EFFECT OF SOLVENTS ON ANTIOXIDANT ACTIVITIES OF FERONIA LIMONIA FRUIT

EFFECT OF SOLVENTS ON ANTIOXIDANT ACTIVITIES OF FERONIA LIMONIA FRUIT

R. Srivastava ¹, N. Mishra ², S. Tripathi ¹ and N. Mishra ^{* 3}

Centre of Food Technology ¹, Department of Home Science ³, University of Allahabad, Prayagraj - 211002, Uttar Pradesh, India.

Ethelind School of Home Science ², Sam Higginbottom University of Agriculture, Technology and Sciences, Prayagraj - 211010, Uttar Pradesh, India.

ABSTRACT: Feronia limonia (F. limonia) is an underutilized edible fruit commonly known as kaitha. The study was performed to compare the effect of three solvents at different concentrations on antioxidant activities of F. limonia fruit. Fruits were extracted in different solvents viz., ethanol, methanol, and acetone at concentrations of 50%, 70%, and 90%. Each extract was analyzed for total phenol and flavonoid content and their antioxidant activities such as Free radical scavenging activity (DPPH), Ferric reducing antioxidant power (FRAP), Metal chelating activity and reducing capacity (RC). The results indicate that 70% aqueous acetone works best for the extraction of TPC (35.60 mg GAE/g), TFC (31.69 mg QCE/g), and other antioxidant activities. The phenol and flavonoid content showed a significant correlation with antioxidant activities. The findings of the study suggested that F. limonia fruit is a good and cheap source of natural antioxidant which can serve as a functional food. The fruit possesses potent enormous health benefits and thus may be used in food and pharmaceutical applications.

Feronia limonia, Solvents, Total phenol content, Total flavonoid content, Antioxidant, bioactive compounds

INTRODUCTION: Feronia limonia (F. limonia.) plant, which belongs to the Rutaceae family is a native plant of India. It grows in India, Pakistan, China, and other Southeast Asian countries. The shape of the fruit is spherical, with 5-12.5 cm in diameter. The rind of the fruit is hard, woody and very difficult to crack ¹. In India, the fruit is consumed as syrup, drink, murabba, chutney, and juices. Fruit posses enough therapeutic potentials including anti-hypercholesterolemic, antidiabetic, anticarcinogenic, antimutagenic, anti-inflammatory and antibacterial activities due to the presence of bioactive and antioxidant compounds.

The antioxidant properties of F. limonia fruit could be attributed to phenolic compounds, phenolic gly-cosides, flavonoids, tannins, coumarin, terpenoid, marmesin and luteolin ^2-3. These compounds help to decrease the level of Glutathione (GSH), Glutathione peroxidase (GPX), Superoxide dismutase (SOD) and Catalase (CAT). Therefore it is used in the treatment of various diseases such as diarrhea, dysentery, chest pain, kidney stones, painful cramps, and ulcer ⁴.

Antioxidant activity of many plants and their therapeutic effects were studied in recent years. This is frequently involved in various mechanisms such as inhibition of free radical generation as well as enhancement of scavenging capacity against free radicals and so on. Antioxidants were studied to develop natural antioxidant-rich food, cosmetics, and other products ⁵. The regular consumption of natural antioxidants has long been associated with the prevention of several diseases such as diabetes, cardiovascular diseases and cancer ⁶. Therefore, finding new and safe antioxidants from natural sources is of great interest now a day. Since biologically active compounds occur naturally in very small concentrations and presence of various antioxidant compounds with different chemical characteristics may or may not be soluble in a particular solvent; therefore, the choice of a suitable concentration of solvent is an important step in antioxidant studies ⁷. The polarity of the solvents plays an important role in the extraction of bioactive compounds. Several previous studies reveal that mixture of solvent with water is an effective method of extraction of bioactive compounds from natural sources.

The compounds such as phenols and flavonoids showed a preventive effect against degenerative diseases such as diabetes, cardiovascular disease, cancer, obesity, and inflammations. Phenol shows their redox abilities, which are a class of antioxidant agents that can quench and neutralize the free radicals ⁸. The objective of this work was to investigate the effect of three solvents at different concentrations on the extraction of phenols and flavonoids content of F. limonia fruit and to their antioxidant activity of the extracts by in-vitro method. The study also investigated the correlation of TPC and TFC with antioxidant activities including DPPH free radical scavenging activity, FRAP, metal chelating activity and reducing capacity.

MATERIALS AND METHODS: Unripe F. limonia fruit used in this study was purchased from the local market of Prayagraj (Allahabad), India. All the chemical used for the study includes foline ciocalteu reagent, methanol, ethanol and acetone, gallic acid, sodium nitrite, sodium hydroxide, trichloroacetic acid, ferric chloride anhydrous, ascorbic acid, sulphuric acid, and potassium-hexacyanoferrate, anhydrous monobasic potassium phosphate, sodium carbonate anhydrous, 2,2- diphenyl-1-picrylhydrazyl (DPPH), and aluminum chloride anhydrous,  were purchased from Sigma-Aldrich GmbH (Sternheim, Germany), Merck (Darmstadt, Germany) and Sigma-Aldrich (St. Louis, MO, USA).

Sample Preparation: Fruits of F. limonia were washed with water, and their rinds were removed. Fresh F. limonia pulp was oven-dried at 40 ºC. The dried samples were ground into powder and passed through a 60-mesh sieve. Then 1 g powder was mixed in 100 ml solvents at the different concentrations and kept a room temperature for 24 h. The solid-liquid mixture was obtained which was centrifuged using centrifuged for 10 min at 1000 rpm. The supernatant of the solid-liquid mixture was collected within the amber-colored glass bottle and the extracts were stored in the refrigerator at 4 ºC for further analysis.

Determination of Total Phenol Content (TPC): Folin-ciocalteu method was used to determine the TPC content of sample ⁹. 0.2 ml of each extract was taken in test tubes and added 5 ml of 10% diluted folin-ciocalteu phenol reagent. 4 ml of 7.5% sodium carbonate solution was added within 5-10 min. The mixture is allowed to stand for 60 min in dark. TPC was determined using spectro-photometric method (Model Evolution 600, Thermoscientific, US) by comparison of standard plots at 765 nm. A standard curve was prepared using gallic acid solutions (20, 40, 60, 80, 100 mg/L). The absorbencies were recorded at 765 nm. The results were expressed as mg of gallic acid equivalents / g of samples on a dry basis.

Determination of Total Flavonoid Content (TFC): Aluminum chloride colorimetric assay was used to measure the TFC content of the sample. 2 ml of sample extracts were taken in test tubes added 150 µl of 5% NaNO₂ shake the mixture and left it for 5 min, added 150 µl of 10% AlCl₃ in the test tubes_. After 10 min, 1 ml of 1 mol sodium hydroxide (NaOH) was added and 10 ml total volume was made with the help of distilled water. The mixture was vortex and after keeping the mixture in incubation for 10 min and the absorbance was recorded against a blank at 510 nm¹⁰. A calibration curve was prepared using a standard solution of quercetin (20, 40, 60, 80 and 100 mg/L). The results were expressed as mg quercetin equivalent (QCE)/g of sample on a dry basis.

Antioxidant Activity:

DPPH Free Radical Scavenging Activity: DPPH free radical scavenging activity was evaluated by 1, 1-diphenyl 2-picryl-hydrazil (DPPH). 100 µl of fruit extract was mixed with 150 µl of 0.1 mmol DPPH methanol solution and incubated for 15 min. Absorbance was measured using the spectrophoto-meter (Model Evolution 600, Thermoscientific, US) at 515 nm ¹¹.

DPPH activity was calculated using the formula:

(A control – A sample / A control) × 100

Where A control denotes the absorption of the DPPH solution and A sample, which is the absorption of the DPPH solution after the addition of the sample. Results were expressed as % of inhibition of the DPPH radical.

Ferric-Reducing Antioxidant Power (FRAP) Assay:  Sodium acetate buffer (300 mmol, pH 3.6), 10 mmol TPTZ solution (40 mmol HCl as solvent) and 20 mmol iron (III) chloride solution in a volume ratio of 10:1:1 respectively were used for preparing FRAP reagent. 200 µl extract of the three solvents at different concentrations was taken in test tubes and 1.3 ml of FRAP reagent added. The mixture was incubated for 30 min at 37 ºC. Absorbance was measured using spectrophotometer (Model Evolution 600, Thermoscientific, US) at 593 nm. The standard curve was prepared using FeSO₄.7H₂O solution (200, 400, 600, 800, 1000 μmol), and the results were expressed as μmol of ferrous equivalent Fe (II)/g of sample on the dry basis ^12-13.

Metal Chelating Activity: An aliquot (0.5 ml) of fruit extracts was mixed with 50 µl of ferrous sulfate. Add 1.6 ml of 80% ethanol and 100 µl ferrozine after 5 min. The mixture was vortexes for 1 min. After 10 min, absorbance was measured at 562 nm ¹⁴. The metal chelating activity was calculated using the ratio:

(1-absorption of sample /absorption of control) × 100

Results were expressed as % of inhibition of the ferrous sulfate of the sample on a dry basis.

Reducing Capacity (RC): 1 ml of different concentrations of extracts was mixed with 2.5 ml sodium phosphate buffer (pH 6.6) and 2.5 ml potassium ferricyanide (1% w/v). The mixtures were incubated for 20 min at 50 ºC. After that, 2.5 ml of TCA (10% v/v) was added and the samples were centrifuged at 10,000 rpm by 10 minutes. 2.5 ml of supernatant was mixed with 2.5 ml of distilled water and 0.5 ml of ferric chloride (0.1% v/v). The absorbance was subsequently measured at 700 nm with spectrophotometer¹⁵. The reducing power was related to ascorbic acid solution and expressed as µmol of ascorbic acid equivalents (AAE)/ g of dry weight.

Statistical Analysis: The analysis was carried out in triplicate and values were expressed as mean ± standard deviation. SPSS version 16.0 for Windows software (IBM corp.) was used for data analysis. Analysis of variance and Duncan’s multiple range method were used to compare any significant difference between the solvents. Differences were considered significant at P<0.05 and P< 0.01. All the figures are prepared using Microsoft excel. Correlations analysis between the antioxidant activities of the four independent tests (DPPH, FRAP, Metal Chelating Activity, and Reducing Capacity) with TPC and TFC were conducted using statistical package for social sciences (SPSS) 16.0.

RESULTS AND DISCUSSION:

TABLE 1: EFFECT OF SOLVENTS AT DIFFERENT CONCENTRATIONS ON TOTAL PHENOL AND FLAVONOID CONTENT OF F. LIMONIA FRUIT

GAE =Gallic acid equivalent; QCE =Quercetin equivalent, TFC =Total Flavonoid Content; TPC = Total Phenol Content, Analysis of variance P<0.05. Means carrying the same letter in superscript in a column do not differ significantly (P<0.05)

Total Phenol and Total Flavonoid Content: Table 1 shows the TPC value of the three solvents at different concentrations, ranging from 2.20 to 35.60 mg GAE/g. TPC value is decreasing in the following order: 70% aqueous acetone > 90% aqueous methanol > 70% aqueous methanol > 70% aqueous ethanol > 90% aqueous ethanol > 50% aqueous methanol > 50% aqueous ethanol > 50% aqueous acetone > 90% aqueous acetone. The highest TPC content shown by 70% aqueous acetone (35.60 mg GAE/g) and it is significantly different (P<0.05) from other concentrations while 90% aqueous methanol (28.57 mg GAE/g) and 70% aqueous methanol (25.46 mg GAE/g) used as second and third best solvent for TPC extraction respectively. The results of the present study were similar to previous reported studies ^{16, 17, 18}. Table 1 shows Total Flavonoid Content (TFC) of F. limonia fruit. The TFC of different solvents at varied concentration ranges from 4.56 to 31.69 mg QCE/ g. TFC is decreasing in the following order: 70% aqueous acetone > 50% aqueous acetone > 90% aqueous methanol > 70% aqueous ethanol > 70% aqueous methanol > 90% aqueous ethanol > 50% aqueous ethanol > 50% aqueous methanol > 90% aqueous acetone. The highest TFC content shown by 70% aqueous acetone (31.69 mg QCE/g) and the value is significantly different (P<0.05) from other concentrations while 90% aqueous acetone (4.56 mg QCE/g) shows the least TFC content.

Antioxidant Activities of F. limonia Fruit:

DPPH Free Radical Scavenging Activity: Fig. 1 shows the DPPH free radical scavenging activities of three solvents at different concentrations. The DPPH free radical scavenging activity by various solvents decreased in the following order: 70% aqueous acetone > 90% aqueous methanol > 70% aqueous methanol > 70% aqueous ethanol > 90% aqueous ethanol > 50% aqueous methanol > 50% aqueous ethanol > 50% aqueous acetone and 90% aqueous acetone. The results show that values obtained from the various polarity solvents were significantly different (P<0.05), and 70% aqueous acetone shows the highest (55.63%) value for DPPH free radical activity. The range of the DPPH free radical scavenging activity is varied from 10.63 to 55.63%. This indicates that 70% aqueous acetone extract is the most suitable solvent among all the three solvents at different concentrations for DPPH free radical activity of F. limonia fruit. The results indicate that 70% aqueous acetone extract shows the highest DPPH free radical scavenging activity (55.63%) and it is significantly different (p<0.05) from other concentrations of solvents. The extraction pattern of DPPH free radical scavenging activity in different concentrations of solvents followed the TPC extraction pattern.  The present study shows that both TPC and DPPH are following the same pattern of decrease in respective solvents. This could be due to high quenching and neutralizing power of phenol against free radicals ⁸. A similar trend was also observed in the study of DPPH radical scavenging activity of pineapple crude extract ¹⁹.

Ferric Reducing Antioxidant Power (FRAP): Fig. 2 shows that the FRAP value of  various solvents decreased in following order: 70% aqueous acetone > 50% aqueous acetone > 90% aqueous methanol > 70%  aqueous ethanol > 70% aqueous methanol > 90% aqueous ethanol > 50% aqueous ethanol > 50% aqueous methanol and 90% aqueous acetone. This study reveals both TFC and FRAP are following the same pattern of decrease in respective solvents. Findings of the FRAP analysis are shown that values obtained from various polarity solvents were significantly different (P<0.05) and 70% aqueous acetone reveals the highest (15.57 mmol of Fe (II) E /g) FRAP value suggested as the most suitable solvent among all the three solvents at different concentrations.

Metal Chelating Activity: In Fig. 3, among the three solvents at different concentrations, 70% aqueous acetone shows the highest metal chelating activity by various solvents decreased in the following order: 70% aqueous acetone > 50% aqueous acetone > 90% aqueous methanol > 70% aqueous ethanol >  70% aqueous methanol > 90% aqueous ethanol > 50% aqueous ethanol > 50% aqueous methanol and 90% aqueous acetone. It is significantly different (P<0.05) from other concentrations. Metal chelating activity for 70% aqueous acetone extract was the highest (94.54%). Both TFC and metal chelating is following the same pattern of decreasing in respective solvents. This indicates that the 70% aqueous acetone extract is the most suitable solvent among all the three solvents at different concentrations for the metal chelating activity of F. limonia fruit.

Reducing Power: In Fig. 4, all extracts of different solvents at different concentrations show some degrees of electron-donating capacity in a concentration-dependent manner. The highest reducing capacity of F. limonia fruit is shown by 70% aqueous acetone extract and is significantly higher (p<0.05) than that of the other extracts at all concentrations studied, followed by that of the 50% aqueous acetone, 90% aqueous methanol, 70% aqueous ethanol, 70% aqueous methanol, 90% aqueous ethanol, 50% aqueous ethanol, 50% aqueous methanol and 90% aqueous acetone. The present study shown both TFC and reducing capacity is following the same extraction pattern of decrease in respective solvents. The lowest reducing power was found in the 90% aqueous acetone extract. The value is also significantly lower than that of the other extracts at all concentrations studied. The analysis of results indicated that the reducing capacity of the 70% aqueous acetone was the highest (60.56 µmol AAE/g) and is significantly different (P<0.05) from other solvents.

Correlation between the Total Phenol, Flavonoid Content and Antioxidant Activities of F. limonia Fruit: Pearson’s correlation coefficient was applied to determine the relationship between the antioxidant activities include DPPH, FRAP, metal chelating activity and reducing capacity and TPC, TFC shown in Table 2.

TABLE 2: PEARSON’S CORRELATION COEFFICIENTS OF DPPH, FRAP, METAL CHELATING ACTIVITY AND REDUCING CAPACITY VERSUS TPC AND TFC

*Correlation is significant at ? < 0.01.

The TPC showed a strong correlation to DPPH with Pearson’s correlation coefficient of 0.88 and also revealed correlation to metal chelating activity and reducing capacity with Pearson’s correlation coefficient of 0.40 and 0.32 respectively while TPC shows a negligible correlation to FRAP with Pearson’s correlation coefficient of 0.25. A very strong correlation between the TFC versus FRAP, metal chelating activity, and reducing capacity was seen with Pearson’s correlation coefficient of 0.96, 0.94 and 0.98 respectively. The TFC shows a negligible correlation to DPPH with Pearson’s correlation of 0.24. The correlation coefficient between TFC versus FRAP, metal chelating activity, and reducing capacity were statistically higher than those between TPC versus FRAP, metal chelating activity and reducing capacity. Flavonoid seemed to have a higher correlation with various antioxidant activities than phenols. Several previous studies demonstrated that both phenols and flavonoids which are major antioxidants found in natural products possess antioxidant activity^20-22.

The present study confirmed that the antioxidant activities of selected fruit are attributed to both flavonoids and phenols. TFC shows the high influence on antioxidant activities as it is strongly correlated with FRAP, metal chelating activity, and reducing capacity with Pearson’s correlation coefficient of 0.96, 0.94, and 0.98, respectively while TPC shows a good correlation with Pearson’s correlation coefficient of 0.88. TPC was strongly correlated with DPPH free radical scavenging activity. These results may be explained by the interaction of chemical structural differences between phenol compounds and concentrations of the solvents used. TPC was highly correlated with DPPH in comparison to FRAP, metal chelating activity and reducing capacity while TFC was highly correlated with FRAP, metal chelating activity and reducing capacity. Hanchinalmath et al., ⁵ suggested that luteolin is a primary active ingredient of F. limonia fruit, which is a flavonoid in nature ^23-24. Thus, this specific flavonoid compound may effectively contribute to the antioxidant activities of F. limonia fruit. The findings of the present study reveal the particular flavonoid and phenolic compounds that correlated with the antioxidant activities of F. limonia fruit need to be further investigated. Our study supports to Jin Gan et al. ²⁵

CONCLUSION: It can be concluded that 70% aqueous acetone is the best solvents for the extraction of TPC, TFC, and antioxidant activity of F. limonia fruit. The correlation results and statically analysis of this study is suggested that the antioxidant activity of F. limonia fruit is not only because of phenols but also flavonoids. Thus, flavonoids and phenols both are the major substances that contribute to the antioxidant activity of F. limonia fruit. Flavonoids exposed to have a stronger correlation with antioxidants than phenols. There is a need to conduct more studies to identify flavonoid and phenolic compounds that are correlated with the antioxidant activity of F. limonia fruit. The results of the present study would provide a new approach for further studies to the mechanisms which attribute the special bioactive of F. limonia fruit.

ACKNOWLEDGEMENT: Rashmi Srivastava acknowledges the Junior Research Fellowship (JRF) from University Grants Commission (UGC), New Delhi, India.

CONFLICTS OF INTEREST: The authors declared no conflict of interest.

REFERENCES:

1. Jayashree VH and Londonkar R: In-vitro antioxidant activities of Feronia limonia Indo American Journal of Pharmaceutical Research 2014; 4(6): 2937-42
2. Reddy DK, Sushmitha G and Singh JK: Development and standardization of wood Apple-Dates Jam. Advanced Engineering and Applied Sciences: An International Journal 2016; 6(2): 69-73
3. Banupriya L and Poongodi VT: Anti nutrient and phytochemical screening of an underutilized fruit seed: Limonia acidissima. International Journal of Innovative Research in Technology 2016; 2(9): 7-14
4. Ponnuraj S, Jaganathan D, Kanagarajan M, Castro J and Darsini DTP: Influence of Limonia acidissima against the biofilm forming Aeromonas Hydrophila isolated from fresh water fishes. J Biochem Tech 2015; 6(1): 910-21.
5. Hanchinalmath JV and Londonkar R: Isolation and identification of a Flavone from fruit pulp of Feronia limonia. International Journal of Current Pharmaceutical Research 2014; 6(4): 28-31
6. Hanchinalmath JV and Londonkar R: Cytotoxic and apoptosis-inducing effect of luteolin isolated from Feronia limonia on hepg 2 cells. Biolife 2014; 2(4): 2320-57
7. Patel A, Shah D, Desai TR and Noolvi MN: Mucoadhesive buccal films based on chitosan and carboxymethylated Feronia limonia Fruit pulp mucilage interpolymer complex for delivery of opioid analgesics. Asian Journal of Pharmaceutics 2016; 10(2): 137-43.
8. Pandey S, Satpathy G and Gupta RK: Evaluation of nutritional, phytochemical, antioxidant and antibacterial activity of exotic fruit “Limonia acidissima”. Journal of Pharmacognosy and Phytochemistry 2014; 3(2): 81-88
9. Jayashree VH and Londonkar R: Comparative phytochemical studies and antimicrobial potential of fruit extracts of Feronia limonia International Journal of Pharmacy and Pharmaceutical Sci 2014; 6(1): 731-34.
10. Sonawane SK and Arya SS: Effect of drying and storage on bioactive components of Jambhul and wood apple. J Food Sci Technol 2015; 52(5): 2833-41.
11. Bhatt DK and Jha A: A study of incorporation of therapeutic values of wood apple (Feronia limonia Swingle) In Fruit Bar. Int J Pharm Sci Res 2015; 6(10): 4398-05.
12. Rakhunde PB, Saher S and Ali SA: Neuroprotective effect of Feronia limonia on ischemia reperfusion induced brain injury in rats. Indian Journal of Pharmacology 2014; 46(6): 617-21.
13. IIaiyaraja N, Likhith KR, Babu GRS and Khanum F: Optimisation of extraction of bioactive compounds from Feronia limonia (wood apple) fruit using response surface methodology (RSM). Food Chemistry 2015; 173: 348-54.
14. Tham NTH and Minh NP: Effect of ripen mature and ratio of wood apple’s pulp supplemented with sugar for Feronia limonia. International Journal of Advances in Pharmacy, Biology and Chemistry 2014; 3(2): 310-18.
15. Patel AS and Pandey AK: Fortification of Limonia acidissima Fruit powder to develop the phynolic enriched herbal biscuits. Journal of Bioresource Engineering and Technology 2014; 1: 74-85.
16. Bellah SF, Raju MIH, Billah SM, Rahman SE, Murshid GMM and Rahman MM: Evaluation of antibacterial and antidiarrhoeal activity of ethanolic extract of Feronia limonia The Pharma Innovation Journal 2015; 3(11): 50-54.
17. Pandavadra M and Chanda S: Development of quality control parameters for the standardization of acidissima L. leaf and stem. Asi Pac J Trop Med 2014; 7(1): 244-48.
18. Thirugnanasampandan R and David D: In-vitro antioxidant and cytotoxic activities of essential oil of Feronia elephantum Asian Pac J Trop Biomed 2014; 4(4): 290-93.
19. Pandit K, Mishra R, Brijesh S, Bhagwat A and Bhatt P: Lipid lowering activity of Feronia limonia leaf in Triton wr-1339 (Tyloxapol) induced hyperlipidemic rats. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6(8): 156-58.
20. Praveen D, Chowdary PR, Sajel S and Aanandhi MV: Muscle relaxant and antibacterial activity of leaf extracts of Feronia limonia. Journal of Chemical and Pharmaceutical Research 2015; 7(9): 809-12.
21. Wijewardana NA, Nawarathne SB and Wickramasingh I: Effect of various dehydration methods on proximate composition and retention of antioxidants in different fruit powders. International Food Research Journal 2016; 23(5): 2016-20.
22. Butsat S and Siriamornpun S: Effect of solvent types and extraction times on phenolic and flavonoid contents and antioxidant activity in leaf extracts of chinense C. International Food Research Journal 2016; 23(1): 180-87.
23. Umaiyambigai D, Saravanakumar K and Raj GA: Free radical scavenging activity, phenol and flavonoid contents of various solvent extracts from the Psydrax dicoccos (Gaertn). International Journal of Applied Research 2017; 3(1): 394-02.
24. Ahmad S, Arshad M A, Ijaz S, Khurshid U, Rashid F and Azam R: Review on methods used to determine Antioxidant activity. International Journal of Multi-disciplinary Research and Development 2014; 1(1): 35-40.
25. Raza A, Malook SA, Shahzad N, Qasrani SA, Sharif MN, Akram MN and Ali MU: Extraction of bioactive components from the fruit and seed of Jamun (Syzygium cumini) through conventional solvent extraction method. American-Eurasian J Agric & Environ Sci 2015; 15(6): 991-96.
    

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    How to cite this article:

    Srivastava R, Mishra N, Tripathi S and Mishra N: Effect of solvents on antioxidant activities of Feronia limonia fruit. Int J Pharm Sci & Res 2020; 11(7): 3385-91. doi: 10.13040/IJPSR.0975-8232.11(7).3385-91.

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