INTRODUCTION:

Ethnomedicinal plants, vegetables, fruits and herbs used in the folk and traditional medicine are currently considered as one of the main sources of cancer chemoprevention drug discovery and development. Over 60% of the clinically used anticancer drugs are of natural origin and most of them are derived from higher plants¹. Legume is a crop grown mainly for its edible seeds. It is widely grown in the tropics and subtropics, being important to both small and large commercial producers as well. Legumes are rich in nutrients, phytonutrients, niacin, protein, lipidsandfattyacids², folate, fiber, magnesium, vitamin E, manganeseandphosphorous³, stilbenephytoalexins⁴, stilbenoids^4,5, p-coumaric acid^6,7. Peanuts are also rich in many micronutrients and bioactive constituents⁸. It was also found to have many pharmacological effects such as antioxidant: hypolipidemic, anti-inflammatory, analgesia mediated by opioid receptor affinity, sympathomimetic, endocrine, antimicrobial, antiparasitic, sedative, hypotensive and haemostatic were attributed to the constituents of Arachis hypogaea.

High nutritional content is attributed to the presence of biologically active compounds such as, tocopherols, flavonoids, phytosterols, resveratrol, as well as to their relatively high level of protein and their easy oil digestibility⁹. More than 12 fatty acids have been reported in groundnut, among that oleic acid was reported to have beneficial effect to decrease the low-density lipoprotein levels, suppress tumorigenesis and ameliorate inflammatory diseases¹⁰. It has been reported that roasting promotes the antioxidant capacity of the intact peanuts due to the formation of the Millard reaction products¹¹. The conjugates formed by Maillard reaction generally exhibit improved features such as emulsifying properties, antioxidant activity, thermal stability, solubility and immuno-modulatory activities of polysaccharides¹². The high total phenolic content showed high antioxidant activity and can inhibit cancer cell growth effectively¹³. From the previous study it was analyzed that roasted Spanish groundnut variety exhibited high antioxidant property, hence the objective of the present study was to evaluate the effect of roasting on the concentrations of various Phyto-constituents, biomolecules, minerals, antioxidants and enzymes and compare with raw Spanish variety.

MATERIALS AND METHODS:

The two major variety of groundnut that is commonly found in Tamilnadu are Runner and Spanish and hence they were chosen for the study. The groundnuts were bought from the local market in Chennai and the seed were removed. These two varieties have contrast characteristics in terms of seed colour of the seed coat and size of the seed. The Runner variety groundnut are big sized seeds with pale pink colour, whereas the Spanish variety has dark red coloured small sized seeds. Roasted Spanish variety was found to be more effective from our previous study and hence the present study were done only with roasted Spanish (85 ºC for 20 min) and compared with raw Spanish. The seeds were then broken into coarse granules using mortar and pestle. 50gms were weighed and used for extraction process. The ethanolic extraction was carried out using Soxhlet extractor. The groundnut samples were packed in the thimble and was extracted using ethanol, the extraction process was carried out at 74 ^oC for 4 hours. These extracts were then used for the estimating Phytochemical such as flavonoids¹⁴, alkaloids¹⁵, phenols¹⁶, tannins¹⁷, saponins¹⁸ and biomolecules such as total protein by the method of Lowry etal., (1951) and albumin by BCG method.^19,20, Total cholesterol, triglycerides and HDL- cholesterol by enzymatic methods^21,22,23, Calcium and magnesium were diagnosed by (OCPC) method- Gitelman (1967) and Calmagite, Gindler et al method (1971)^24,25, Potassium²⁶, chloride by using mercuric chloride and diphenylcarbazone²⁷, Glucose by Glucose oxidase method²⁸, vitamin E and vitamin C by Desai (1984) and Omaya et al (1979)^29,30, and Iron³¹. Six albino rats were used for the analysis, two as a control and four as a test model. Institutional ethical clearance was obtained to carry out the study. The test animals were fed with the 5g of groundnut per day at four intervals and the control were fed with normal diet. The animal was sacrificed on the third and fifth day; the liver was removed and homogenized using mortar and pestle. The homogenized liver was then centrifuged and the supernatant were used for invivo analysis of liver marker Enzymes like SGOT³², SGPT³³ and Antioxidants such as the level of lipidperoxidation (TBARS) by the method of Nichans and Samuelsson(1968)³⁴. Glutathione was determined by Ellman (1959)³⁵, vitamin C²⁹, vitamin E³⁰ and Biochemical parameters such as Carbohydrates ²⁸, protein¹⁶, lipids^21,22,23, minerals^24,25,26, bilurubin³³, excretory product like Urea by Natelson et al., (1951) , uric acid by Caraway, (1963) , creatinine by the method of Owen et al., (1954)^36,37,38. The data for biochemical analysis were expressed as mean± S.D. Obtained data were analyzed by using independent sample t test by Statistical Packages for Social Science Software (SPSS10.0). Statistical analysis with the value of P < 0.05 is considered as significant.

RESULTS AND DISCUSSION:

Biochemical analysis was carried out to find the concentration of different phytochemicals and biomolecules in both roasted and non-roasted peanuts of Spanish variety, as the evidence created by the previous study that Spanish variety exhibited high antioxidant property.

TABLE 1: Phytotochemical Concentrations in Raw and Roasted Extracts of Spanish Arachis hypogaea.

S. No	Parameter	Non-roasted (mg/g)	Roasted (mg/g)
1	Alkaloid	6000 mg/g	6600 mg/g
2	Flavanoid	260 mg/g	320 mg/g
3	Phenol	218 mg/g	380 mg/g
4	Tannins	903mg/g	1205 mg/g
5	Saponin	480mg/g	489 mg/g

TABLE 2: Concentration of Biomolecules in Raw and Roasted Extracts of Spanish Arachis hypogaea

S. No	Parameter	Non-roasted (mg/g)	Roasted (mg/g)
1	Protein	2.75 g/lit	0.63 g/lit
2	Cholesterol	4.45 g/lit	5.5 g/lit
3	Calcium	0.043 g/lit	0.2816 g/lit
4	Potassium	0.693g/lit	0.996 g/lit
5	Chloride	0.866 g/lit	3.885 g/lit
6	Glucose	4.3 g/lit	2.64 g/lit
7	Magnesium	3.1 g/lit	9.388 g/lit
8	Vitamin C	0.733 g/lit	0.133 g/lit
9	Vitamin E	0.0815g/lit	1.05 g/lit
10	Iron	0.98 g/lit	1.8 g/lit

The consumption of a plant-based or phytochemical rich diet has been associated with a reduced risk of chronic human illnesses such as certain types of cancers, inflammation, cardiovascular and neurodegenerative diseases³⁹. Peanuts (Arachis hypogaea.) have a remarkable nutrient profile as well as bioactive components and hence exhibit different pharmacological activities. In the present study the levels of phytoconstituients such as Saponins, flavonoids, Alkaloids, phenols and Tannins were found to be higher in roasted Spanish groundnut when compared to the raw groundnut. Saponin inhibit microbial proliferation and also have hypolipidemic and anticancer activity. The saponin reacts with cholesterol rich plasma membrane of various cancer cells and arrests their proliferation⁴⁰. Flavonoids possess anti-tumoral, hypersensitive, and anti-inflammatory activities^41,42,43. Alkaloids have been reported to be effective against Human Immunodeficiency virus (HIV) infection⁴⁴. The highest value of tannins was in A. hypogaea seeds having astringent properties for healing of wounds and inflamed mucous membrane⁴⁵. The antioxidant properties of plant extracts are attributed to their polyphenolic contents⁴⁶. Hence this study reveals that the pharmacological activity of roasted groundnut was more effective compared to non-roasted groundnut. Table 2: depicts the nutritional contribution of roasted peanuts in comparison to the raw groundnut. The level of protein, vitamin C were reduced on roasting whereas the levels of cholesterol, glucose calcium, potassium, chloride, magnesium, iron and vitamin E were found to be increased on roasting. The level of protein for roasted peanut is lower than raw peanut could be attributed to denaturation as reported previously⁴⁷. The increase in the concentration of biomolecules in the roasted extract might be due to Maillard reaction, where they serve as the precursors of Millard compounds.

In Vivo Analysis after Third Day Treatment:

Table-3: Lipidperoxidation and Antioxidant status in Rat Liver tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	LPO (millimole/ml)	8.995±0.76	64.79±0.92	8.19±0.76 *
2	GSH (µg/dl)	364±.13.0	352±8.0	121±12.5*
3	VIT-C (mg/dl)	4.15±0.12	10.13±0.10	4.21±0.07*
4	VIT-E (mg/dl)	0.13±.008	0.171±0.007	0.18±0.006*

Values are expressed as mean ± SD. *P< 0.05 is significant

Table-4: Effect of Arachis hypogaea on Biomolecules in Rat Liver Tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1.	Glucose(mg/dl)	80.8±1.0	91±1.09	81.85±1.33*
2.	Protein(g/dl)	7.0±.64	8.05±0.05	6.1±0.08
3.	Albumin(g/dl)	2.9±0.10	3.86±0.33	2.75±0.26
4.	Cholesterol((mg/dl)	228.8±20.4	210.8±10.2	145.6±3.38*
5.	TG (mg/dl)	95.43±3.36	74.5±1.04	70.86± 3.2*
6.	HDL (mg/dl)	18.7±0.54	23.23±1.92	31.6±0.51*
7.	Calcium	6.25±0.41	10.33±0.51	11.9±0.49*
8.	Potassium	111.93±0.51	172.5±1.22	113.33±3.26*
9.	Chloride	277.16±1.72	386.16±3.81	255.16±3.31
10.	Iron	215.6±1.36	251.33±1.36	164.2±1.09

Values are expressed as mean± SD. *P< 0.05 is significant

Table-5: Liver marker enzymes activity in Rat Liver tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	SGOT(IU/L)	48.2±0.68	75.33±1.03	46.16±0.98*
2	SGPT(IU/L)	54.8±0.98	72.5±0.54	61.66±0.81*

Values are expressed as mean ±SD. *P< 0.05 is significant

Table-6: Bilirubin and Excretory product concentration in Rat Liver tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	Total bilirubin(mg/dl)	1.06±0.05	1.866±0.05	1.35±0.13*
2	Direct bilirubin(mg/dl)	1.20±0.041	1.48±0.04	1.26±0.05*
3.	Urea(mg/dl)	34.5±0.54	62.66±1.03	47.3±0.81*
4.	Uric acid(mg/dl)	4.28±0.22	9.33±0.31	5.15±0.13*
5.	Creatinine(mg/dl)	1.08±0.98	1.36±0.05	1.13±0.03*

Values are expressed as mean± SD. *P< 0.05 is significant

After Fifth Day Treatment:

Table-7: Lipidperoxidation and Antioxidants

S. No	Parameter	Normal	Nonroasted	Roasted
1	LPO (millimole/ml)	9.295±0.36	68.79±0.32	10.12±0.76*
2	GSH(µg/dl)	464±.13.0	403±12.5	148±8.0*
3	VIT-C(mg/dl)	5.12±0.12	12.13±0.10	5.21±0.07*
4	VIT-E(mg/dl)	0.18±.008	0.189±0.007	0.26±0.006*

Values are expressed as mean± SD. *P< 0.05 is significant

TABLE-8: Effect of Arachis hypogaea on Biomolecules in Rat Liver Tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	Glucose(mg/dl)	80.66±1.03	82.5±2.7	101.6±1.86*
2	Protein(g/dl)	7.05±0.32	7.6±0.18	8.41±0.14
3	Albumin(g/dl)	4.68±0.13	3.65±0.12	4.13±0.17
4.	Cholesterol((mg/dl)	152.5±7.23	96.66±1.5	93.5±2.25*
5.	TG (mg/dl)	46.33±0.51	30.53±0.196	28.46±0.51*
6.	HDL (mg/dl)	20.6±0.098	33.75±1.89	34.75±1.62*
7.	Calcium(mg/dl)	10.58±0.38	12.22±0.30	14.4±0.38*
8.	Potassium(mg/dl)	113.83±1.47	145.2±1.32	148.2±1.40*
9.	Chloride(millimole/l)	82.66±1.86	127.16±0.752	107.33±0.81
10.	Iron(µg/dl)	205.6±1.24	221.33±1.20	144.2±1.06

Values are expressed as mean± SD. *P< 0.05 is significant

Table-9: Liver Marker Enzymes activity in Rat Liver Tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	SGOT(IU/L)	45.8±0.58	78.2±0.51	49.2±0.92*
2	SGPT(IU/L)	52.1±0.85	70.5±0.56	59.8±0.86*

Values are expressed as mean± SD. *P< 0.05 is significant

TABLE-10: Bilirubin and Excretory Product in Rat Liver Tissue homogenate

S. No	Parameter	Normal	Nonroasted	Roasted
1	Total bilirubin(mg/dl)	1.09±0.06	1.92±0.06	1.62±0.12*
2	Direct bilirubin(mg/dl)	1.2±0.040	1.58±0.03	1.15±0.32*
3.	Urea(mg/dl)	34.1±0.43	60.6±1.02	46.2±0.72*
4.	Uric acid(mg/dl)	4.6±1.02	8.98±0.42	5.3±0.12*
5.	Creatinine(mg/dl)	1.06±0.62	1.42±0.04	1.16±0.08*

Values are expressed as mean± SD. *P< 0.05 is significant

Table 3 and 7 represents In vivo lipid peroxidation and antioxidant status in the 3^rd and 5^th day in experimental animal and compared with control. The level of LPO and GSH was found to decrease significantly in roasted groundnut fed rat compared to raw and control animal, whereas Vit C and Vit E were found to increase significantly in roasted groundnut fed rat than the other group. The energy store in peanut is in the form of unsaturated fatty acid and this might have prevented lipid oxidation. This observation is consistent with the previous report that roasting enhance antioxidant effect thereby degrade natural toxins and enzymes⁴⁸. The decrease in GSH might be due to increased utilization to trap hydro peroxides. The increase in Vit C and Vit E is in accordance with Abudu et al., 2004⁴⁸. Table 4 and 8 represents the biomolecules level in 3^rd and 5^th day peanut fed animals. In our present study the level of Glucose, Protein, Albumin, Cholesterol, HDL, T.G, Calcium, Potassium, were significantly higher in roasted ground fed rat compared to the other groups, the level were found to increase as the days increases. But the level of Iron and chloride were found to be lower in roasted peanut fed rat when compared to raw and control. Sucrose is abundantly present in peanut, hence sucrose get hydrolyzed on roasting thereby glucose concentration is increased⁴⁹. In our present study the decrease in Total Cholesterol, T.G and increase in HDL in roasted groundnut compared to raw and control might be due to increase amount of unsaturated fats (MUFAs and PUFAs)⁵⁰. Roasting of peanut result in decrease solubility⁵¹ hence the total protein content and Albumin was found to decrease in roasted extract fed animal compared to non- roasted. Roasting also increase the level of Calcium and potassium, the level of chloride and Iron were found to decrease significantly compared to raw peanut fed animal. The level of SGOT, SGPT and bilirubin level were found to decrease in roasted groundnut fed animal indicating hepato protective activity. In the present study, the levels of urea uric acid and creatinine were higher in raw peanut treated group in comparison to those of control group⁵² whereas the level was found to decrease in roasted peanut fed animal. Peanuts significantly reduce renal oxidative damage and increase nitric oxide levels. Thus, suppress toxin mediated enhancement of serum urea and creatinine levels⁵³ some other researchers suggested that the peanut keeps the serum creatinine level close to normal due to the ability of some antioxidant in peanut to scavenge free radicals generated by irradiation, which would otherwise cause kidney damage⁵⁴. This study clearly reveals that roasting improves the nutritive value and the antioxidant status of the groundnut. Hence it is beneficial to consume roasted groundnut instead of raw groundnut.

CONFLICT OF INTERESTS:

The authors declares that there is no conflict of interests regarding the publication of this paper.

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Received on 02.08.2018 Modified on 19.09.2018

Research J. Pharm. and Tech. 2019; 12(3): 1239-1243.

DOI: 10.5958/0974-360X.2019.00206.3