Comparative Evaluation of Nutritional Qualities of Nymphaea Lotus and Nymphaea Pubescens Seeds

International Journal of Biochemistry Research & Review

19(3): 1-10, 2017; Article no.IJBCRR.36536 ISSN: 2231-086X, NLM ID: 101654445

Comparative Evaluation of Nutritional Qualities of Nymphaea lotus and Nymphaea pubescens Seeds

M. Aliyu1, M. K. Atiku2, N. Abdullahi2, A. Zaharaddeen3 and A. A. Imam2*

1Department of Biochemistry and Molecular Biology, Federal University, Dutsin-Ma, Katsina, Nigeria. 2Department of Biochemistry, Bayero University, Kano, Nigeria. 3Department of Extension and Diagnostics, National Veterinary Research Institute, Vom, Plateau, Nigeria.

Authors’ contributions

This work was carried out in collaboration between all authors. Authors MA, AAI and MKA designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Author AZ managed the analyses of the study. Author NA managed the literature searches. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/IJBCRR/2017/36536 Editor(s): (1) Muhammad Abdul Rehman Rashid, Department of Plant Breeding and Genetics at “University of Agriculture Faisalabad”, Pakistan. Reviewers: (1) Celenk Molva, Izmir Institute of Technology, Turkey. (2) Patrícia Matos Scheuer, Federal Institute of Santa Catarina, Brazil. Complete Peer review History: http://www.sciencedomain.org/review-history/21426

Received 31st August 2017 th Original Research Article Accepted 4 October 2017 Published 16th October 2017

ABSTRACT

Aims: The study was aimed at evaluating the nutritional qualities of Nymphaea lotus and Nymphaea pubescens seeds. Study Design: It was designed to determine the proximate and amino acids profiles of Nymphaea lotus and Nymphaea pubescens seeds to indicate their nutritional potentials. Place and Duration of Study: Department of Biochemistry, Faculty of Basic Medical Sciences, Bayero University, Kano State-Nigeria, between August 2016 and January 2017. Methodology: Fresh seeds were collected, dried and grounded to smaller particle size. Standard official methods were employed in the proximate analysis while amino acids analysis was carried out using Technicon Sequential Multisample Amino Acid Analyser. Results: The result of the proximate composition revealed the richness of both seeds in carbohydrates, crude lipids and proteins. The amino acid analysis revealed that amino acids were more concentrated in Nympahaea lotus seed with the total value of 73.82 g/100 g than 70.70 g/100 ______

*Corresponding author: E-mail: [email protected]; E-mail: [email protected];

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g of Nymphaea pubescens seed. The former also exceeded the later in total essential amino acids by difference of 1.12 g/100 g. The highest scoring amino acid was arginine in Nymphaea lotus seed and leucine in Nymphaea pubescens seed while methionine was the most limiting amino acid in both the samples. All the essential amino acids in both samples satisfied WHO/ FAO reference protein except histidine, valine and methionine. The P-PER of Nymphaea lotus seed and Nymphaea pubescens seed were found to be 1.69 and 1.54 respectively, meaning Nymphaea pubescens seed may be slightly more bioavailable than the Nymphaea lotus seed. The Lys/Arg ratio of Nymphaea lotus seed (0.84) and Nymphaea pubescens seed (1.09) indicate their low arthrogenic potential. Conclusion: These results re-enforce the growing awareness that wild and semi-wild plant seeds can contribute useful amounts of essential nutrients to human diets.

Keywords: Proximate composition; amino acid; essential amino acid; protein.

1. INTRODUCTION Africa especially Nigeria [6]. Nymphaea lotus and Nymphaea pubescens fruits are either eaten raw Malnutrition is a condition that results from eating or cooked for food when dried by local people of a diet in which nutrients are either not enough or Northern Nigeria. This study evaluated and are too much such that the diet causes health compared the nutritional qualities of Nymphaea problem. It may involve calories, protein, lotus and Nymphaea pubescens seeds. carbohydrates, vitamins and minerals [1]. Not enough nutrients is called undernutrition or 2. METHODOLOGY undernourishment while too much nutrients is called overnutrition. Malnutrition is often 2.1 Collection and Identification of Plant specifically to refer to undernutrition where there Materials is not enough calories, protein and micronutrients [2]. There are number of causes of malnutrition. Nymphaea lotus and Nymphaea pubescens It results from: inadequate or unbalanced diet, seeds were obtained from Guzu- Guzu Dam in problems with digestion or absorption and certain Kabo Local Government Area, Kano state of medical conditions. Nigeria. The samples were authenticated by the

Nigeria has been ranked second after India in the Department of Plant Biology, Bayero University list of countries with highest cases of Kano with accession number BUKHAN 0356. malnourished children in the world as it accounts for 10% of the 160 million stunted children 2.2 Preparation of Samples globally. The malnutrition in Nigeria is not very encouraging as about 37% of the children are The samples were thoroughly washed to remove stunted, children under the age of five have sand and the drained parts were air dried later chronic malnutrition and 29% are underweight, using drier (Drier, AD61, UK). The samples were while 18% have acute malnutrition [3]. However, grounded using wooden mortar and pestle until protein energy malnutrition affects children the powder was obtained to ensure homogeneity. most because they have less protein intake. The The fine powdered samples were then stored at few rare case found in the world are almost 25°C into labeled plastic containers prior to use. entirely found in children as a result of diets or ignorance in the nutritional needs of children 2.3 Proximate Analysis particularly in the cases of milk [4]. 2.3.1 Determination of moisture content [7] Nymphaea lotus (white water lily) and Nymphaea pubescens (red waterlily) also known as Bado Three clean dried petri dishes were weighed gero and Bado dawa in the native Hausa (W1) using weighing balance (A & D weighing, language, are perennial plant that grows up to 45 HR250AZ, USA) and 5 g of the pulverized cm in height. They are herbaceous aquatic sample was placed in each of them and weighed plants, whose leaves float or are submerged in (W2). They were placed in an oven (Tricity water [5]. These plants prefer clear, warm and Bendex, S1530RD, UK) at 100°C for 17 hours. slightly acidic water and is localized to central The dishes were removed and cooled in a and Southern Europe, Asia, the Middle East, dessicator for 30 minutes and finally weighed Northern Africa, and Tropical Mountain of West (W3).

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Calculation hexane into the weighed extraction flask. The extraction was then carried out for 3 hours using The loss in weight due to drying is equal to the the heating mantle at 60-80°C making sure there moisture content of the sample, was continuous flow of water in the condenser (which served as the cooling system). The sample was then removed, air-dried and then %moisture = × 100 placed in an oven (Tricity Bendex, S1530RD, UK) at 80°C until a constant weight was obtained W W  2 3 100 (W3). W W 2 1 Calculation

Where (─) Crude fat (%) = × 100 W1 = weight of empty petri dish W = weight of empty petri dish + sample 2 Where: W = weight of empty petri dish + sample after 3 drying W1 = Weight of sample W = Weight of sample and filter paper 2.3.2 Determination of ash content [8] 2 W3 = Weight of sample after lipid extraction and filter paper Porcelain crucible or silica disc was ignited in a hot Bunsen burner flame or in a muffle furnace 2.3.4 Determination of crude protein [7] for about one minute; it was then transferred into a dessicator for cooling and weighing (W1). The Exactly 0.15 g of dried (moisture free) sample sample (2 g) was placed into the crucible and was weighed and transferred into the Kjeldahl weighed again (W2). A crucible containing the digestion flask (30-35 ml). 0.8 g of catalyst (0.7 g sample was heated gently on a muffle furnace at sodium sulphate, 0.06 g copper sulphate and 550-570°C to burn off all the organic matter. The 0.04 g mercury (II) oxide red) was added into the carbon char burn off as C02 leaving a white ash. digestion flask. To this mixture was added 2 ml of The crucible taken out was immediately covered concentrated sulphuric acid of which color turned and placed in a dessicator for cooling and then black. The mixture in the digestion flask was weighed (W3). heated on the heating mantle for 1 hour until liquid became clear green. The digest was Calculation cooled and made alkaline with 15 ml 40% NaOH. The digest was then transferred to the steamed W W out apparatus using minimum volume of water. Ash %  3 1 100 W W The ammonia steamed is distilled into 10 ml 2% 2 1 boric acid solution with 5 drops of methyl red indicator for 15 minutes. The distilled ammonia Where was titrated with 0.02M hydrochloric acid. The same procedure was carried out for the blank. W1 = weight of crucible W2 = weight of crucible + sample Calculation W3 = weight of crucible after all organic matter was burnt off . × × %Nitrogen = 2.3.3 Determination of crude fat using soxhlet extraction method [7] Where: V= volume of acid used (titre value) Exactly 3 g of the sample was carefully weighed W= weight of sample= 0.15 g

(W1) into a folded fat-free filter paper using %Crude protein= %Nitrogen × 6.25 weighing balance (A & D weighing, HR250AZ, (multiplication factor) USA). This was properly tied with a thread at both ends and weighed (W2). This was carefully 2.3.5 Determination of crude fiber content [7] placed in the extraction thimble and small cotton wool placed on top. The whole apparatus was The sample (1.00 g) was weighed into a flat- 3 then connected after the addition of 300 ml of bottom flask, 100 cm of 20% H2SO4 was added

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to the sample and was boiled on hot plate for 30 the ration of 10:5:1 was added into the flask to minute. The content was filtered with filter paper facilitate digestion. Four pieces of anti-bumping and the residue was boiled again in 100 cm3 2M granules were added. The flask was then placed NaOH solution for 30 minutes and filtered. The in Kjeldhal digestion apparatus for 3 hours until residue was then dried, weighed and ashed. The the liquid turned light green. The digested crude fibre content was then calculated using the sample was cooled and diluted with distilled formula: water to 100 ml in standard volumetric flask. Aliquot (10 ml) of the diluted solution with 10 ml Crude fiber (%) = × 100 of 45% sodium hydroxide was placed into the Markham distillation apparatus and distilled with 10 ml of 2% boric acid containing 4 drops of Where: methyl red indicator until 70 ml of distillate was

collected. The distillate was then titrated with W = Weight of sample extracted filter paper 1 standardize 0.01N hydrochloric acid to grey W2 = Weight of “W” after ashing. coloured end point, the percentage nitrogen in

2.3.6 Estimation of total carbohydrates the original sample was calculated using the formula:

Total carbohydrate was estimated by calculation/ ()×.×× difference [8]. The available carbohydrate of % N = × 100 samples was estimated by ‘difference’. In this, × the sum of the percentages of all the other proximate components was subtracted from 100. Where: i.e. a = Titre value of the digested sample

b = Titre value of blank sample % Carbohydrates = 100- (% Crude Fat + % v = Volume after dilution (100 ml) Crude Protein + % Ash + % Crude fibre). w = Weight of dried sample (g) 2.4 Analysis of Amino Acid Profile of c = Aliquot of the sample used (10 ml) Nymphaea lotus and Nymphaea 14 = Nitrogen constant in g

pubescens Seeds 2.4.3 Hydrolysis of the samples

The amino acid profile of the Nymphaea lotus and Nymphaea pubescens seeds were The defatted sample (1.204 g) was weighed into determined as previously described by [9]. The glass ampoule. 7 ml of 6N HCl was added and samples were dried to constant weight, defatted, oxygen was expelled by passing nitrogen into the hydrolysed, evaporated in a rotary evaporator ampoule (This is to avoid possible oxidation of and then loaded into the Technicon Sequential some amino acids during hydrolysis). The glass Multisample Amino Acid Analyser (TSM). ampoule was then sealed with Bunsen burner (Technicon Instruments Corporation, New York). flame and placed in an oven preset at 105°C ± 5°C for 22 hours to effect hydrolysis. The 2.4.1 Defatting of sample ampoule was allowed to cool before it was broken open at the tip and the content was A known weight of the dried sample was weighed filtered to remove the humins. The filtrate was into extraction thimble and the fat was extracted then evaporated to dryness at 40°C under with chloroform/methanol mixture in ratio of 2: 1 vacuum in a rotary evaporator. The residue was using soxhlet extraction apparatus as previously dissolved with 5 ml of acetate buffer (pH 2.0) and described by [7]. The extraction lasted for 15 stored in plastic specimen bottles, which were hours. kept in deep freezer.

2.4.2 Percentage nitrogen determination 2.4.4 Loading of the hydrolysate into the TSM analyzer The sample (2 g) was weighed, wrapped in whatman filter paper (No.1) and put in the The amount loaded was 60 microlitres. This was Kjeldhal digestion flask. Concentrated sulphuric injected into the cartridge of the analyzer. The acid (10 ml) was added. Catalyst mixture (0.5) TSM analyzer is designed to analyse free acidic, containing sodium sulphate (Na2SO4), cupper neutral and basic amino acids of the hydrolysate. sulphate (CuSO4) and selenium oxide (SeO2) in The period of an analysis lasted for 75 min. The

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amino acids present in the samples were The tryptophan in the known sample was identified by matching their peak retention time in hydrolyzed with 4.2 M Sodium hydroxide. The the chromatogram with those peaks of standard known sample was dried to constant weight, mixture of amino acids with norleucine as internal defatted, hydrolyzed, evaporated in a rotary standard. evaporator and loaded into the Applied Biosystems PTH Amino Acid Analyzer as 2.4.5 Calculating the amino acid values from described above. the chromatogram peaks 2.4.7 Estimation of seed samples protein An integrator attached to the analyzer calculates quality the peak area proportional to the concentration of each of the amino acids. The net heights of each To estimate the quality of dietary protein in the peak produced by the chart recorder of the TSM seed samples, the Total Essential Amino Acid (each representing an amino acid) were (TEAA), Total Non-Essential Amino Acid measured. The half-height of the peak on the (TNEAA), Total Sulphur Amino Acid (TSAA), chart was found and the width of the peak at the percentage of cysteine in TSAA (%Cys/TSAA), half- height was accurately measured and result Total Aromatic Amino Acid (TArAA), Leu/Ile ratio, recorded. Approximate area of each peak was Lys/Arg ratio etc, were calculated as described then obtained by multiplying the height with the by FAO [10]. The Predicted Protein Efficiency width at half height. The norleucine equivalent Ratio (P-PER) was also determined using the (NE) for each amino acid in the standard mixture equation of Alsmeyer’s. I.e. P-PER = - 0.468 + was calculated using the formular: 0.454 (Leu) – 0.105 (Tyr) [11]. The percentage amino acid scores of the seed samples were also ( ) NE= calculated using the formula: ( ) % Amino acid score = [Amount of amino acid per Constant S was calculated for each amino acid in test protein (g/100 g)/ Amount of amino acid per the standard mixture: protein in FAO/WHO (1995) reference pattern (g/100 g)] × 100 Sstd. = NEstd. × Mol. weight × µMol.AAstd. 3. RESULTS AND DISCUSSION Finally the amount of each amino acid present in the samples was calculated in grams per 100 g 3.1 Proximate Composition protein using the following formula: Table 1 presents the proximate composition of Concentration (g/100 of protein) = NH × NH/2 × the Nymphaea lotus and Nymphaea pubescens Sstd. × C seeds. The moisture, crude fibre contents of Nymphaea lotus and Nymphaea pubescens Where: seeds are not significantly different (p>0.05). However, ash, lipid, crude proteins and [ ×] carbohydrates contents of the Nymphaea lotus C = ()× . ×() seed and Nymphaea pubescens seeds recorded significant difference (p<0.05). Where Wt= weight, NH = net height, W = width, nleu = norleucine The percentage of ash, moisture, crude protein, crude fat (lipids), crude fibre and carbohydrate 2.4.6 Determination of tryptophan contents of Nymphaea lotus and Nymphaea pubescens seeds are presented in Table 1. The Tryptophan is a difficult amino acid to determine Nymphaea pubescens seed had higher in proteins and peptides because it chemically percentage value of ash content compared to decomposes during acid hydrolysis. It should be Nymphaealotus seed. This is higher than the noted that tryptophan is destroyed by 6N HCL reported value of Nymphaea lotus seed by [12,5]. during hydrolysis. Alkaline hydrolysis was Thus, the result indicated that Nymphaea lotus improved by using sodium hydroxide (NaOH) and Nymphaea pubescens seed could serve as instead of barium hydroxide to prevent problems good source of mineral element since ash with both precipitation and adsorption of content measured the minerals content of food tryptophan. substances and were found to be within the

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range reported for some leguminous seed [13]. range reported by [16,5,12] for Nymphaea lotus However, the percentage for the moisture seeds. Thus, ingestion of foodstuff which has content in Nymphaea lotus and Nymphaea reasonable amount of dietary fiber reduce pubescens seeds were 5.31% and 6.20% postprandial glucose response after respectively which is less than 8.5% in Annona carbohydrate rich meals as well as lowering total muricata and 9.7% in pride of Barbados seeds LDL cholesterol levels [15]. reported by [13,14] respectively. These results are comparable to the literature value of 6.00% Similarly, the carbohydrate content of both reported for Nymphaea lotus seed [12]. However, Nymphaea lotus and Nymphaea pubescens the results were lower than 8.20% reported by seeds were lower compared to the reported [5]. The low moisture content of both Nymphaea value in Nymphaea lotus seed (80.96%) by [12]. seeds may signify longer shelf life since seeds The difference in carbohydrates contents could with moisture content greater than 15% are be attributed to difference in climate and soil subjected to deterioration from mould growth, condition. The carbohydrate composition present heat, insect damage and sprouting [13]. in both Nymphaea seeds is enough to prevent Moreover, the percentage of crude fat (lipids) in ketosis as breakdown of 1 gram of (glucose) Nymphaea lotus seed was found to be higher carbohydrate yield 4 kcal of energy [17]. Similar than in Nymphaea pubescens seeds. In addition carbohydrate content was reported by [12] had earlier reported that Nymphaea lotus Mohammed et al. [16] which reported that, seeds seeds have a lipid or crude fat content of 9.33% of Nymphaea lotus serves as good source of which was lower than value reported in the carbohydrate and is enough to prevent ketosis. present study. Therefore, both Nymphaea seeds The role of Carbohydrate is not limited to energy could serve as the secondary source of energy. intake as they also play a vital role in both the taste of the food and the pleasure of eating Table 1. Proximate compositions of [18]. Nymphaea lotus seed and Nymphaea pubescens seeds Therefore, the proximate composition of the Nymphaea lotus and Nymphaea pubescens Parameters Nymphaea Nymphaea seeds indicates they are highly nutritious as they (%) lotus seed pubescens contains high protein content hence could seed supplement other protein sources such as beans, Moisture 5.31±0.32 6.20±0.87 peas and groundnuts especially in dry seasons a a Ash 1.33±0.11 3.00±0.30 and in arid regions [19] when or where most a a Proteins 4.92±0.34 4.14±0.06 common protein sources might be scarce and a a Lipids 13.23±1.01 9.28±0.15 expensive, hence could help to reduce Fibre 5.17±0.51 5.00±0.26 malnutrition and food insecurity a a Carbohydates 75.35±1.38 78.58±0.89 All values are expressed as mean ± standard 3.2 Amino Acids Compositions deviation for three determinations. Rows with similar superscript are statistically significant (p<0.05) The amino acid composition and percentage Furthermore, Nymphaea lotus seed was found to Amino acid score of Nymphaea lotus and have higher crude protein content than the Nymphaea pubescens seeds are presented in reported value of (3.09%) by [5]. It was earlier Table 2. The result indicates the presence of all reported that Nymphaea lotusseed could be the essential amino acids while two non-essential good source of protein [15]. The percentage amino acids were totally absence in the two value of crude protein obtained for Nymphaea samples screened. pubescens seed was slightly lower than in Nymphaea lotus seed. Thus, this study suggests From the various parameters presented in Table that Nymphaea lotus seeds possess high protein 2, total amino acids (TAA), total non-essential content compared to Nymphaea pubescens amino acids (TNEAA), total essential amino seeds. Proteins ensure proper growth and also acids (TEAA), their respective percentages, the serve as source of some essential amino acids predicted protein efficiency ratio (P-PER), Leu/Ile for body use. Analysis revealed slightly higher ratio and Essential amino acid index (EAAI) percentage of crude fibre in Nymphaea lotus values of Nymphaea lotus and Nymphaea seed than obtained in the Nymphaea pubescens pubescens seed were calculated and presented seeds. However these values are within the in Table 3.

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The amino acids contents of Nymphaea lotus All essential amino acids in both Nymphaea lotus seed and Nymphaea pubescens seed were and Nymphaea pubescens seeds were found to presented in Table 2 showing only eighteen satisfied the FAO/WHO ideal protein value in amino acids out of the common twenty amino children except for valine, histidine and acids found in proteins, including ten essential methionine which were closer to the value amino acids and eight non-essential amino acids. established by FOA/WHO ideal protein in This may be due to the conversion of the amide children. Hence, with proper processing, the glutamine and asparagine to their corresponding seeds could meet up with the WHO ideal protein amino acids, glutamate and aspartate value for both children and adults. respectively [20].

Table 2. Amino acid composition and percentage amino acid score of Nymphaea lotus and Nymphaea pubescens seeds

Amino acid Concentration (g/100 g) FAO/WHO Ref. %Amino acid score N. lotus N. pubescens Standard N. lotus N. pubescens Leucine 5.49 5.14 4.2 130.71 122.38 Lysine 4.48 4.88 4.2 106.67 116.19 Isoleucine 4.52 4.19 4.2 107.62 99.76 Phenylalanine 2.48 2.39 2.3 107.83 103.91 Tryptophan 1.21 0.92 1.1 110.00 83.63 Valine 4.01 3.71 4.2 95.48 88.33 Methionine 1.23 1.20 2.2 55.91 54.55 Arginine 5.33 4.47 4.0 133.25 111.75 Histidine 2.43 2.20 3.4 71.47 64.71 Threonine 3.16 3.25 2.8 112.86 116.71 Cysteine 1.15 1.27 2.0 57.50 63.50 Proline 3.55 3.96 - - - Tyrosine 3.10 3.10 2.8 110.71 110.71 Alanine 4.02 3.41 - - - Glutamic acid 12.11 11.58 6.3 192.22 183.81 Glycine 3.61 3.94 - - - Serine 3.94 3.59 - - - Aspartic acid 8.00 7.50 - - - - Means not reported/Not determined

Table 3. Nutritional quality indices of protein

Amino acid grouping Nymphaea lotus seed Nymphaea pubescens seed Total amino acid (TAA) 73.82 70.70 Total non-essential amino acid (TNEAA) 39.48 38.38 Total essential amino acid (TEAA) -with His 34.34 32.32 -without His 31.91 30.12 % TNEAA 53.48 54.29 % TEAA -with His 46.52 45.71 -without His 43.23 42.60 Total Sulphur amino acid (TSAA) 2.38 2.47 % TSAA 3.22 3.49 % Cys in TSAA 48.32 51.42 Total Aromatic amino acid (TArAA) 6.79 6.41 1.69 1.54 9.07 Leu/Ile ratio 1.21 1.23 Lys/Arg 0.84 1.09 Essential amino acid index(EAAI) 87.6 84.2

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Among the essential amino acids, leucine had g/100 g). This difference could be attributed to the highest concentration in both the samples, genetic differences among the Nymphaea followed by isoleucine. The least essential amino species and the values obtained in the present acid is methionine with Nymphaea lotus seed study are comparable with that of some plant having the higher content. Nymphaea lotus seed foods which range between 39.3 – 76.5 g/100 g had higher concentration of all amino acids protein as reported by earlier workers [25,26]. compared to Nymphaea pubescens seed except for lysine, cysteine, proline and glycine. This The predicted protein efficiency ratio (P-PER) could be attributed to genetic difference between value is one of the quality parameters used in the the samples. evaluation of proteins [21]. The P-PER of Nymphaea lotus seed and Nymphaea pubescens The arginine content of Nymphaea lotus and seed were found to be 1.69 and 1.54 Nymphaea pubescens seeds was higher than respectively, meaning Nymphaea pubescens the FAO/WHO [21] recommendations for infants seed may be slightly bioavailable than the (Arginine 4.0 g/100 g). Arginine is good for Nymphaea lotus seed. This is slightly lower than children and it is considerably high in both the the reported values for true digestible protein of Nymphaea lotus and Nymphaea pubescens whole dried honey bees (Apis mellifera L.), 2.47 seeds with respectively. Isoleucine is an and 2.50 for casein [27]. Due to the fact that the essential amino acid both for young and old. experimentally determined PER usually ranged Methionine is needed for the synthesis of choline from 0.0 for a very poor protein to a maximum which in turn forms lecithin and other possible value of just over 4 [28], the result of phospholipids in the body. When the diet is low in this work shows that both Nymphaea lotus and protein, for instance in alcoholism and Nymphaea pubescens seeds could be efficiently kwashiorkor, insufficient choline may be formed; utilized in human body. this may cause accumulation of fat in the liver [22]. Furthermore, the Leu/Ile ratio value of Nymphaea lotus and Nymphaea pubescens seeds were The amino acid scores(a rating of the quality of a found to be 1.21 and 1.23 respectively, of which test protein by comparing its amino acid pattern Ile was less than half that of Leu. It has been with that of FAO/WHO reference ideal protein for suggested that an amino acid imbalance from pre-school children) of Nymphaea lotus and excess leucine might be a factor in the Nymphaea pubescens seeds are shown on development of pellagra due to sorghum Table 2. The highest scoring essential amino consumption [29]. High Leucine in the diet acid was Arginine (133.25%) in Nymphaea lotus impairs tryptophan and niacin metabolism and is seed and leucine in Nymphaea pubescens seed responsible for niacin deficiency in sorghum while methionine was the lowest scoring amino eaters [30]. acid (55.91 and 54.55%) in both the samples making it the first limiting amino acid. This is in The Lysine/Arginine (Lys/Arg) ratio has been conformity with the report that the essential reported to play a role in the artherogenocity of a amino acids most often acting in a limiting protein. [31] Reported that a Lys/Arg ratio as high capacity are Methionine and Cysteine, Lysine as 2.0, increases the artherogenic potential of a and Tryptophan [23]. Leucine, Isoleucine and diet. The Lys/Arg ratio of the Nymphaea lotus Tryptophan are also present in significant (0.84) and Nymphaea pubescens seed (1.09) are quantities. Methionine is needed for the much lower than that of casein with Lys/Arg ratio synthesis of other important substances including 2.0. A similar observation has been made for choline. The intake of methionine may prevent Kerstingiella geocarpa [32]. Thus, since the fatty liver and causes development and Nymphaea lotus and Nymphaea pubescens growth [22]. seeds showed a low Lys/Arg ratio, the samples will possess a low artherogenic potential. Furthermore, Table 3 shows the nutritional quality determinations of Nymphaea lotus and The essential amino acid index (EAAI) and Nymphaea pubescens seeds. The nutritive value nutritional index of Nymphaea lotus seed were of a protein depends primarily on the capacity to higher than the Nymphaea pubescens seed. satisfy the needs for nitrogen and essential However, the nutritional indexes of the both amino acids [24]. The total amino acids, TAA Nymphaea seeds were much higher compared to (73.82 g/100 g) of Nymphaea lotus seed is the A. bisporus (13.69%) and P. florida (12.59%) higher than Nymphaea pubescens seed (70.70 as reported by [33]. The essential amino acid

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index can be useful as a rapid tool to evaluate and nutritional ignorance. Archives of food formulations for protein quality [34]. Dermatology. 2012;1375:630-636. However, it does not account for differences in 5. Musa A, Birnin-Yauri A, Muhammad C, protein quality due to various processing Umar A. Proximate composition and methods or certain chemical reactions [34]. mineral analysis of Nymphaea lotus seeds. Protein material is said to be of good nutritional African Journal of Food Science and quality when its essential amino acid index Technology. 2012;3(7):4. (EAAI) is above 90% and to be useful as food 6. Akinjogunla OJ, Yah CS, Eghafona NO, when the values are around 80% and are Ogbemudia FO. Antibacterial activity of inadequate for food material when values are leave extracts of Nymphaea lotus on below 70% [35]. From the present study it is Methicilin Resistant Staphylococcus observed that the EAAI values were generally aureus (MRSA) and Vancomycin-Resistant appreciable and could be attributed to the fact Staphylococcus aureus (VRSA), isolated that Nymphaea seeds, a wild haebaceous plant, from clinical samples. Annals of Biological possessed most essential amino acids in Research. 2010;2:174-184. appropriate quantities. 7. AOAC. Official methods of analysis. Association of Official Analytical Chemists, 4. CONCLUSION Washington, D.C., USA. 1997;17:807-928. 8. AOAC International. Official method of From the results obtained from this work, it can analysis of AOAC. 17th Edition current be concluded that Nymphaea lotus and through 1st revision Gaithersburg, MD, Nymphaea pubescens seeds are of high USA, Association of Analytical nutritional qualities. Both the seeds are rich in Communities; 2002. essential amino acids that if consumed in 9. Benitez LV. Amino acid and fatty acid sufficient amount, it could contribute to meeting profiles in aquaculture nutrition studies, P. human nutritional needs and helps to combat 23-35. In S.S. De Silva (ed.) Fish Nutrition diseases associated with malnutrition. The level Research in Asia. Proceedings of the Third of arginine and histidine in the seeds warrants Asian Fish Nutrition Network Meeting. them to be recommended for children food since Asian Fish. Society Special Publication. children need arginine and histidine in their 1989;4:166. foods. This nutritional information could be of 10. AOAC. Official Method of Analysis of the great use to nutritionists, industrialists, Association of Official Analytical Chemists, researchers, policy makers development Washington, D. C., AOAC. Asian Fisheries agencies and encourage the consumption of Society, Manila Philippines. 18th Edition. Nymphaea lotus and Nymphaea pubescens 2010;810-931. seeds so that they become part of normal diet 11. Alsmeyer RH, Cunningham AE, Happich rather than being considered as ‘famine’ or ‘poor ML. Equations to predict PER for amino peoples’ food especially in arid regions. acid analysis. Food Technol. 1974;28;34-

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