Nutritional Value and Elemental Analysis of Katphala ( Myrica esculenta Buch-Ham)

Atul Kabra1* & Uttam Singh Baghel2

1 IKG Punjab Technical University, Kapurthala, Punjab, INDIA

2 Kota College of Pharmacy, Kota, Rajasthan, , INDIA

* Correspondence: E-mail:

(Received 04 May, 2018; Accepted 04 August, 2018; Published 08 August, 2018 )

ABSTRACT: Myrica esculenta (Family: Myricaceae) commonly known as Kaiphala or Katphala is a widely used medicinal plant in Ayurveda. Traditionally its leaves are used in the treatment of fever, catarrh of mucous membrane, asthma, diarrhoea and bronchitis, its paste is applied to cuts and wounds . In spite of its numerous medicinal attributes, no published work is available till date on nutritional values and elemental analysis of leaves. The nutrient and antinutrient compositions of the leaves were investigated and compare the evaluated data with reported nutritional parameters of fruit. The result of proximate analysis shows that Myrica fruit had higher moisture content, crude lipids and carbohydrate contents values of 72.33±0.23, 4.93±0.06 %and 78.03±0.14 % while those of the leaves had 3.54±0.11 %, 1.38±0.54 % and 46.19±0.21 %. The total ash, crude fibre and crude protein contents in the leaves were higher values of 8.3±0.28 %, 22.45±0.17 % and 10.55±0.22 % while the fruit had 2.18±0.02 %, 5.22±0.08 % and 9.62±0.03 %. The antinutrient parameters evaluated in the leaves were phytic acid (2.46±0.16) %, saponin content (1.76±0.25 %), alkaloidal contents (0.28±0.06) and oxalate content (0.27 ± 0.10) %. The predominant mineral elements in the leaf powder according to ICP-MS were Ca (42.59 mg/100g), Mg (14.82 mg/100g), and Na (12.41 mg/100g). Lead and arsenic contents were not detected in the leaves and fruits of Myrica. Hence the outcome of this study revealed that M. esculenta leaves could be a valuable nutraceutical supplement and a cheapest source of essential nutrients to the human diet.

Keywords: Macronutrients; proximate analysis; elemental analysis; oxalate content and ICP-MS.

INTRODUCTION: Plants are used as medicine to maintain human health from ages1 and are also major natural sources of medicinal compounds in current pharmacopoeias.2 The World Health Organization reports that up to 80% of people still rely primarily on traditional medicines.3 The medicinal value of these plants due to the presence of nutrients, minerals and a variety of phytochemicals. These nutrients and minerals are essential for physiological functions of human body. Nutrients and biochemical compounds such as carbohydrate, protein and lipid play an important role in satisfying human needs for energy and life process 4.

Myrica esculenta Buch.-Ham. syn. Myrica nagi F. (Myricaceae), commonly known as ‘Box Berry’, ‘Kaiphal’, ‘Katphala’ is an important medicinal tree distributed all along outer Himalaya from Ravi (Punjab) eastwards to Assam, Khasia, Jaintia, Shimla, Bengal, Naga and Lushai hills at altitudes of 900-2100m. M. esculenta is a small to moderate sized, evergreen, dioecious tree indigenous to subtropical temperate region.5-9 Trees of Myrica grow well in nitrogen depleted soils and are common associates of Pine (Pinus sp.) and Oak ( Quercus leucotrihophora). They are also found in mixed forests and in agricultural and marginal lands.10 M. esculenta known for edible fruit and other by-products is a potential income-generating species in the sub-Himalayan region.11 As per the Ayurvedic literature, various parts of this plant is used in gulma (abdominal tumors), jvara (fever), arsa (piles), grahani (irregular bowel function), pandu roga (anemia), hrillasa (nausea), mukha roga (oral disorders), kasa (cough), svasa (dyspnea), agnimandhya (indigestion), aruchi (anorexia) and kantharoga (ears, nose, and throat disorders). 12

Traditionally, leaves of this plant are useful remedy in fever, catarrh of mucous membrane, asthma, diarrhoea and bronchitis.13 Leaf paste is applied to cuts and wounds.14 Bark is widely used in the Indian systems of medicine as astringent carminative, antiseptic and in the disease supposed to be caused by deranged phlegm, such as catarrhal, fever, cough and affection of throat.15 Paste of the bark is applied on the chest to get relief from cough & bronchitis.16 Fruits are utilized in food industries in Himalayas in different forms like syrups, jam, and squash.17 Due to the high medicinal values of the leaves, bark and fruits of this medicinally important tree are imported and exported.18

Several valuable reviews of the ethnobotanical uses and phytochemistry of M. esculenta are available. Myrica is found to be a rich source of phenolic compounds, triterpenoids, saponins, glycosides. flavonoids and flavonols.19 The leaves, in particular, have been found to contain flavonoids, tannins, diarylheptanoids, phenolic acids and flavonoids; these compounds have various biological activities, including antihypertensive, anti-inflammatory, antidiabetic, antihelmintic by using a number of in vitro and in vivo animal models, which prove the traditional utilization of leaves scientifically.20

The plant has tremendous therapeutic potential with every part of the plant being used medicinally. Despite vast medicinal uses of leaves, no report is known about its nutritional analysis hence the aim of this study were to determine the proximate nutritional value and elemental compositions of M. esculenta leaves and also to providing scientific data based on our findings in relation to its dietary and medicinal applications.


Collection and authentication of plant materials: Leaves of Myrica esculenta used for this study were collected from out skirt of Chail Chowk, Mandi, Himachal Pradesh (H.P.) in November 2016. The plant was identified, authenticated and certified (AGI/16/1245) by Prof. Suresh Kumar, Head Department of Botany, Abhilashi Group of Institutions, Mandi, H.P. Freshly collected plant material was cleaned to removed adhering dust and then dried under shade. The dried sample groundedinto powder and kept in air tight container for further studies.

Reagents: All chemicals used for the experiment were of analytical grade (Merck, Germany).

Proximate analysis: Powdered leaf sample were dried in an oven at 110oC for 1hr to a constant weight for moisture determination. Ash content, crude fibre, crude protein and crude lipid were analyzed by triplicate according to AOAC method.21 The total carbohydrates were determined by the subtraction method [100 - (% crude proteins + % crude lipids + % moisture content + % ash)].22 The energy values (kcal/100g) were determined by multiplying the values of proteins, lipids, and carbohydrates by factors of 4, 9, and 4, respectively, and taking the sum expressed in kilocalorie .23

Anti-nutrient analysis:

Determination of oxalate content: For the determination of oxlate content modified titration method was used. 24, 25 One gm of pulverized sample was placed in conical flask. 75 mL of 3M H2SO4 was added. The solution was carefully stirred with a magnetic stirrer for 1 hour and then filtered using Whattman filter paper No.1. 25 mL of the filtrate was collected titrated against 0.1M KMnO4 solution till a light pink colour that persisted for 15 seconds appeared. The oxalate content was calculated by taking 1 mL of 0.05 mol/L of KMnO4 as equivalent to 2.2 mg oxalate.

Determination of phytic acid: The phytic acid content was determined as described method.26 2.0 g of sample was weighed into a 250 mL flask, 100 mL of 2% concentrated HCl was added and allowed to stand for 3 hrs and then filtered with Whattman filter paper No.1. 25 mL of the filtrate was placed in 250 mL of conical flask with 10 ml of 0.3% ammonium thiocyanate solution as indicator and titrated with standard Iron III Chloride solution containing 0.00195 g Iron/mL, end point observed to be brownish yellow colour persisted for 5 min. The percentage phytic acid was calculated as:

Phytic acid (%) = titre value × 0.001 95 × 1.19 × 100

Determination of alkaloidal content: Alkaloid content was determined as described method.27 5 g of plant extract was added with 200 mL of 10% acetic acid in ethyl alcohol and mixed. The mixture was covered and allowed to stand for 4 hrs. and then mixture was filtered with Whattman filter paper No.1. Filtrate was concentrated to a 1/4th of its original volume on a water bath. Concentrated ammonium hydroxide was added in drops to the extract until precipitation was completed. The solution was allowed to settle, washed with dilute NaOH solution and then filtered. The residue collected was dried and weighed. The alkaloid content was calculated using the equation:


Determination of saponin content: Saponin content was estimated as described by Obadoni and Ochuko. 28 5 g of the plant sample was added to 50 mL of 20% ethyl alcohol, kept on a shaker for 30 min after that heated in a water bath for 4 h at 55ºC. The resulting mixture was filtered and the residue re-extracted with another 200 mL of 20% aqueous ethanol. The filtrates were combined and reduced to 40 mL in a water bath at 90oC. The concentrate was transferred into a separating funnel, 20 mL of diethyl ether was added, and shaken vigorously. The upper ether layer was discarded and the aqueous (bottom) layer retained in a beaker. The retained layer was re-introduced into a separating funnel and 60 mL of n-butanol was added and shaken vigorously. Butanol extract present in the upper layer was retained while the bottom layer was discarded. The butanol layer was washed twice with 10 mL of 5% aqueous NaCl. The remaining solution was collected and heated to evaporation in a water bath, then dried to constant weight at 40 oC in an oven. The saponin content was calculated using the equation:


Elemental analysis: Leaf samples were digested in concentrated HNO3. The digest was transferred to a 25-ml volumetric flask, and the volume was adjusted to 25 ml with deionized water. A blank digest was prepared in a similar manner. The multi elemental analysis was performed by inductively coupled plasma mass spectrometry (ICP-MS; XSeries 2, Thermo Scientific).

Statistical analysis of data: The results were statistically analyzed and expressed as mean (n = 3) ± standard deviation. All experiments were performed in triplicates and the data expressed as mean ± SD using the Microsoft Excel 2010 spreadsheet.


Proximate composition: The results of proximate analysis of leaves and fruits of M. esculenta were shown in Table 1. There were a significant difference in the total ash and crude fiber contents in the leaves ad fruits of Myrica with the leaves having the higher values of 8.3±0.28 and 22.45±0.17 %. Sood et al. reported that moisture content and carbohydrate contents having significantly higher value than leaves were 72.33±0.23 % and 78.03±0.14 %.20 The lipid content was also significantly high in the fruits than leaves was 4.93±0.06 %. The total energy derived from this plant leaves and fruits calculated were 239.49±0.080 and 394.98±0.012 %.

Anti-nutrient composition: The summary of anti-nutrient composition of the sample is presented in Table 2. Four anti-nutrient were studied, phytic acid content was most abundant with a percentage value of 2.46±0.16 %, which was closely followed by saponin 1.76±0.25 %. The alkaloid and oxalate content were least values 0.28± .06 % and 0.27±0.10%. There is no data reported on the antinutrient analysis of fruits.

Table 1: Proximate nutritional composition of dried M. esculenta leaf and fruit.


Composition %



Moister content



Total ash



Crude lipid



Crude fibre



Crude protein






Energy value

(Kcal/100 gm)



Values expressed as Mean ± SD, n=3

Statistically significant differences between the means of both are denoted as

** p < 0.001; * p < 0.05.

Table 2: Anti-nutrient Composition of M. esculenta leaf.


Values %

Phytic acid








Values expressed as Mean ± SD, n=3 on dry

weight basis.

Mineral Composition: Table 3 represents the mineral composition of both the leaves and fruits of Myrica. Calcium, magnesium, sodium, manganese, zinc, and iron contents were significantly higher in the leaves than fruits. Their values were 42.59 mg/100g, 14.82 mg/100g, 12.41 mg/100g, 3.45 mg/100g, 2.32 mg/100g, and 1.71 mg/100g. Potassium content was reported slightly higher value in the fruits than leaves 6.86 mg/100g. Arsenic and lead contents were not detected in the leaves and fruits both.

The shelf-life and stability of any food component is determined by its moisture content. The low moisture content indicates that food products have a long shelf life and reduced microbial contamination.29 The ash content evaluated from this leaves was relatively indicating that its richness in mineral nutrients. These minerals act as inorganic co-factors in metabolic processes.30 The percentage of dietary fibre content was relatively high of the sample. The relatively high fibre content prevents constipation by facilitates peristaltic movement and aids the absorption of certain minerals in the gut and reduces cholesterol absorption.31, 32 High fibre intake could lead to a decline the incidence of diseases associated with metabolic disorders.33 The lipid content in the sample was very less and it was lowest nutritional composition evaluated in this study. Dietary lipids enhance the taste of food by preserving its flavours.34 Excess intake of dietary lipids increases the risk of cardiovascular diseases, aging, and cancer. In this concern, the low crude lipids content of M. esculenta leaves indicates that it could prevent certain chronic diseases associated with lipids in human beings. The crude protein content in the sample was third highest proximate composition of the sample

Table 3: Measured mineral composition of M. esculenta leaf and fruit (mg/100 g).


mg/100 g DW




































Values expressed as Mean ± SD, n=3 on dry weight basis.

ND: non determined

Statistically significant differences between the means of both are denoted as

** p < 0.001; * p < 0.05.

The relatively high protein level of M. esculenta leaves could make it a useful supplement to diets. Dietary proteins play an important role in the manufacturing and safe guarding of certain organic materials necessary for the smooth functioning of the human body.35 Proteins also serves the purpose of enzymatic catalyst and mediates metabolic and energy regulation.36 Carbohydrate content was the highest nutritional composition. The high carbohydrate content makes it rich source of energy and this could be used to enhance the energy content of diets.34 The overall energy derived from M. esculenta leaves sample calculated was 239.49±0.080 kcal/100g which is below the recommended daily energy value. Therefore M. esculenta as a low energy food source maybe very helpful in weight management programmes as used by traditional practioners.

The saponin content in M. esculenta was low and within the safe limit, since an amount below 10% is not hazardous to the body.37 High saponin levels in human and animal diets have been implicated in growth impairment; decrease the bioavailability of nutrients and inhibition of biochemical reactions that promote breakdown of ingested proteins. 38 Alkaloids are one of the most efficient therapeutic bioactive components in plants. For instance, consumption of high tropane alkaloids will cause rapid heartbeat, paralysis and in fatal case, lead to death. Uptake of a high dose of tryptamine alkaloids will lead to the staggering gait and death.39 Other toxic effect of alkaloids includes disruption of the mucus membrane in the gastrointestinal tract. 40 The alkaloid content recorded in this study was quite low, attenuating the fear of anti-nutrient activity. The phytate content of M. esculenta leaf sample was low. Phytate chelates with metal ion such as calcium, copper, zinc, magnesium and iron to form an insoluble complex that are not readily absorbed from gastro intestinal tract (GIT).41,42A dietary phytate content of 1% - 6% over a long period decrease zinc bioavailability.43

Oxalate form chelate with toxic metals such as lead and mercury and show antioxidant effect. The presence of oxalate in foods causes irritation in the mouth and could diminishes the absorption of calcium and increase the risk of kidney stone.44, 45 The concentrations of anti-nutrients (saponin, oxalate, phytate and alkaloids) recorded in this study were however within the safe limit and may not elicit toxic effect when consumed.

Minerals are considered to be essential in human nutrition for the overall physical and mental health, as well as important constituents of nerve cells, bones, teeth, tissues, muscles, and blood. Table 3 represents the result of mineral composition available in M. esculenta. The results revealed that leaves of this are a good source of macro and micro both minerals. The minerals evaluated are in the order Ca>Mg>Na> K> Mn>Zn> Fe>P >Cu.

Calcium play a vital role in muscle contraction, neurological function, blood clotting, neurological examination, regulation of cell permeability and osteoporosis. It is also needed for building and strong bones and teeth.46, 47 Thus, M. esculenta is considered to be a natural cure for osteoporosis. Furthermore Magnesium plays an important role in enzyme activity and prevents from heart disease. It also plays an important role in formation and function of bones, muscles, nerve transmission and immune system. Magnesium plays a vital importance in strengthening of ß-cells functions thus preventing onset of diabetes. 48 Sodium is very important mineral element involved in the transmission of nerve impulse as well as maintenance of osmotic pressure of body fluids.49 Deficiency of sodium cause dehydration and muscle cramps.50 Potassium plays an important role in water as well as acid-base balance in the body. It is also responsible for maintaining cardiac rhythm, nerve action and functioning of muscles. Deficiency of potassium cause muscle paralysis.45

Four micro elements (Fe, Cu, Zn, Mn) were present in the sample. Manganese serves as a cofactor of various enzymes involved in metabolism essential for reproduction function, growth, and skeletal development. It also plays an important role in the metabolism of carbohydrates, proteins, cholesterol and also involved in formation of urea.51, 52

Iron is an important constituent of Hemoglobin (Hb) and carries oxygen in the blood. It plays an important role in formation of tendons and ligaments. Deficiency of iron cause anaemia, weakness, poor resistance o infection and in female may cause infertility.53 Therefore, M. esculenta could be used as a potent source to improve the anemic condition of a patient. Zinc a vital traces element found in many enzymes, needed for brain development, normal growth, bone formation, wound healing, sperm production and sexual maturation.54 Therefore this sample can be a good zinc supplement for people with infertility challenges. Copper is a part of various enzymes such as cytochrome oxidase, lysyl oxidase, ceruloplasm required for metabolism of iron in blood, it also involved in erythropoiesis, erythrocyte function and survival. Copper deficiency can cause cardiac disorders, anemia and neutropenia. 50

CONCLUSION: The study revealed that M. esculenta had high crude fibre, ash content, calcium, magnesium, manganese and iron contents. The antinutrient contents were within acceptable limit and may not affect the absorption of other nutrients. M. esculenta is rich in many macro and micro nutrients and can therefore serve as a supplement to prevent many mineral deficiencies. M. esculenta should therefore be considered a plant with great potential in the food, nutritional and pharmaceutical industries. Further studies on toxicity of M. esculenta are on-going to ascertain its possible adverse effects and to confirm some of the ethnopharmacological claims.


1. Upadhya V, Hegde H. V., Bhat S., Hurkadale P. J., Kholkute S. D., and Hegde G. R. (2012) Ethnomedicinal plants used to treat bone fracture from North-Central Western Ghats of India, J. Ethnopharmacol, 142, 557-62.

2. Kingston D. G. (2011) Modern natural products drug discovery and its relevance to biodiversity conservation, J. Nat. Prod., 74, 496-511.

3. Ekor M. (2014) The growing use of herbal medicines: issues relating to adverse reactions and challenges in monitoring safety, Frontiers in Pharmacology, 4, 1–10.

4. Novak W. K., Haslberger A. G. (2000) Substantial equivalence of antinutrients and inherent plant toxins in genetically modified novel food, Food Chem. Toxicol., 38, 473-483.

5. Paranjpe P. (2012) Indian medicinal Plants, (New Delhi, Chaukhamba Sanskrit Pratishthan, 128).

6. The Ayurved Pharmacopoeia of India, Part-I, Vol-II, Govt. Of India, Ministry of Health & Family Welfare, Dept. of AYUSH 2, 90.

7. Annonymous. (1962) The Wealth of India: Raw Material, (New Delhi, Publications and Information Directorate, 472).

8. Parmar C., Kaushal M. K. (1982) Myrica nagiIn: Wild fruits, New Delhi, Kalyani publishers, 49-53).

9. Chauhan N. S. (2006) Medicinal and Aromatic Plants of Himachal Pradesh, (New Delhi, Indus Publishing Company, 267).

10. Bhatt I. D., Rawal R. S., Dhar U. (2000) Improvement in seed germination of Myrica esculenta Buch. Ham. Ex D. Don- A high value tree species of Kumanun Himalaya, India, Seed Sci. Technology, 28, 597-605.

11. Pandey G., Sharma B. D., Hore D. K., Rao N. K. (1993) Indigenous minor fruits genetic resources and their marketing status in north-eastern hills of India, Journal of Hill Research, 6, 1–4.

12. The Ayurvedic Pharmacopoeia of India part I. vol. III, 1st ed. (1999) (Government of India, Ministry of Health and Family Welfare, Department of Indian System of Medicine and Homeopathy, New Delhi, 92-93.

13. Gaur R. D. (1999) Flora of District Garhwal: North West Himalaya, Srinagar (Garhwal), Transmedia, 105.

14. Kichu M., Malewska T., Akter K., Imchen I., Harrington D., Kohen J., Vemulpad S. R. and Jamie J. F. (2015) An ethnobotanical study of medicinal plants of Chungtia village, Nagaland, India. J. Ethnopharmacol, 166, 5-17.

15. Kirtikar K. R., and Basu B. D. (1999) Indian Medicinal Plants. (New Delhi, International book distributors, 1699).

16. Nadkarni K. M. (1954) Indian Materia Medica, (Bombay, Popular book depot, 828-829).

17. Patel K. G., Rao N. J., Gajera V. G., Bhatt P. A., Patel K. V., and Gandhi T. R. (2010) Antiallergic activity of stem bark of Myrica esculenta Buch. Ham. (Myricaceae), J. Young Pharm. , 2(1), 74-78.

18. Nadkarni K. M. (2002) Indian Materia Medica, (Bombay, Popular Prakashan Pvt. Ltd., 871).

19. Gusain Y. S. and Khanduri V. P. (2016) Myrica esculenta wild edible fruit of Indian Himalaya: need a sustainable approach for indigenous utilization, Eco. Env. Cons., 22, S267-S270.

20. Sood P. And Shri R. (2018) A Review on Ethnomedicinal, Phytochemical and Pharmacological Aspects of Myrica esculenta, Indian J. Pharm. Sci., 80(1), 2-13.

21. AOAC M. (1995) International, Official Methods of Analysis. (Arlington VA, USA: Association of Official Analytical Chemists).

22. Muller H. G., and Tobin G. (1980) Nutrition and Food Processing, (London, Croom Helm Croom Helm, 320).

23. Javid H., Farman U. K., Riaz U., Muhammad Z., Rehman N. Y., Shinwari Z. K., Khan I. U., Zohaib M., Din I. U. and Hussain S.M. (2011) Nutrient evaluation and elemental analysis of four selected medicinal plants of Khyber pakhtoon khwa Pakistan, Pakistan J. of Botany, 434, 427–434.

24. Imran M., Khan H., Hassan S. S., and Khan R. (2008) Physicochemical characteristics of various milk samples available in Pakistan,Journal of Zhejiang University Science, 9(7), 546– 551.

25. Aina V. O., Sambo B., Zakari A., Haruna H.M.S., Umar K. Akinboboye R.M., and Mohammed A. (2012) Determination of nutritional and antinutritional content of Vitis vinifera (Grapes) grown in Bomo (Area C) Zaira, Nigeria. Adv. J. Food Technol., 4(6), 225-8.

26. Day R. A., and Underwood A. L. (1986) Quantitative analysis (Prentice Hall Publication, 701).

27. Damilola O. L., Joseph O. B., Olufemi A., and Amoo I. A. (2013) Chemical composition of red and white cocoyam (Colocosia esculenta) leaves, Int. J. Sci. Res., 11, 121-125.

28. Obadoni B. O. and Ochuko P. O. (2001) Phytochemical studies and comparative efficacy of the extracts of some haemostatic plants in Edo and Delta States of Nigeria, Glob. J. Pure Appl. Sci., 8, 203-18.

29. Uyoh E. A., Ita E. E., and Nwofia G. E. (2013) Evaluation of the chemical composition of Tetrapleura tetrapter (Schum and Thonn.) Tuab. accessions from Cross River State, Nigeria, Int J Med Arom Plants, 3, 386-94.

30. Jonathan A. A. and Funmilola A. S. (2014) Nutritional and anti-nutritional composition of Bridelia ferruginea Benth (Euphorbiaceae) stem bark sample, Int. J. Scient. Res. Knowl., 2, 92-104.

31. Ogungbenle H. N., and Omosola S. M. (2015) The comparative assessment of nutritive values of dry nigerian okra (Abelmoschusesculentus) fruit and oil, Int. J. Food Sci. Nutr. Eng., 5, 8-14.

32. Ekwumemgbo P. A., Sallau M. S., Omoniyi K. I., and Zubairu S. Y. (2014) Proximate and anti-nutritional constituents of Abelmoschus esculentus grown in Fadaman Kubanni, Zaria, Kaduna State, Niger. J. Sci. Res. Rep., 3, 2015-27.

33. Ikewuchi C. C., Ikewuchi J. C. (2009) Chemical profile ofPleurotus tuberregium (fr) sing's sclerotia pacific, J. Sci. Technol., 10, 295-9.

34. Anita B. S., Akpan E. J., Okon P. A., Umoren I. U. (2006) Nutritive and antinutritive evaluation of sweet potatoes (Ipomoea batatas) leaves, Pak. J. Nutr., 5, 166-8.

35. Hayat I., Ahmad A., Ahmed A., Khalil S., and Gulfraz M. (2014) Exploring the potential of red kidney beans (Phaseolus vulgaris L.) to develop protein-based product for food applications, J. Anim. Plant Sci., 24, 860-8.

36. Hussain J., Khan A. L., Rehman U. R., Khan F., Hussain S. T., Shinwari Z. K. (2009) Proximate and nutrient investigations of selected medicinal plants species of Pakistan, Pak. J. Nutr., 8, 620-4.

37. Igile G. O., Iwara I. A., Mgbeje B. I. A., Uboh F. E., and Ebong P. E. (2013) Phytochemical, proximate and nutrient composition of Vernonia calvaona Hook (Asterecea): a green-leafy vegetable in Nigeria, J. Food Res., 2(6), 1-10.

38. Das T. K., Banerjee D., Chakraborty D., Pakhira M. C., Shrivastava B, and Kuhad R. C. (2012) Saponin: role in Animal system, Vet World, 5(4), 248-54.

39. Gemede H. F., and Ratta N. (2014) Antinutritional factors in plant foods: potential health benefits and adverse effects, Int. J. Nutr. Food Sci., 3(4), 284-9.

40. Fernando R., Pinto M. D. P., and Pathmeswaran A.(2012) Goitrogenic food and prevalence of goitre in Sri Lanka, J. Food Sci., 41, 1076-81.

41. Egbuna C. and Ifemeje J. C. (2015) Biological functions and anti-nutritional effects of phytochemicals in living system, IOSR J. Pharm. Biol. Sci., 10, 10-19.

42. Akande K. E., Doma U. D., Agu H. O. and Adamu H. M. (2010) Major antinutrients found in plant protein sources: Their effect on nutrition, Pak. J. Nutr., 9, 827-32.

43. Oke O. L. (1969) Chemical studies on the more commonly used vegetables in Nigeria, Afr. Sci. Ass., 11, 42-8.

44. Fekadu H., Beyene F., and Desse G. (2013) Effect of traditional processing methods on nutritional composition and anti-nutritional factors of anchote (Coccinia abyssinica Cogn) tubers grown in Western Ethiopia, J. Food Process Technol., 4, 249.

45. Food and Agricultural Organization of the United Nations (FAO) World Health Organization. Human vitamin and mineral requirements. Report of a Joint FAO. Rome: WHO expert consultation, Bangkok, Thailand; 2001.

46. Haq F., and Ullah R. (2011) Comparative determination of trace elements from Allium sativum, Rheum australe andTerminalia chebula by atomic absorption spectroscopy, Int. J. Biosci., 1, 77-82.

47. McCarron D. A., Kazaks A. G., Geerling J. C., Stern J. S., and Graudal N. A. (2013) Normal range of human dietary sodium intake: a perspective based on 24-hour urinary sodium excretion worldwide, Am. J. Hypertens., 26(10), 1218-23.

48. Michael W. K. (2007) Medical Biochemistry, (London, Queen Mary Publishers, 13-17).

49. Shomar S. (2012) Major and trace elements in Nigella sativa provide a potential mechanism for its healing effects, J. Med. Plants Res., 6(34), 4836-43.

50. Johnson W. T., Lieberman H. R., Kanarek R. B., Prasad C. (2005) Nutritional and neuroscience, (Boca Raton, Taylor & Francis, 17).

51. National Nutrition Surveys of Australia Ministry of Health. NZ food: NZ people. Key results of the 1997 National Nutrition Survey. Wellington: Australia. 1998.

52. Lee S. H., Jouihan H. A., Cooksey R. C., Jones D., Kim H. J., Winge D. R., and Mclain D. A. (2013) Manganese supplementation protects against diet-induced diabetes in wild type mice by enhancing insulin secretion, Endocrinology, 154, 1029-38.

53. Institute of Medicine (2006) Dietary Reference Intakes: The Essential Guide to Nutrient Requirements , (Washington DC (USA), National Academic Press, 1344).

54. Jabeen S., Shah M. T., Khan S., Hayat M. Q. (2010) Determination of major and trace elements in ten important folk therapeutic plants of Haripur basin, Pak. J. Med. Plant Res., 4(7), 559-66.