INTRODUCTION:

Functional foods provide added functional and specific health benefits to consumers' well-being. Thus nowadays many food producers are devoting their time, resources and efforts in the development of these nutraceuticalproducts (functional foods). There are many plants which provide promising health benefits.These plants have bioactive properties, useful minerals and has its own nutrient composition besides having pharmacologically important phytochemicals, thus proving beneficial for human and animal consumption.¹

Many efforts are currently going on in the food industry to develop and market different food supplements, nutraceuticals and associated products as these are the highly growing segments of the food industry.

Functional foods can be considered as a part or borderline to these products they have additional health or physiological benefits over and above the normal nutritional value they provide.²

These nutrients are beneficial and required for the proper physiological functions of the human body. Such nutrients and biochemicals like fats, carbohydrates, proteins play an important role in nourishing human needs for energy and life processes.

Thus the biochemical and nutritional profiling of these plants will be useful and can help to understand the worth of these plants species. Extensive research on determining proximate and micronutrients analysis of various herbal formulations is the need of the hour.WHO also emphasizes on effective standardization of such herbal formulations.

Proximate and micronutrient analysis of the medicinal plants species, viz., Cynodon dactylon (L.) Pers., Annona squamosa Linn., Moringa oleifera Lam, Ocimum tenuiflorum Linn. and Terminalia catappa Linn.was performed in the present study. The total carbohydrates, reducing sugars, proteins and fats were analyzed and the micronutrient content i.e. minerals like Calcium (Ca), Phosphorus (P), Iron (Fe) and Magnesium (Mg) levels were analysed, so also both water soluble and water insoluble vitamins were estimated using biochemical methods.

MATERIAL AND METHODS:

The assays for the estimation of proximate principles and mineral estimation were performed using standardized protocols.³

1) Analysis of proximate principles of diet:

a) Determination of Total Carbohydrates by Anthrone Method^3,4

b) Determination of Total Reducing Sugars by Folin – Wu Method^3,5,6,7

c) Determination of Reducing Sugar (Glucose) by Nelson-Somogyi Method^3,4

d) Estimation of Cellulose Content^3,4

e) Estimation of Crude Fibre Content^3,4

f) Estimation of Total Protein content by Folin Lowry’s Method^3,4,8,9

g) Estimation of Total Free Amino acids Content^3,4

h) Extraction of Total Lipid Content^3,8,10

i) Estimation of Free Fatty acids Content^3,4

2) Micro-nutrient analysis:

Mineral Estimation:

Preparation of sample for mineral analysis:

The ashing method was employed to decompose the organic compounds in the sample by incineration at high temperatures (500⁰C-600⁰C) for 4-12 hr using a muffle furnace. The minerals were then collected from the ash, usually dilute HCL, filtered and diluted to a known volume with deionized water and estimated quantitatively.

a) Estimation of Minerals:

i) Estimation of Calcium by EDTA Method^3,11

ii) Estimation of Phosphorus by Fiske-Subbarow (ANSA) method^3,8,12

iii) Estimation of Iron by Wong’s (KCNS) method^3,13,14

iv) Estimation of Magnesium by Titan Yellow method^3,15,16

b) Estimation of Vitamins:

i) Estimationof Thiamine by Thiochrome method^[3,4]

ii) Estimation of Riboflavin^3,4

iii) Estimation of Niacin by Cyanogen bromide method^3,4

iv) Estimation of Ascorbic acid (Vitamin C) by DNPH method^3,4

v) Estimation of Retinol (Vitamin A) by Carr-Price method^3,11,17,18

vi) Estimation of Tocopherol (Vitamin E)^{3,11, 19}

RESULTS AND DISCUSSION:

The biochemical analysis results of the various nutritional parameters, viz., proximate principles and micronutrients are represented in table 1 and table 2, respectively.

Table 1: Proximate principles of leaf extracts of all the five plants.

Sr. No.	Biochemical parameters	Values of *Moringa oleifera* Lam. (g/100g drywt.)	Values of *Annona squamosa* Linn. (g/100g drywt.)	Values of *Cynodon dactylon* Linn. (g/100g dry wt.)	Values of *Ocimum tenuiflorum* Linn. (g/100g dry wt.)	Values of *Terminalia catappa* Linn. (g/100g dry wt.)
Proximate Principles
1.	Total Carbohydrate content	9.056 + 0.14	12.66 + 0.21	14.64 + 0.23	4.70 + 0.13	17.85+ 0.18
2.	Total Reducing sugar content	3.495 + 0.11	2.66 + 0.12	1.165 + 0.10	0.90 + 0.11	2.594+ 0.13
3.	Total Glucose content	1.924 + 0.12	1.759 + 0.11	0.462 + 0.09	0.405+ 0.10	0.955+ 0.08
4.	Total Cellulose content	11.19 + 0.21	9.569 + 0.19	19.99 + 0.23	1.8 + 0.18	20.10+ 0.16
5.	Total Crude fibre content	9.52 + 0.31	10.67 + 0.29	20.81 + 0.16	2.10 + 0.14	12.07 ±0.59
6.	Total protein content	2.5 + 0.19	23.5 + 0.21	0.825 + 0.10	2.04 + 0.13	9.70+ 0.13
7.	Total Free amino acids content	0.1198 + 0.53	0.89 + 0.85	0.1297 + 0.64	0.102+ 0.70	0.679+ 0.59
8.	Total Lipid/Fat content	1.82 + 0.33	1.56 + 0.28	0.79 + 0.11	0.482+ 0.12	0.745+ 0.18
9.	Total Free fatty acid content	0.19 + 0.06	0.10 + 0.03	0.20 + 0.08	0.05+ 0.02	0.110+ 0.05

*All values are expressed as mean±SD for three determinations

Table 2: Micronutrients of all the five leaf extracts

Sr. No.	Biochemical parameters	*Values of Moringa oleifera* Lam. (mg/100g dry wt.)**	*Values of Annona squamosa* Linn. (mg/100g dry wt.)**	*Values of Cynodon dactylon* Linn. (mg/100g dry wt.)**	*Values of Ocimum tenuiflorum* Linn. (mg/100g dry wt.)**	*Values of Terminalia catappa* Linn. (mg/100g dry wt.)**
Micronutrients
1.	Total Calcium content	480.96 + 0.27	320.64 + 0.19	240.48 + 0.23	180 + 0.17	102.2+ 0.18
2.	Total Phosphorus content	14 + 0.12	8 + 0.12	7 + 0.13	5.62+ 0.20	11.7+ 0.14
3.	Total Iron content	34.43+ 0.10	26.02 + 0.12	32.07+ 0.13	5.72+ 0.19	13.2+ 0.18
4.	Total Magnesium content	1416.0 + 28.64	1875.0 + 24.52	1081.5 + 23.77	5320+ 26.20	3344+ 21.10
5.	Total Thiamin content	0.0187 + 0.0028	0.0187 + 0.0029	0.0196 + 0.0025	0.023+ 0.0022	0.0169+ 0.0019
6.	Total Riboflavin content	0.092 + 0.022	0.085 + 0.017	0.05 + 0.009	0.069+ 0.010	0.079+ 0.018
7.	Total Niacin content	0.290 + 0.019	0.622 + 0.022	0.667 + 0.037	0.896+ 0.23	0.502+ 0.26
8.	Total Ascorbic acid content	516.66 + 76.98	408.33 + 45.31	116.67 + 17.89	72.90+ 14.20	98.20+ 16.89
9.	Total Retinol content	110.41+ 7.54	66.67 + 3.82	50.00 + 2.63	61.24+ 4.01	84.05+ 5.23
10.	Total Tocopherol content	39.545 + 4.87	19.772 + 2.93	24.715 + 3.45	13.25+ 2.40	27.23+ 3.98

*All values are expressed as mean ± SD for three determinations

Today a numerous of the herbal products are used in various herbal formulations which are administered orally, hence the knowledge of proximate and nutrient analysis of these products and raw materials used in it plays a crucial role in assessing their nutritional significance and health effects.^20,21,22

The biochemical analysis of the leaf extracts of three medicinal plants under study,viz., Cynodond actylon (L.) Pers.(CDE), Annona squamosa Linn. (ASE), Moringa oleifera Lam. (MOE), Ocimum tenuiflorum Linn. (OCT) and Terminalia catappa Linn. (TMC) showed considerably high levels of most of the estimated nutritional elements.

Looking at theresults obtained from carbohydrate analysis, CDE showed high levels of total carbohydrates (14.64+0.23g/100g dry wt.), cellulose (19.99+0.23 g/100g dry wt.) and crude fibre (20.81+0.16 g/100g dry wt.) but very less amount of reducing sugars (1.165+ 0.10g/100g dry wt.) and free glucose (0.462+0.09 g/100g dry wt.). Similarly TMC also showed high levels of total carbohydrates (17.85+0.18g/100g dry wt.)

However, the reducing sugars and free glucose content of MOE were (3.495+0.11g/100g dry wt.) and (1.924+ 0.12 g/100g dry wt.) respectivelyand ASE were (2.66+ 0.12g/100g dry wt.) and(1.759+0.11g/100g dry wt.) respectively, was seen to be much higher.

While analyzing the proteins and free amino acids content in the leaf powders of three medicinal plants under study, CDE showed to be very poor sources of proteins and free amino acids.MOE showed considerable amounts of both the proteins (2.5+0.19 g/100g dry wt.) and free amino acids (0.1198+0.53 g/100g dry wt.). Similarly OCT and TMC showed considerable amount of proteins(2.04+0.13g/100g dry wt.) and (9.70+0.13g/100g dry wt.) respectively and free amino acids (0.102+ 0.70g/100g dry wt.) and (0.679+ 0.59g/100g dry wt.) respectively. However, ASE showed maximum amounts of both proteins (23.5+0.21 g/100g dry wt.) and free amino acids (0.89+0.85 g/100g dry wt.).

The results of fat analysis showed that the lipid content was comparatively high in MOE (1.82+0.33g/100g dry wt.) and ASE (1.56+0.28g/100g dry wt.). CDE was found to have low levels of both total lipids (0.79+0.11 g/100g dry wt.) and free fatty acids (0.20+0.08 g/100g dry wt.). Lipid content was found to be (0.482 + 0.12 g/100g dry wt.) and (0.745+0.18 g/100g dry wt.) in OCT and TCM respectively.

Minerals are necessary to life as living organisms use minerals to activate enzymes, hormones and other organic molecules that participate in the growth, functions and maintenance of various life processes. Minerals are an essential part of plants and animals, performing different functions and are used in the synthesis of chlorophyll, proteins and phosphate components of DNA and RNA and energy carrying coenzymes such as ATP.

Minerals cannot be synthesized in the body and are needed to be supplied through plants, vegetables, or mineral rich water. Minerals are the catalysts, which provide a healthy environment in which the body uses vitamins, proteins, carbohydrates and fats. The complete spectrum of minerals is thereby necessary for exemplary health.²³

The micronutrients analysis of the leaf powders of the five medicinal plants under study, showed significant variation among different micronutrients. Magnesium content was found to be the highest followed by calcium in all the five plant leaf powders under study, as compared to the rest of the tested minerals. However, iron and phosphorus contents were found to be comparatively less. Phosphorus content was found to be the least among all the tested minerals in all the five plant leaf powders. MOE showed to be a good source of all the four minerals (calcium, phosphorus, iron and magnesium) as they were found to be in higher amounts in MOE - Calcium (480.96+0.27mg/100g dry wt.), phosphorus (14+0.33mg/100g dry wt.), iron (34.43+ 0.10 mg/100g dry wt.) and magnesium (1416.0 + 28.64 mg/100g dry wt.). However, CDE showed comparatively high iron content (32.07+0.13 mg/100g dry wt.) andASE comparatively high levels of magnesium (1875.0+24.52mg/100g dry wt.). OCT showed maximum levels of magnesium (5320.0+26.20 mg/100g dry wt.).

Most of the vitamins were found to be in good quantity especially in ASE,MOE, OCT and TMC; with the exception of niacin which was found to be in less quantity in MOE (0.290+0.019mg/100g dry wt.)

Aslam et al., (2005)²³, in their study, have determined the mineral composition of Moringa oleifera Lam. leaves and other parts like pods from different regions of Punjab, Pakistan. The result of their study has shown that the pods and leaves of Moringa oleifera Lam., indigenous to different agro-climatic regions of Punjab, contained a considerably high amount of Calcium, Magnesium, Potassium, Manganese, Phosphorus, Zinc, Sodium, Copper and Iron. Hence it was concluded by Aslam et al.,(2005)²³ that the pods and leaves of Moringa oleifera Lam. might be used as a feasible supplement of dietary minerals. The results of our study are found to be in accordance with the report of Aslamet al.,(2005)²³ andindicated that the Moringa oleifera Lam. leaf extract was found to be a good source of all the four minerals (Calcium, Phosphorus, Iron and Magnesium) as they were found to be in higher amounts in MOE.

Other studies by Bernardo et al.,(2013)²⁴and Mousavi et al., (2019)^[25] have demonstrated the similar kind of results for the leaf extract of Ocimum spps. (basil) for the parameters such as carbohydrates, crude protein, crude fibre, minerals and vitamins content.

However, no proper previous study reports were found on the nutritional/biochemical analysis of the leaf powders of Cynodond actylon (L.) Pers., Annona squamosa Linn. And Terminalia catappa Linn. indicating their important nutrients, minerals and vitamins content. So also the essential nutrients (carbohydrates, lipids and proteins) and vitamins levels of Moringa oleifera Lam. leaves were not found to be previously reported, hence our study may be considered as the first to report the same.

CONCLUSION:

The evaluation of various proximate parameters for the leaves of Cynodond actylon (L.) Pers., Annona squamosa Linn., Moringa oleifera Lam., Ocimum tenuiflorum Linn. And Terminalia catappa Linn. Has given an idea about the specific characteristics of these crude drugs under examination, in their powder form.

Biochemical analysis of the extracts of five medicinal plants leaves showed considerably high levels of most of the estimated nutritional elements. The powders of the plant leaves were found to be having all the essential nutrients, minerals and vitamins in considerable amounts and possess good nutritive value. Hence, it can be concluded that the leaf powders of Cynodond actylon (L.) Pers., Annona squamosa Linn., Moringa oleifera Lam., Ocimum tenuiflorum Linn. And Terminalia catappa Linn. can be looked forward as the probable sources of food supplementation and medical foods in future after further investigation of the anti-nutritive factors present in them and their enzymatic and molecular effect on human health.

ACKNOWLEDGEMENT:

Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.

CONFLICTS OF INTEREST:

The authors declare no competing interests.

AUTHORS’ CONTRIBUTION:

All the authors of this paper equally contributed towards the experimental work as well as the writing of this paper.

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Received on 20.03.2022 Modified on 23.06.2022

Research J. Pharm. and Tech 2023; 16(8):3680-3684.

DOI: 10.52711/0974-360X.2023.00605