CentralBringing Excellence in Open Access

Journal of Human Nutrition & Food Science

Cite this article: Oche I, Patrice-Anthony O, Ubwa S, Adoga S, Inegedu A, et al. (2017) Micronutrients, Oxalate and Phytate Composition of a Potential Infant Food Made from Soybean, Yellow Maize, Short Rice, Egg Yolk and Crayfish. J Hum Nutr Food Sci 5(3): 11156.

*Corresponding authorIkese Oche, Department of Chemistry, Benue State University Makurdi, Nigeria, Tel: 234 8032497326; Email:

Submitted: 15 September 2017

Accepted: 04 November 2017

Published: 07 November 2017

ISSN: 2333-6706

Copyright© 2017 Oche et al.

OPEN ACCESS

Research Article

Micronutrients, Oxalate and Phytate Composition of a Potential Infant Food Made from Soybean, Yellow Maize, Short Rice, Egg Yolk and CrayfishIkese Oche*, Okoye Patrice-Anthony, Simon Ubwa, Sunday Adoga, Audu Inegedu, Obande God’s will and Yvonne AkaasahDepartment of Chemistry, Benue State University Makurdi, Nigeria

Abstract

A novel infant food was formulated by beneficiating a cereal legume composite prepared from yellow maize, short rice and soybean using egg yolk, crayfish and banana flavour. Simultaneous equation and material balance methods were used to determine the blend proportions with which the cereal-legume composite was prepared. The mineral and vitamin compositions of the formulated food were determined by Atomic Absorption Spectrophotometer (AAS) and Reverse Phase High Performance Liquid Chromatography (RP-HPLC) respectively and then statistically compared with the mean compositions of same in two common proprietary formulae sold in the markets and in applicable cases with the recommended daily allowance for infants between 6-59 months. The micronutrient content of the formulated food compared quite favourably with those of the proprietary formulae and in most cases was even higher in the formulated food than in both proprietary formulae. With the exception of copper the micronutrient content per 100g of the formulated food compared adequately with their corresponding RDA’s and the formulated food will more than adequately supply the RDA’s for each mineral nutrient and vitamin in infants between 6-59 months of age if they are fed at least 150g of the formulated food daily. This makes the formulated food an excellent source of micronutrients for infants between 6-59 months as it is remarkably sufficient in terms of micronutrient composition. Oxalate and phytate in the formulated food were found to be 0.78 % and 1.48 % respectively.

Keywords•Infant food•Proprietary formulae•Vitamins•Minerals•Phytate•Oxalate

INTRODUCTIONInfant malnutrition is a major problem bedevilling

developing countries, and has been reported to be largely responsible for infant mortality [1]. This situation is further worsened by the high cost of proprietary infant formula which places them beyond the reach of low-income families who often revert to inadequately processed traditional foods that consist mainly of un-supplemented cereal porridges made from maize, sorghum and millet [2]. The above trend has triggered a number of frantic efforts towards the formulation of a wide range of infant food products. Also, It may be argued that whereas some of the formulations are scientific a portion of these still appear to be arbitrary and unscientific. The latter approach to food formulation is perhaps what has triggered a new dimension to the problem, where in some cases it would appear that less than sufficient premium is placed on the micronutrient adequacy of such formulations as well as their anti-nutrient levels especially as relates to their recommended daily allowances (RDAs) and maximum permissible limits in infant foods respectively.

There have been many attempts to develop infant food products from local feed materials [2-8] and several claims continue to be made by researchers and feed formulators regarding the nutritional adequacy of different food formulations. In some cases, if a food product meets the macronutrients requirements of target consumers, the formulation/food product is adjudged to be suitable even prior to a micronutrient-level assessment [5,9,10,11], yet these too play essential roles in optimum nutrition and the wellbeing of infants. This perhaps explains the myriad of micronutrient deficiency diseases in infants fed with some novel formulations. Therefore it becomes expedient to begin to verify all claims by subjecting new food formulations to micronutrient profiling assays that allow the formulators, food product regulatory agencies and consumers to truly verify the nutritional adequacy of potential new food formulations as a way to forestall infant malnutrition. This study seeks to determine the micronutrient, phytate and oxalate compositions of a potential infant food made from soybean, yellow maize, short rice, egg yolk and crayfish, with a view to

CentralBringing Excellence in Open Access

Oche et al. (2017)Email:

J Hum Nutr Food Sci 5(3): 1116 (2017) 2/6

ascertaining whether it compares favourably with two selected proprietary formulae sold in the market and the recommended daily allowances of the profiled micronutrients. It will also show if the oxalate and phytate levels in the potential food is within the maximum permissible limit in foods.

MATERIALS AND METHODS

Sample procurement and preparation of flours

Soybean, short rice and yellow maize grains were procured from local markets across Benue State, then separately composited according to variety and used for the infant food formulation and subsequent chemical analysis. Chicken eggs, crayfish, banana flavour and two proprietary infant formulae designated as formulae A and B were also procured. Flours were made from yellow maize (Zeaindentata) and short rice (Oryza japonica) as described in Scheme 1 and 2 respectively. Crayfish powder, Soybean flour and egg yolk powder were prepared as shown in Scheme 3, 4 and 5 respectively. The soybean, short rice and yellow maize flours are as shown in Figure 1.

Feed formulation

A cereal-Legume composite (CLC) was formulated by blending the flours produced according to equation (1) and thereafter, it was beneficiated according to equation (2) to give the formulated infant food (FIF).

CLC (100%) = 41.96% SR + 3.61% YM + 54.43% SF (1)

FIF (100%) = 92% CLC + 2.5% EY + 2.5% + CFh + 3% + 3% BF (2)

Where:

SR, YM, SF, EY, CFh and BF represent short rice flour, yellow maize flour, soybean flour, egg yolk powder, crayfish powder and banana flavour respectively.

The above blend proportions were mathematically derived by treating the protein and fat compositions in each of the three grain species used, as nutrient constrains in simultaneous equation and material mass balance equations as described in the formulation of the food by Ikeseet al. [12]. The formulated food, proprietary formulae A and B are shown in Figure 2, 3 and 4 respectively.

Food analysis

Mineral determination: Two grams of sample was ashed in a muffle furnace at 600°C. The ash obtained was digested on a hot plate in 10 mL of 5M HCl. The digested sample was carefully washed with deionised water and quantitatively transferred into a 50 mL volumetric flask before diluting to volume. The solution was then directly analyzed using Atomic Absorption Spectrophotometer (SOLAR 929 Unicam A.A. Spectrophotometer, UK) for the following minerals Na, Fe, Mg, Ca, K, Pb, Zn, Cu, Se, Cd and Ni at wavelengths of 589.0, 248.3, 285.2, 422.7, 766.5, 217.0, 213.9, 324.7, 196.0, 288.8 and 232.0 nm respectively.

Vitamin Determination: Water-soluble and fat-soluble vitamins were determined using reversed-phase HPLC (RP-HPLC) and non-aqueous reversed-phase HPLC (NARP-HPLC) respectively after water soluble vitamins and fat soluble

Figure 1 (L-R) Soybean, Short rice and Yellow maize flours.

Figure 2 Formulated Infant Food.

Yellow Maize Grain

Wash and air-dried

Steep in 2M Ca(OH)2 for 12hrs

Wash with distilled water

Malt for 96 hours

Wet mill and ferment for 48 hours

Sieve with 80µm mesh, lump, oven dry and mill

Yellow maize flour

Scheme 1 Preparation of Yellow Maize flour.

CentralBringing Excellence in Open Access

Oche et al. (2017)Email:

J Hum Nutr Food Sci 5(3): 1116 (2017) 3/6

Short Rice Grain

Sort, wash and steep in water for 10 mins

Parboil with minimal H2O for

Oven-dry at 105 0C for 4 hours

Mill

Short rice Flour

Scheme 2 Preparation of Short Rice Flour.

Scheme1: Preparation of Yellow Maize

Dried Crayfish

Clean by hand sorting, wash and soak

i f h

Strain and oven-dry for 30 mins

Steep in excess fresh lime juice

Strain and oven-dry for 1 hour

Mill

Sieve with 80 µm mesh

Crayfish Flour

Scheme 3 Preparation of Crayfish Flour.

Scheme 2: Preparation of Short Rice Flour

Soybean Grains

Clean and sort by manual handpicking

Wash and soak in water for 14 hours

Dehull and wash severally with water

Boil for 15 mins and air dry for 48 hours

Roast and Mill

Sieve with 80 µm mesh

Soybean Flour

Strain and blanch for 20 mins

Scheme 4 Preparation of Soybean Flour.

Figure 3 Proprietary formula A.

Figure 4 Proprietary formula B.

vitamins had been separately extracted from the sample by alkaline hydrolysis and methanol-Dichloromethane respectively. Standards of each vitamin to be analysed were prepared by accurately weighing 1 mg of the vitamin powder, dissolving it in 5mL deionised water and making it up to 10mL to form standard solutions of 0.1 mg/mL for each vitamin, however for standards of

CentralBringing Excellence in Open Access

Oche et al. (2017)Email:

J Hum Nutr Food Sci 5(3): 1116 (2017) 4/6

fat soluble vitamin, methanol was used as solvent . Each prepared sample and standard solution was stored by refrigerating in the dark at 4ºC and filtered through a 0.2 μm filter (Millex-GN) prior to injection into the HPLC column. The Chromatographic Conditions used were as shown below:

Parameter ConditionColumn for water-soluble vitamins: Acclaim PA, 3 μm, 3.0 × 150 mm

Mobile & stationary Phases: (i) 25 mM phosphate buffer (pH 3.6)(ii) CH3CN-mobile phase A (7:3, v/v)

Column for fat-soluble vitamins: Acclaim C18, 3 μm, 3.0 × 150 mm

Mobile and stationary phases:

(i) Methanol-methylcyanate (8 : 2 v/v)(ii) Methyl tert-butyl ether (MTBE)

Column Temperature: 25°C

Flow Rate: 0.5 mL/min

UV Detection wavelength: Absorption maximum of each vitamin

Injection Volume: 10 μL

Oxalate Determination: The permanganate titration method described by Ojinnaka [10] was used. A 2g portion of the sample was suspended in 100 mL distilled water and 5 mL 6 M HCl was added. The mixture was digested by heating at 100°C for 1 hour, then cooled and filtered. The mixture pH was adjusted to 4.5 by drop wise addition of concentrated aqueous ammonia solution before heating to 90°C in a water bath. This was then cooled and filtered (to remove ferrous ion precipitates). The filtrate was again heated to 90°C and 10 mL of 5% CaCl2 solution added with constant stirring and then allowed to cool and stored overnight in the refrigerator at 5°C. The mixture was then centrifuged at 3000×g for 5 min. The supernatant was decanted and the precipitate dissolved in 10 mL 20% H2SO4 . The solution was made up to 100 mL with distilled water and titrated against 0.05 M KMnO4 solution to a faint pink colour which persisted for

30 seconds. The oxalate content is given by the relationship that 1 mL of 0.05M KMnO4 solution corresponds to 0.00225 g oxalate [10]. The formula below was then used to calculate the oxalate content:

100 0.00225% titreOxalateW

× ×= (5)

Where, W represents the weight of sample used.

Phytate determination: The Oberlese spectrophotometer method described by Ojinnaka [10] was used. Two gram of the sample was extracted by acid hydrolysis, cold maceration and filtration with whatman filter paper to obtain the extract. Then 0.05 M standard phytate solution was prepared by weighing and dissolution. Also, 0.5 mL of the extract and 1 mL of standard phytate solution were put in separate test tubes, treated with 1 mL ferric ammonium sulphate solution, corked with stoppers and boiled in a water bath for 30 min. These were then cooled to 25 ºC in ice and 2 mL 2, 2-Bipyrimidine solution was added to each tube, mixed well and their respective absorbance measured with a UV-spectrophotometer at 519 nm. The phytate content of the sample was calculated by:

100%1000

Au C VtPhylateW As Va

× × ×=

× × × (6)

Where; W, Au, As, C, Vt and Va are weight of sample, absorbance of sample, absorbance of standard phytate solution, concentration of standardphytate (mg/mL), total extract volume and volume of extract used respectively.

RESULTS AND DISCUSSIONThe mineral composition of the cereal-legume composite

(CLC), proprietary infant formulae A, B, Formulated Infant food as well as the recommended daily allowance for each mineral are presented in Table 1. From the results, it is clear that the formulated food is highest in Na, Fe, Mg, Ca, K, Se and Zn compared with any of the foods analysed. This implies that more than any of the other foods analysed, the formulated food will better promote functions such as healthy red blood cell and bone development, tissue healing, fighting off diseases among other functions and this is in line with reports by Bio-organics Nutrient systems [13]. Also, it can be deduced that with exception of Cu, the formulated food can adequately provide the RDA of all the minerals analysed in infants fed as little as 150g per day. Pb and Ni were not detected. The implication is that they won’t be constituting toxins in infants fed with the formulated food.

Vitamins in the formulated food compared with nestle cerelac, friso-gold and

RDAs for infant: Table 2 shows the composition of water soluble vitamins in the Cereal-legume composite, Proprietary Formulae A, B and the Formulated infant food as well as the recommended daily allowance for each vitamin. This result shows that the formulated food is highest in vitamins B5, B6, B9, B12 and niacin compared with any of the foods analysed. The implication is that, more than any of the foods analysed, the formulated food can be expected to be most effective in promoting; healthy nervous systems, manufacture of red blood

Chicken Eggs

Hard Boil in Water for 20 mins and recover yolk

Cut yolk in pieces and oven-dry for 3 hours

Strain and oven-dry for 1 hour

Mill

Sieve with 80 µm mesh

Egg yolk Powder

Steep in excess fresh lime juice for 6 hours

Scheme 5 Preparation of Egg yolk Powder.

CentralBringing Excellence in Open Access

Oche et al. (2017)Email:

J Hum Nutr Food Sci 5(3): 1116 (2017) 5/6

Table 1: Mineral Composition of Cereal-Legume Composite, Proprietary Formulae A , B and Formulated Infant food

S/N MineralCereal-legume

Composite (mg/100g)

Proprietary Formula A (mg/100g)

Proprietary Formula B (mg/100g)

Formulated infant food (mg/100g)

RDA for infants (ages 6-59

months)1 Sodium 194 241 215 300.1 370 mg

2 Iron 5.12 8.45 6.24 8.96 10 mg

3 Magnesium 24.84 40.51 42.87 47.42 75 mg

4 Calcium 400.8 398 325 402.1 260 mg

5 Potassium 562 525 600.5 632 700 mg

6 Selenium 1.00* 3.00* 5.00* 9.00* 0.02 mg

7 Zinc 1.13 1.87 1.59 1.84 3.00 mg

8 Copper 0.02 0.02 0.01 0.02 0.22 mg

9 Lead N.D N.D N.D N.D N.A

10 Nickel N.D N.D N.D N.D N.A

* denote values are multiplied by 10-3

ND= Not detected as concentration was <0.0001 mg/100g

N.A= Not applicable

Table 2: Composition of Water-Soluble Vitamins in Cereal-Legume Composite, Nestle Cerelac, Friso- Gold and Formulated Infant food

S/N Water soluble Vitamins Cereal-legume composite (mg/kg)

Proprietary Formula A (mg/kg)

Proprietary Formula B (mg/kg)

Formulated infant food (mg/kg)

RDA for infants (6-59 Months)

1. Thiamin (B1) 1.34 3.89 0.34 1.15 0.3 mg

2. Ascorbic (C) 29.85 58.84 24.15 30.56 50.00 mg

3. Pyridoxine (B6) 0.28 0.34 0.31 0.39 0.30 mg

4. Nicotianamide (B3) 0.05 0.07 0.04 0.06 6.0 mg

5. Pantothenic Acid (B5) 0.03 0.05 0.04 0.06 1.8 mg

6. Folic acid (B9) 0.02 0.02 0.03 0.04 0.08 mg

7. Cyanocobalamin(B12) 3.00* 7.00* 6.00* 8.00* 5.00* mg

8. Riboflavin (B2) 1.16 3.84 0.68 2.14 0.50 mg

9. Niacin 2.07 3.71 2.57 3.93 4.00 mg

* denote values are multiplied by 10-4

Table 3: Composition of Fat-Soluble Vitamins in Cereal-Legume Composite, Nestle Cerelac, Friso- Gold and Formulated Infant food

S/N Vitamins Cereal-legume composite(mg/100g)

Proprietary Formula A(mg/100g)

Proprietary Formula B(mg/100g)

Formulated infant food(mg/100g)

RDA for infants(aged 6-59 Months)

1. Vitamin A, 0.23 0.33 0.26 0.34 0.40 mg2. Vitamin D, 3.00* 4.00* 7.00* 8.00* 5.00*mg3. Vitamin E, 2.96 3.89 3.56 4.03 5.00 mg4. Vitamin K, 0.03 0.04 0.03 0.04 3.00* mg* denote values are multiplied by 10-3

cells, energy release from protein, fat and carbohydrates in infants [13]. Table 2 also show that although Vitamins B2, B3 and C are highest in Proprietary Formulae A, they are higher in the formulated food than in the CLC and Proprietary Formulae B thus implying that the formulated food will better promote healthy tissue growth and development, immune system function and protein metabolism than the CLC and proprietary formula A [13].

It can be seen that with exception of Vitamins B5, B3 and B9, the formulated food will more than adequately provide the RDAs for water soluble vitamins in infants fed as little as 150g per day.

Composition of Fat-soluble Vitamins in the Formulated food Compared with Nestle Cerelac, Friso-Gold and RDA’s for infants: The Composition of fat-soluble vitamins in the cereals-legume composite, Proprietary Formula A, B and the Formulated

CentralBringing Excellence in Open Access

Oche et al. (2017)Email:

J Hum Nutr Food Sci 5(3): 1116 (2017) 6/6

Oche I, Patrice-Anthony O, Ubwa S, Adoga S, Inegedu A, et al. (2017) Micronutrients, Oxalate and Phytate Composition of a Potential Infant Food Made from Soybean, Yellow Maize, Short Rice, Egg Yolk and Crayfish. J Hum Nutr Food Sci 5(3): 11156.

Cite this article

infant food are as presented in Table 3. From the result in Table 3, it is clear that vitamins A, D and E are highest in the formulated food compared with any of the foods analysed. This implies that more than any of the foods analysed, the formulated food will better boost immune, cognitive and anti-oxidant functions, promote good vision, healthy cells, bones and teeth development in infants and this is in line with Bio-organics Nutrient systems report [13]. Also, it can be seen that Vitamin K in the formulated food (0.040 mg/100g) is higher than it is in the CLC and Proprietary Formula B but comparable with that in Proprietary Formula A (0.041 mg/100g). It is clear that the formulated food will sufficiently provide the RDA for fat-soluble vitamins in infants fed as little as 150g per day. This imply that the formulated infant food is an excellent source of micronutrient for infants who posses only a limited bowel since it is micronutrient-dense.

Phytate and oxalate content of the formulated food: The phytate and oxalate contents of the formulated infant food are as presented in Table 4. Both the phytate and oxalate levels are within the 0 to 5% safe permissible limits for phytates and oxalates in infant foods [7] and as such the chances of phytate and oxalate chelation of divalent mineral nutrients like Ca and Mg which are needed for bone development will be minimal.

CONCLUSIONThe study concludes that the potential infant food formulated

from soybean, yellow maize and short rice contains sufficient micronutrients for infants between ages 6-59 months as its vitamin and mineral compositions are in most instances, higher or comparable with that in the proprietary formulae studied and was found to be capable of providing infants with the RDAs for the micronutrients analysed except Cu. Also, its oxalate and phytate contents are below the maximum permissible limits for phytate and oxalates in food. It is recommended that this formulation be subjected to a feeding trial to really ascertain its field performance before recommending to mothers for use in infant feeding.

ACKNOWLEDGEMENTSThis research work was funded entirely by Tertiary education

trust fund (TETFUND).

REFERENCES1. World Health Organization. Children: Reducing Mortality.

2. Ahima KJ. Formulation of weaning foods using composite of maize, groundnut and soybean and assessing its nutritional effects using animal model (Unpublished master’s thesis). Kwame Nkrumah University of science and technology. Ghana. 2008; 1-37.

3. Solomon M. “Nutritive value of three potential complementary foods based on cereals and legumes”. A J Food Nutri Sci. 2005. 5: 1-7.

4. Almeida HD, Saldivar SO, Gomez MH, Rooney LW. “Production and Nutritional Value of Weaning Foods from Mixtures of Pearl Millet and Cowpeas”. Am Asso Cereal Chemists. 1993; 70: 14-18.

5. Sadana B, Chabra C. “Development and Sensory Evaluation of Low Cost Weaning Food Formulations”. J Hum Ecol. 2004; 16: 133-136.

6. Diallo M, Cisse Madi, Dessor F, Soulimani R, Oumar S, Desobry S. “Formulation of infant food based on local cereals: stability and effects on cognitive development”. J Nutri Food Sci. 2013; 211.

7. Guy E, Kyei N, Baffour Y, Ackah YP. “Nutrient content and sensory acceptability of a weaning diet formulated from mixtures of soya bean, groundnut and rice”. Afr J home economic edu. 2013; 1: 1-7.

8. Egli I, Davidsson L, Juillerat MA, Barclay D, Hurrell R. “The influence of soaking and germination on the phytase activity and phytic acid content of grains and seeds potentially useful for complementary feeding”. J Food Sci. 2002; 67: 3484-3488.

9. Anigo KM, Ameh DA, Ibrahim S, Danbauchi SS. “Nutrient composition of complementary food gruels formulated from malted cereals, soybeans and groundnut for use in North-western Nigeria”. Af J Food Sci. 2010; 4: 65-72.

10. Ojinnaka MC, Ebinyasi CS, Ihemeje A, Okorie SU. “Nutritional evaluation of complementary food gruels formulated from blends of soybean flour and ginger modified cocoyam starch”. Adv J Food Sci Tech. 2013; 5: 1325-1330.

11. Adebayo-Oyetoro AO, Olatidoye OP, Ogundipe FO, Ogundipe OO, Balogun OI, Bamidele FA, et al. “Evaluation of Proximate Composition and Functional Properties of Ofada Rice (Oryzasativa) Flour Blended with Bambara Groundnut (Vigna subterranean L)”. J Agri Vet Sci 3: 2011; 60-66.

12. Ikese CO, Okoye PAC, Ubwa ST, Akende S. “Proximate analysis and formulation of infant food from soybean and cereals obtained in Benue State, Nigeria”. Int J Food Sci & Biotec. 2017; 2: 106-113.

13. Bio-organics Nutrient systems. Enrich: what, why, how and where. BNS system publishers. Ibadan. 2005; 2-29.

Table 4: Phytate and Oxalate Contents of Formulated Infant Food

S/N Antinutrients Composition (%)MaximumPermissible-Limit (%)

1. Phytate 1.48 ± 0.05 0-5

2. Oxalate 0.78 ± 0.03 0-5