Minerals, Vitamins and Amino Acids Compositions of Melanthera Biflora

International Journal of Biochemistry Research & Review

24(4): 1-7, 2018; Article no.IJBCRR.34900 ISSN: 2231-086X, NLM ID: 101654445

Minerals, Vitamins and Amino Acids Compositions of Melanthera biflora

E. C. Kalu1*, O. C. Ikewuchi1 and C. J. Ayalogu1

1Department of Biochemistry, Faculty of Science, University of Port Harcourt, P.M.B. 5323, Choba, Rivers State, Nigeria.

Authors’ contributions

This work was carried out in collaboration between all authors. All authors read and approved the final manuscript.

Article Information

DOI: 10.9734/IJBCRR/2018/v24i430062 Editor(s): (1) Dr. Fatih Oz, Professor, Department of Food Engineering, Agriculture Faculty, Ataturk University, Turkey. Reviewers: (1) Anna Leśniewicz, Analytical Chemistry and Chemical Metallurgy Division, Faculty of Chemistry, Wrocław University of Science and Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland. (2) Irfan Ur Rauf Tak, Centre of Research for Development, University of Kashmir, India. (3) M. H. Shagal, Department of Chemistry Modibbo Adama University of Technology Yola, Adamawa State, Nigeria. Complete Peer review History: http://www.sdiarticle3.com/review-history/34900

Received 03 November 2017 Accepted 24 January 2018 Original Research Article Published 21 February 2019

ABSTRACT

The minerals, vitamins and Amino acids composition of Melanthera biflora leaves were investigated using standard methods. The result shows that out of the eleven vitamins detected; only vitamin C was found in moderately high amount (99.37 mg/100 g), while the rest were in insignificant amount. Ten minerals element were detected, including iron (14.63) copper (2.32). Cobalt (0.04) manganese (5.13), calcium (93.52) magnesium (126.17), potassium (71.39) sodium (32.37), phosphorus 36.50, selenium (0.006) all in mg/100 g. The leaves have essentials and non-essential amino acid of 47.5% and 45.94% respectively. It has all the essential amino acids except tryptophan. This result shows that the leaves contain an appreciable amount of amino acids, vitamins and minerals that could be included in diets to supplement our daily dietary allowance.

Keywords: Minerals; vitamins; amino acids; essentials.

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

Kalu et al.; IJBCRR, 24(4): 1-7, 2018; Article no.IJBCRR.34900

1. INTRODUCTION were grounded into a fine power using a mortar and a pestle and sieved to pass through a 40- Green leafy vegetables occupy an important mesh sieve and stored in an air-tight container place among the food crops as they provide under refrigerated temperature for further use. adequate amount of vital nutrients, vitamins and minerals for human consumption. They are also 2.2 Determination of Mineral Composition rich source of carotene, ascorbic riboflavin, folic acids, among others. They also contain many 2.2.1 Ashing minerals like calcium, iron and phosphorus [1]. The pulverized sample (1.0 g) was weighed into Plants constitute an important source of natural a petri dish. The sample in the petri dish was products which differ widely in terms of structure emptied into the muffle furnace pot. The pot and and biological properties. They have played a its contents were placed in the furnace at 550°C remarkable role in food industries, local for 4 hours. The sample in the pot was allowed to consumption as well as in the orthodox medicine cool in the dessicator. The process was repeated of various countries. until a constant weight was attained.

Generally vegetables species abound in the 2.3 Determination of Mineral Composition world. Green leafy vegetables constitute an indispensable constituent of human diet in Africa and in West Africa in particular [2]. Nigeria is 2.3.1 Ashing blessed with great natural tropical rain forest that is characterized with viable soil where vegetables The pulverized sample (1.0 g) was weighed and of high nutritional value are grown. Most of these place in the petri dish. The sample in the petri vegetables are not well known, hence not dish was emptied into the muffle furnace pot. The consumed as their nutritional compositions are pot and its content were placed in the furnace at not known. 550”C for 4 hours. The sample in the pot was allowed to cool in the desiccators. The process Melanthera biflora, is a perennial herbaceous was repeated until a constant weight was plant which belong to the family of asterecae. Its obtained. common name is beach daisy. It is known among the Igbos in south-eastern Nigeria as “akwuwa” 2.4 Mineral Analysis and “akwuba” among the Efiks in Cross Rivers State of Nigeria. It produces luxurious, edible The ashed sample (0.2 g) was weighed into the leaves which are used in making soup. pre-cleaned borosilicate 250 ml capacity beaker for digestion. 30 ml of the nitric acid was added The present study therefore is aimed at into the weighed sample in the beaker. The evaluating the level of mineral elements, vitamins sample within the digesting solvent was placed and amino acids composition of Melanthera on the hot plate for digestion in the fume biflora, a tropical leafy vegetable found in south cupboard. The beaker and its content after the eastern Nigeria so as to encourage its digestion were allowed to cool. Another 20 ml of consumption in other to enhance healthy living. the digesting solvent was added further in the fume cupboard and the mixture were allowed to 2. MATERIALS AND METHODS cool to the room temperature. The mixture was filtered into the 250 ml volumetric capacity 2.1 Sample Collection and Preparation borosilicate container. The filtrate was made up to the mark with the deionized water. The same The leaves of Melenthera biflora were harvested procedure was followed for the digestion of the fresh from “Ude” plantation in okon, Aku, in other samples. Ohafia Local Government of Abia State and was later identified by a taxonomist in the herbarium All the digested samples were sub-sampled into of the department of plant science, University of pre- cleaned borosilicate glass containers for Port Harcourt, Dr. Edwin Nwosu. atomic Absorption Spectrophotometer analysis.

The harvested vegetable leaves were destalked, Standards of iron, copper, cobalt, manganese, washed with cold running tap dried in an oven at selenium, calcium, magnesium, potassium and 600C for 24 hours, after the drying the leaves sodium solutions of 0.2,0.4,0.6,0.8 and 10 mg/ml

Kalu et al.; IJBCRR, 24(4): 1-7, 2018; Article no.IJBCRR.34900

were made from the each of the heavy metals 2.7 Determination of Amino Acid solution of 1000mg/c stock solutions of be analytes. The set of standard solutions and the Extraction: Modified AOAC method 982.30 2006 filtrate of the digested samples were analyzed was followed in the extraction of the sample for by AAS. The detection limit of the metals the amino acid analysis. in the sample was 0.000/mg/l by means of the UniCAM 929 London, Atomic Absorption The dried and pulverized sample was made to be Spectrophotometer powered by the SOLAR free of water by ensuring constant weight for a software. Iron, copper, cobalt, manganese, period of time in the laboratory. selenium, calcium, magnesium, Potassium and sodium cathode lamps were used for the The sample of 10.0g was weighed into the 250ml analysis of the respective minerals ions in the conical flask capacity. The sample was defatted standard and the filtrate of the samples. Gas by extracting the fat content of the sample with mixtures were used in the generation of the 30ml of the petroleum spirit three times with flame. soxhlet that was equipped with thimble. The sample was hydrolyzed three times for complete hydrolysis to be achieved. 2.5 Determination of Phosphorus The amino acid content of the sample was The sample (25 mg) was weighed into the recovered by extracting with 30 ml of the schoniger flask and burnt in excess of oxygen dichloromethane three times before gas. The product was digested with nitric acid. concentrating to 1.0 ml. The concentrated extract The content was boiled for a minute to ensure was derivastized for volatility that is suitable for complete conversion of phosphorus pentoxide to gas chromatography. The GC conditions for orthophosphate. amino acids is as follows.

The solution was passed through a 10 cm long Injection temperature : split injection, Split Ratio: resin column, and the filtrate was collected, in a 20:1Carrier Gas: Hydrogen Flow Rate: 10 ml Pyrex test tube, 2 ml of the colour 1.0ml/min, Inlet Temperature: 2500C Colum development reagent was added for the Type: HP5Column Dimensions: 30m x 0.25 mm absorbance reading at 650 mm for both the x 25m Over program: initial at 600C, first Ramp at standards and the sample filtrate. 80C/min for 20 min, maintained for 2min second Ramp at 120C/min 6 min, constant for 2 mins, 2.6 Vitamin Determination of Vitamin Detector: FFPD, Detector Temperature: 3200C, Composition Hydrogen Pressure 20 Psi, Compressed Air:35 Psi. 2.6.1 Sample preparation 3. RESULTS AND DISCUSSION The Vitamins content of the sample was analyzed by a modification of AOAC Methods The leaves of Melanthera biflora had higher 992.03, 992.04 and 999.26 [3]. The sample was calcium, sodium and magnesium (Table 1) than made to attain the laboratory atmospheric A. hybridus as reported by Oguntana [4], but conditions on the bench after removing the lower potassium content. From the above result, samples from the storage chamber at less than we can infer that Melanthera biflora are sources or equal to 4◦C.The sample was pressed in the of soluble minerals, which help in the mortar carefully to avoid forming balls. The maintenance of acid-base balance of the body homogenized sample (0.10g) was weighed into a tissues. These minerals help complete the beaker. After the extraction, the extracts were absorption of vitamins, proteins, fats and concentrated to 1.0 ml for the chromatographic carbohydrates of the food [5]. Calcium and iron analysis. GC condition of analysis were as are important elements needed by the cells and follows: Injection temperature: split injection, Split tissue of the bodies, especially as they form Ratio 20:1, Carrier gas: Nitrogen, Flow Rate: 1.0 component of enzymes and electron transducers ml/min, Inlet Temperature: 2500C, Column Type: [6]. Calcium is required for bone and teeth HP5, Column Dimensions: 30m x 0.25 mm x formation and in the proper functioning of the 0.25 mm, Over program: Initial Temperature at nervous system. The high calcium concentration 500C, For 2 minutes, Detector: PFPD, Detector in Melanthera biflora suggests that it could be Temperature: 3200C Hydrogen Pressure: 20Psi, more advantageous to the body in the functions Compressed Air: 30 Psi. associated with the mineral. It has been

Kalu et al.; IJBCRR, 24(4): 1-7, 2018; Article no.IJBCRR.34900

suggested that commonly consumed leafy essential amino acid (45.94%) but has higher vegetables are superior sources of calcium total aromatic amino acid (10.44%) than total compared to milk [7]. Deficiency of calcium can sulphur containing amino acid (5.56%). lead to malformation of bones and shells. Melanthera biflora is rich in essential amino Magnesium and potassium are needed for acids, leucine, isoleucine, valine, phenylalanine, maintaining the acid – base balance in the body; lysine, histidine and threonine and can meet the magnesium is an obligate cofactor for DNA minimum daily requirements for them [9]. The synthesis, while potassium is required for muscle dominant essential amino acid is leucine and nerves functions. Melanthera biflora is rich in (11.32%) and is higher than that of G. latifolium magnesium, and so, can be used to supplement (2.25%) and Vernonia amydalina (1.66%) [10]. A low magnesium based stable foods such as 100g serving of Melanthera biflora can provide cassava. It is therefore recommended for about 43-99% of therecommended dietary hypertensive patients, since magnesium has a allowance (Table 5). This high protein content hypotensive effect. Iron content of Melanthera implies that this plant can contributessignificantly biflora was higher than those of T. occidentalis to the daily human protein requirements, usually and A. hybridus [7], and would be beneficial for about 23-56 g. In comparison to the egg protein anaemic patients. Therefore Melanthera biflora and human milk protein reference, the green can serve as an excellent source of minerals for leafy vegetable score higher than those human nutrition. previously reported for G. latifolium and V. amydalina [10]. Table 1. Mineral composition of Melanthera biflora leaves Table 2. Comparison of mineral composition of Melanthera biflora leaves with the WHO Minerals Amount % RDA (mg/100 g) Composition

Iron 14.67 3.829 Copper 2.322 0.608 Minerals Melantera RDA Cobalt 0.041 0.011 biflora Manganese 5.125 1.342 (mg/100 g) Calcium 93.516 24.481 Iron 14.627 15 mg Magnesium 16.17 33.029 Copper 2.322 2 mg Potassium 71.39 18.690 Cobalt 0.041 Less than 20 mg Sodium 32.373 8.476 Manganese 5.125 5 mg Phosphorus 36.50 9.555 Calcium 93.516 1000 mg Selenium 0.006 0.0016 Magnesium 126.171 350 mg Total 381.999 Potassium 71.39 3500 mg Sodium 32.373 2400 mg The Melanthera biflora is rich in ascorbic acid, Phosphorus 36.50 1000 mg which is higher than the concentration previously Selenium 0.006 35 μg reported for some other stable vegetables such End Note: Percentages are based on the weight of the T. occidentalis, T. triangular, C. argentea, S. compounds per the total extract of its family nodiflorium, B. alba, S. acthiopica, A. caudatus, A. hybridus, C. pepo and Amaratus Spinosus (6). Table 3. The vitamin composition of The body cannot synthesis ascorbic acid, so it Melanthera biflora leaves must be obtained entirely through diet, in this connection, Melanthera biflora should serve as a Vitamins Amount % Composition good source of the water soluble vitamin. (mg/100 g) Ascorbic acid is essential for the healthy Vitamin B3 150.3 0.324 formation of bones and teeth. It is a powerful Vitamin B5 0.003 0.065 Vitamin B6 0.0072 0.016 antioxidant [8] whose deficiency results in scurvy Vitamin C 46107 99.37 with swellings of the joints and gums, loosening Vitamin A 0.0038 0.0082 of the teeth and haemorrhages of the skin and Vitamin B1 0.0035 0.0075 mucus membrane. Niacin, thiamine and Vitamin B2 0.0085 0.0018 riboflavin play important roles in nutrient Vitamin D 0.0038 0.0082 metabolism, Melanthera biflora is also a good Vitamin E 0.0085 0.0085 source of these micronutrients. Vitamin B9 0.0077 0.0017 (folic acid) Melanthera biflora leaf contain higher total Vitamin K 0.012 0.026 essential amino acid (47.5%) than total non- Vitamin B12 0.0015 0.0032

Kalu et al.; IJBCRR, 24(4): 1-7, 2018; Article no.IJBCRR.34900

Table 4. Comparison of vitamin from leaves of Melanthera biflora with WHO recommended daily allowance

Vitamins Melanthera biflora (mg/100 g) RDA

Vitamin B3 0.324 20 mg

Vitamin B4 0.065 1.2 mg

Vitamin B6 0.16 1-2 mg Vitamin C 99.37 75 mg Vitamin A 0.0082 600 mg

Vitamin B1 0.0075 1.4 mg

Vitamin B2 0.0018 1.5 mg Vitamin D 0.0082 10 μg Vitamin E 0.0085 8-10 mg

Vitamin B9 (folate) 0.0017 40 μg Vitamin K 0.026 50-100 mg

Vitamin B12 0.0032 1-2 μg

Table 5. Amino acid composition of Melanthera biflora leaves

Amino acid Amount (x 101) (mg/100 g) % Composition Glycine 0.239 3.40 Alanine 0.390 5.55 Serine 0.233 3.32 Proline 0.275 3.92 Valine 0.382 5.44 Threonine 0.410 5.84 Isoleucine 0.414 5.90 Leucine 0.795 11.32 Aspartate 0.936 13.33 Lysine 0.370 5.27 Methionine 0.087 1.25 Glutamate 1.007 14.34 Phenylalanine 0.411 5.85 Histidine 0.187 2.66 Arginine 0.465 6.62 Tyrosine 0.320 4.56 Cystine 0.101 1.44 Total 70.22

Table 6. Comparison of essential amino acid and composition of Melanthera biflora with World Health Organization Recommend daily amount for essential amino acid

Amino acid Melanthera biflora mg/kg boiled mg/70 kg mg/100 kg Histidine 18.7 10 700 1,000 Isoleucine 41.4 20 1,400 2,000 Leucine 79.5 39 2,730 3,900 Lysine 37.0 30 2,100 3,000 Methionine 0.86 45 (total) 1,050 1,500 phenylalanine 41 25 (total) 1,750 2,500 Threonine 41 15 1,050 1,500 Tryptophan - 4 280 400 Valine 38.2 26 1,820 2600 Cysteine 1-0.07 - - -

Kalu et al.; IJBCRR, 24(4): 1-7, 2018; Article no.IJBCRR.34900

Table 7. Comparison of chemical scores of Melenthera biflora leaves with WHO reference protein pattern

Amino acid Reference pattern mg/g protein amino acid score. (%) Human milk (%) Whole egg mg/g A B Glycine 22 + 2 2.89 82.3 15.45 Alanine 46 +2 5.49 71.00 13.88 Serine 61+ 4 6.07 38.4 5.44 Proline 95+5 2.92 94.2 4.13 Valine 51+2 6.02 63.5 10.67 Threonine 44+1 3.41 120.6 13.27 Isoleucine 53+3 50.00 8.3 11.13 Leucine 104+3 6.80 116.9 10.88 Aspartate 86+9 6.09 155.7 15.5 Lysine 71+6 4.64 34.6 2.64 Methionine 61+09 8.01 34.6 2.64 Glutamate 190+8 10.89 93.2 7.55 Phenylalanine 37+1 4.94 83.2 15.8 Histidine 23+2 1.67 111.9 11.57 Arginine 36+3 4.54 102.4 18.39 Tyrosine 46+2 3.21 97.7 9.91 Cystine 20+2.6 1.88 53.7 7.2 A = Melanthera biflora compared with human milk B = Melanthera biflora compared fowl egg protein pattern

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