± 0.02d Na 139.75 ± 0.53a 225.60 ± 0.02b 127.20 ± 0.03c 119.55 ± 0.02d P 128.50 ± 0.09a 212.70 ± 0.03b 209.48 ± 0.06c 151.20 ± 0.03d Fe 4.45 ± 0.03a 5.98 ± 0.06b 488.00 ± 0.02c 5.03 ± 0.01d K 37.50 ± 0.03a 22.70 ± 0.02b 29.70 ± 0.06c 19.05 ± 0.03d Zn 13.55 ± 0.05a 12.46 ± 0.01b 10.22 ± 0.03c 13.72 ± 0.02d

Values recorded as mean ± SD of three determination, Values in the same row with different superscript are significantly different (P <0.05).

the Ca content compared favorably with the commercial in level of anti nutrient (Scheinfeld and Mokashi, 2001). The weaning food frisocrem® and the level of Na, Fe, P and Zn heat treatment of cowpea also helps in increasing the is compared closer, while Mg is not detected. digestibility of the weaning food (C.A.C 1999).

In vitro protein digestibility Conclusion

The processing process helps in increasing the in vitro The values of the proximate and the elemental analysis protein digestibility significantly, and this is due to does not meet the recommended dietary allowance of Academia Journal of Food Research; Modu et al. 057

Table 7. Minerals element of weaning blend compared with commercial weaning food.

Mineral element Maize/cowpea 70:30 (mg/100 g) Commercial weaning food (frisocrem (rice) mg/100 g) Ca 50.08 ± 0.09 52 Mg 386.48 ± 1.40 ND Na 598.77 ± 1.40 665 P 398.39 ± 1.03 425 Fe 6.90 ± 0.09 9 K 450.30 ± 1.01 500 Zn 149.86 ± 0.22 180

Value is recorded as mean ± SD of three determinations, 70 parts of processed maize to 30 parts of cowpea, ND- not detected.

Table 8. In vitro protein digestibility of raw and processed maize and cowpea.

Maize Cowpea Raw Processed Raw Processed Digestibility at 1 h 77.99 ± 0.29a 83.40 ±1.96b 89.51 ± 0.21c 93.32 ± 0.38d Digestibility at 6 h 83.93 ± 0.12a 87.86 ± 0.05b 90.56 ± 0.16c 96.66 ± 0.12d

Values are recorded as mean ± SD of three determination, value in the same raw with different superscript are significantly different (P<0.05).

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