INTRODUCTION:

Plant and animal meals, and their different components, are the principal sources of sustenance for humans. Nutrition is essential for both human and animal growth and development, as well as illness prevention and treatment. Nutrition is also essential for maintaining excellent health and functionality¹. Science of nutrition helps us make better choices and not only to improve our health but also reduce our risk of disease². Nutrition support and metabolic care are indispensable parts of patient care³. These Nutrients includes macronutrients such as Carbohydrates, proteins, fats, fiber and micronutrients such as iodine, minerals, iron, calcium and vitamins⁴.

These nutrition are used for the various clinical diseases includes cardiovascular disease, tumors, liver injury, diabetes, to increase muscular performance for sports personality, as taste enhancer to improve food quality, to prevent obesity, to improve lactation effect to new mothers, and also used in prevention, prophylaxis and treatment of various disease⁵.

The medicinal potential of natural resources, such as seeds and herbs, for the treatment of human ailments has sparked a lot of attention. Since medicinal plants contain a wide range of chemical substances that have specific physiological effects on humans, they have been utilised for thousands of years to cure a variety of human ailments. Herbs are a great source of medicinal chemicals and have several uses in the pharmaceutical sector⁶. The main aim of the present research is to study nutritional values and phytochemical constituents of Cassia tora, Moringa oleifera, Asparagus racemose which may possess a good amount of various phytochemical activity.

Cassia tora:

Cassia tora, an annual herb, belongs to the family Leguminosae. It is also known as ‘Chakramard’ in Ayurveda, ‘Panwar’ in Unani and ‘Jue Ming Zi’ in Chinese system of medicine⁷. Cassia tora is very tolerant and easily located plant in India, it mainly grows in wasteland rainy season⁸^,⁹. It has immense nutritional and medicinal values. It contains amino acids, proteins, fats, carbohydrates, fibres, vitamins, and minerals⁷. The important secondary metabolites present in C. tora are flavonoids, alkaloids, anthraquinones glycosides, phenols, and saponins¹⁰. The seeds of cassia tora contains the chemical constituents such as Sitosterol, Chryso-obtusin, Stigmasterol, Norrubrofusarrin, Obtusin, Chrysophanol, Physcion, Aurantio-obtusin, Cassiaside, Rubrofusarin etc¹¹. They are reputed in Ayurvedic and Chinese system of medicine to treat night blindness, hypercholesterolemia, hypertension, and constipation¹². Seeds of Cassia tora L. are reported to contribute in the various pharmacological activities including antihepatotoxic, hypolipidemic, antifungal, antimutagenic, and hypoglycemic activity⁷^,¹³.

Figure 1: Chemical constituents of Cassia tora

Moringa oleifera:

Moringa oleifera Lam. is native to the Indian subcontinent and in the tropical and subtropical areas around the world¹⁴. Locally referred to as the "drumstick tree," Moringa oleifera Lam is a member of the Moringaceae family and may be found growing wild in the Indo-Pakistan subcontinent's sub-Himalayan region. It is also farmed across the tropical belt¹⁵^,¹⁶. The leaves of Moringa oleifera composed of chemical constituents such as Lutein, Beta-carotene, Polyprenol, phytyl fatty acid ester, Chlorophyll-a, Beta-sitosterol, Triacylglycerols etc¹⁷^,¹⁸. It also contains abundant amounts of Ca, K, Cu, P, Fe, Mg and S. Therefore, plants may be considered a substitute for addressing hunger in different regions of the world. In addition to being nutrient-rich, it also contains a lot of antioxidants (vergara), and studies have shown that it has anti-diabetic, anti-cancer, anti-microbial, anti-allergic, and antipyretic effects.¹⁹^,²⁰.

Figure 2: Chemical constituents of Moringa oleifera

Asparagus racemosus: Asparagus racemosus Wild. is a woody climber belonging to the family Asparagaceae²¹. They have small uniform needle shaped leaves and flowers are white and have tiny spikes. The roots of Asparagus racemosus composed of chemical constituents such as Sarsasapogenin, Shatavarin-1, Shatavarin-4, Rutin, Adscendin A and B, Asaranin A, Band C etc²². It possesses a number of pharmacological activities such As aphrodisiac, to promote milk secretion, immunostimulant, antioxidant Antihepatotoxic, antibacterial, antidiabetic, Antineoplastic, antidiarrheal, antiulcer, antioxytocic, etc^23,24.

Figure 3: Chemical Constituents of Asparagus racemosus

Malnutrition:

When the body does not get the proper number of vitamins, minerals, and other nutrients (protein) to sustain healthy conditions and organ function, the condition known as malnutrition occurs²⁵. People who are either overfed or undernourished suffer from malnutrition²⁶.

Research conducted by the National Nutrition Monitoring Bureau (NNMB) revealed that over 40 lac youngsters in Maharashtra suffered from grade 2 to 4 malnutrition. An estimated 82,000 children in the state's rural parts, 23,500 in its tribal areas, and 56,000 in its urban slums perished annually. According to the Jean Ziegler UN special report on the right to food (For 2000 to March 2008), mortality due to malnutrition accounted for 58% of the total mortality in 2006 one out of twelve people worldwide is malnourished²⁷.

So, we planned to make the herbal product with the full of nutritional value. Hence, we selected the drug Cassia tora, Moringa oleifera, Asparagus racemosus which contain immense amount of carbohydrates, proteins, and fats minerals amino acid etc.

MATERIAL AND METHODS:

Collection and Authentication of crude drugs:

The seeds of Cassia tora was collected from the Manas Ayurveda store, Nagpur and subjected to shed drying, similarly the leaves of Moringa oleifera was collected from the Dadasaheb Balpande College of Pharmacy, Besa, Nagpur and dried indoor in dry place under the shady area. The dried roots of the Asparagus racemosus collected Manas Ayurveda store, Nagpur. and Authentication of herbs of Cassia tora, Moringa oleifera and Asparagus racemosus, was done by the Department of Botany, RTMNU, Nagpur.

Extraction process:

1) Extraction of seeds of Cassia tora:

Cassia tora seeds were crushed to a coarse powder and extracted with ethanol using a Soxhlet extractor. The procedure was repeated four times, and the ethanol was evaporated, yielding a semisolid alcoholic extract of Chakramarda and subjected to phytochemical screening²⁸.

2) Extraction of leaves of Moringa oleifera:

The maceration was done for dried powdered leaves of MO with Ethanol: water (1:1) at 45 C±2 for 48hours. After the maceration period it was filtered, concentrated to 10% of its original volume and dried in a hot air oven until a constant dry mass was obtained which was exposed to phytochemical screening²⁹.

3) Extraction of roots of Asparagus racemosus:

The dried roots of Asparagus racemosus were pulverized, defatted with hexane, and extracted using 90% methanol for 24hours. The methanolic extract was evaporated, producing a concentrated liquid syrupy mass dissolved in 10% methanol and phytochemical screening was conducted³⁰.

Phytochemical screening tests:

The qualitative phytochemical evaluation was done for the carbohydrates, proteins, amino acid, steroids, glycosides, tannins, phenols, alkaloids, flavonoids, vitamins A/C/D, and Minerals like iron, chloride, sulfate and magnesium with antioxidants³¹^,³².

Chromatographic analysis:

Thin-layer chromatography (TLC) was performed using different mobile phase to determine the R_f values of the plant extracts.

Antioxidant activity:

The plant extract's antioxidant capacity was assessed through the utilisation of the DPPH free radical Scavenging test³³. Using in vitro models, 1, 1-diphenyl-2-picrylhydrazyl radical techniques were used to measure the extract's antioxidant activity at various doses³⁴. 1 millilitre of the 0.3mM DPPH solution in 100% ethanol was created, and 3millilitres of the fraction dissolved in ethanol at different concentrations were combined with 1millilitre of this solution. A Shimadzu spectrophotometer was used to measure the absorbance of the mixture at 517nm after it had been stirred and allowed to stand at room temperature for 30 minutes. The extract was taken at various doses and let to stand at room temperature for 30 minutes. The absorbance was then measured using a UV Spectrophotometer, and the antioxidant activity (% inhibition) and IC₅₀ were calculated. The results were displayed on a graph³⁵^,³⁶.

Absorbance of control – Absorbance of Sample

% Inhibition activity = -------------------------------- × 100

Absorbance of Control

Nutritional profile:

1) Estimation of crude fibre:

True cellulose makes up the majority of crude fibre, with 97% of it being insoluble lignin and a small amount of mineral components. It only makes up 4%–6% of the lignin and 60%–80% of the cellulose³⁷. When moisture and fat free sample is boiled first with dilute acid (1.25 % H₂SO₄) and after with dilate alkali NaOH the soluble carbohydrate and protein transferred into the solution and are extracted³⁸. The undissolved residue which is left behind represents cellulose, hemicellulose and lignin combinely are called as crude fibres. When the remaining residue ether drying and weighing is heated at 60°C, the fibre content is obtained by removing the weight of ash from the weight of dried residue.

Procedure:

Accurately weigh 2-3g of the material and transfer it to a clean beaker. Add 1.25% H₂SO₄ and heat in a flask with a reflux condenser for 30 minutes. Filter the beaker's contents through a muslin cloth, washing the residue with boiling water until free from acid. Return the content to the beaker, add 1.25% NaOH (200 cc), and heat for 30 minutes. Filter again, washing the residue with boiling water until free from alkali. Transfer the residue to a dry silica container, dry and heat for 2-3 hours. Ignite the residue at 600°C in a muffle furnace for 30 minutes, then cool and weigh it.³⁹

Loss in wt after ignition in grams

% Crude fibre = ----------------------------------------- ×100

Wt of the original sample in grams

2) Estimation of Crude protein:

Nitrogen estimate is done using this process. This nitrogen value is used in a formula to determine crude protein. The amount of proteins in the feed or sample is measured indirectly by multiplying the nitrogen content by 6.25 to get the protein content40 after first calculating the nitrogen content⁴⁰.

Procedure:

Kjeldahl Method⁴¹^,⁴²

The Kjeldahl technique is the most conventional method of nitrogen content estimation. The approach consists of three main steps:

a) Digestion: Weigh about 2g of the drug sample into a Kjeldahl flask and add 25ml of concentrated sulphuric acid, 0.5g of copper sulphate, 5g of sodium sulphate, and a pinch of selenium pill. Heat the fume cupboard carefully to avoid excessive foaming, and continue to digest for roughly 45 minutes, or until the digest turns clear pale green in colour. Allow to cool fully before quickly adding 100ml of distilled water. Rinse the digesting flask twice or three times, then add it to the bulk⁴³.

b) Distillation: Distillation is carried out using a Markham distillation equipment. Heat the distillation equipment, then pour around 10ml of the digest through a funnel and bring it to a boil. Add 10ml of NaOH solution from the measuring cylinder to ensure that no ammonia is lost throughout the procedure. Distilled into 50mls of 2% boric acid, with screened methyl red as an indicator.

c) Titration: The alkaline ammonium borate produced is titrated against 0.1N hydrochloric acid, and the amount of acid utilised is noted⁴⁴.

3) Estimation of ether extract procedure:

To extract moisture from a drug sample, weigh 2-5 grams and transfer it to a filter paper or thimble. Plug the thimble with absorbent cotton. Insert the drug-filled thimble into the Soxhlet extractor, ensuring it remains below the bent of the extractor. Pour petroleum ether into the extractor using a glass funnel and assemble the Soxhlet apparatus. Plug the condenser tip with cotton and use approximately 1½ times the extractor's capacity of petroleum ether. Position the apparatus on a 40°C heater plate, securing it with clamps on a retort stand. Start circulating cold water through the condenser. After precisely 8 hours, remove the thimble with the material from the extractor. Reassemble the Soxhlet extractor and heat it on the heater plate to recover all remaining substances from the receiver flask, which will contain only crude fat and traces of petroleum ether. Remove the flask, wipe its exterior thoroughly to remove moisture and dust, then dry it in a hot air oven at 100°C for 60 minutes. Cool it in a desiccator, and finally, weigh it^45,46.

Wt of flask + extract – Tare wt of flask

% Ether Extract = ------------------------------------- × 100

Wt of the original sample in grams

4) Estimation of total ash value^47,48

Weight of ash

% Ash content = ---------------- × 100

Wt of sample

5) Estimation of Nitrogen free extract⁴²^,⁴⁹

The nitrogen-free extract is used to estimate non-fiberous carbohydrates like sugars and starches. The nitrogen free extract determination is utilized for proximate analysis and chemical analysis, followed by the necessary computations. NFE refers to soluble carbohydrates and other digestible and readily utilizable non-nitrogenous substances in drugs.

Nitrogen free extract % =100 % - [ % Crude fibre + % Crude protein + % Ether extract + % Total Ash]

Nutritive value is determined by⁵⁰

Nutritive value = 4 × Percentage of protein + 9 × Percentage of fat + 4 × Percentage carbohydrate

RESULT:

Practical yield:

Table 1: % practical yield of the product after extraction process

Sr. No.	Plant Extract	Practical yield (% w/w)
1	Cassia tora	8.3
2	Moringa oleifera	5.4
3	Asparagus racemosus	7.6

Ash value:

Table 2: Result of ash value determination

Sr. No.	Constants	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
1	Total ash value (%)	4.88	14.2	4.06
2	Water soluble ash value (%)	2.32	3.10	2.53
3	Acid insoluble ash value (%)	1.64	1.70	1.20

Phytochemical screening:

Table 3: Phytochemical analysis of Cassia tora, Moringa oleifera, Asparagus racemosus

Sr. No.	Chemical test for phytoconstituents	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
1	Carbohydrate	+	+	+
2	Proteins	+	+	+
3	Amino acids	-	-	-
4	Steroids	-	-	+
5	Glycoside	+	-	+
6	Tannins and phenols	+	+	+
7	Alkaloids	+	+	+
8	Flavonoids	+	+	+
9	Vitamin A Vitamin C Vitamin D	+ + -	+ + -	+ + -
10	Iron Chloride Sulphate Magnesium	- + + +	- + + +	- + + -
11	Antioxidants	+	+	+

Thin layer chromatography study:

Table 4: Thin layer chromatographic evaluation

Parameters	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
TLC Plates
Mobile phase	Chloroform: Methanol (18:2)	Toluene: 1-butanol: Methanol (9.5:0.5:0.5)	Ethyl acetate: Methanol: H₂O (7.5:1.5:1)
Rf values	0.75	0.75	0.61

Antioxidant Activity by DPPH scavenging assay:

Table 5: Result of % inhibition activity

Sr. No.	Concentration of extract (mg/ml)	Standard (Ascorbic acid)	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
1.	20	53.97	8.63	6.18	3.65
2.	40	62.15	19.00	10.43	15.18
3.	60	70.65	35.63	31.87	41.02

Absorbance of Control = 0.7392

Table 6: Comparison of IC₅₀ Values between standard and prepared herbal extract

Compounds	Standard (Ascorbic acid)	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
IC₅₀	13.92	82.83	92.68	72.16

Nutritional value:

Table 7:Result of Nutrients estimation

Sr. No.	Phytoconstituents	*Cassia tora*	*Moringa oleifera*	*Asparagus racemosus*
1	Crude fibre (%)	13.9	7.71	17.4
2	Crude Protein (%)	18.35	16.5	6.88
3	Ether extract (%)	5.54	2.5	2.14
4	Total ash (%)	4.88	14.2	4.06
5	Nitrogen free extract (%)	57.33	59.09	69.52

Nutritive Value:

Table 8: Nutritive Value of Cassia tora, Moringa oleifera, and Asparagus racemosus

Sr. No.	Plants	Nutritive value (Cal/100gm)
1	Cassia tora	178.86
2	Moringa oleifera	119.34
3	Asparagus racemosus	116.38

DISCUSSION:

The current study shows that Cassia tora, Moringa oleifera, Asparagus racemosus shows presence of carbohydrates, proteins, flavonoids, glycosides, alkaloids, tannins, terpenes, amino acids, vitamins, minerals and antioxidants. The main aim of the present research study is to estimate the nutritional value like crude fiber, protein, ether extract, total ash and nitrogen contents and antioxidant activity. The results shows that Cassia tora, Moringa oleifera, Asparagus racemosus contains a notable number of nutritional contents. The extract of these drugs subjected to the qualitative and quantitative evaluation of metabolite includes phytochemical evaluation, chromatographic evaluation, and nutritional evaluation with the values of their nutritional supplements and the drugs showed the satisfactory results.

CONCLUSION:

In conclusion the data derived from nutrient characterization of Cassia tora, Moringa oleifera and Asparagus racemosus clearly indicate that the plants are rich in nutrients and has potential to be used as a preservative with multiple purposes. They all are good nutrition as well as inexpensive and can be used to prevent a lot of disease. Nutritional supplements have the positive impact in case of malnutrition. Condition particularly herbal drug containing rich amount of carbohydrates, protein, amino acid, fats, minerals, tannins, alkaloids glycosides and antioxidants. So, from this study it can be concluded that the formulation prepared from the Cassia tora, Moringa oleifera, Asparagus racemosus, can show the positive effect on the malnutrition, so their formulation can use for the prevention and treatment of this malnutrition.

ACKNOWLEDGEMENTS:

The authors express their gratitude to Manas Ayurveda store, Nagpur for providing us with the seeds of Cassia tora and roots of Asparagus racemosus also thanks to Dadasaheb Balpande College of Pharmacy Besa, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur for constant support and encouragement.

DISCLOSURE STATEMENT:

No potential conflict of interest was reported by the authors.

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Received on 17.05.2023 Modified on 26.02.2024

Research J. Pharm. and Tech 2024; 17(10):4751-4757.

DOI: 10.52711/0974-360X.2024.00732