Evaluation of Antioxidant and Total Phenolic Content of
Family Rosaceae as Major Indigenous Fruit Type of
Kargil Ladakh in Trans Hiamalaya
Mohd Sadiq Lone1 and Mohd Ishaq2
1Bhagwant University,
Sikar Road Ajmer, Rajasthan, INDIA. 2Department of Chemistry,
Govt. Degree College, Kargil Ladakh, J & K, INDIA. email:[email protected].
(Received on: October 23, 2016)
ABSTRACT
Plants grown in Kargil Ladakh are known to respond extremely cold, dry weather and harsh environmental conditions which are extreme by any other standard by synthesing phytochemicals to fight adverse for their survival, this out lined the necessicity of determining their content in fruits of kargil in trans himalayas.
Three species of family rosaceae viz Prunus armeniaca, Prunus siberiaca & Malus domestica grown as major indegineous fruit under similar cultural practices in Kargil Ladakh is taken into consideration for antioxidant & total phenolic content evaluation.
The total phenolic content was determined by using Folin-ciocaltue assay, Ferric reducing antioxidant potential (FRAP) to determine antioxidant content was measured spectrophotometrically.
The results of the fruits showed that the methanolic extract of P.armeniaca exbhit heighest TPC(68.2mgGAE/100g) fowellod by P.siberica (67.1mgGAE/100g) & M.domistica (65.7mgGAE/100g). Water extract of P.armeniaca also exbhit slightest highest TPC (44.2mgGAE/100g) as compare to other two P.siberace (42.1mgGAE/100g) & M.domistica (40.1mgGAE/100g). The mean heigest value of FRAP was observed in methanolic extract of of P.armeniaca (123.6µg/ml) followed by M.domistica (122.5µg/ml) and P.siberiaca (122.1µg/ml) water extract of P.armeniaca showed heighest FRAP value ( 69.1 µg/ml) followed by M.domestica (68.3µg/ml) P.siberiaca (68.2 µg/ml).
INTRODUCTION
Antioxidants are substances that are capable of suppress the oxidative damage of proteins, lipids and nucleic acids oxidative damage on cellular level is caused by highly reactive oxygen species which include reactive free radicals such as peroxides, superoxide’s etc. antioxidant deactivate free radicals by breaking chain propagation suppressing formation of free radicals with high affinities by reducing hydrogen peroxide binding to metal ions and quenching superoxide’s and singlet oxygen1. Natural products for food and nutritional supplements have gained increased attention in recent years in this context there is an increasing interest in the beneficial health effects of plants derived antioxidant rich diets and lower incidences of degenerative diseases including cancer, heart diseases, inflammation, immune system decline, brain dysfunction2-3. In addition to their important role in biological system antioxidants play a vital role in food industry and extend the life span of foods containing oxidizable lipids such as spices, nut, flavourings, vegetable oils4.
Fruits of family rosaceae (P.armeniaca, P.siberiaca, M.domistica) apricots and apples are widely consumed in many parts of the world the total world production of apricot is about 2.6 million tons with Turkey as leading country5. The fruit contains different types of phytochemicals such as polyphenols, vitamins, carotenoids etc. which contribute to their nutritional value and tast6 the apricot seed represents about 15% of fruit and kernel represent about 34% of seed7. In family rosacae another important species of fruit is Malus domistica (apple) widely consumed through out the world, about 80 million tons of apples were grown worldwide in 2013 and China produce almost half of this total, USA being second. Apple trees are large if grown from seed but small if grafted on to roots and are generally propagated by grafting, although wild apples grow readily from seed, trees of apple are prone to a number of fungal, bacterial and pest problems which can be controlled by a no of organic and non organic means since fresh apricot and apple fruits are good source of antioxidant8 such as chlorogenic acid, neo chlorogenic acid, ferulic acid, beta carotene and gama carotene6. The present research work describe evaluation of TPC and antioxident potental by using two different assay using folin reagent and FRAP in three different species of family roseasace indigenous to Kargil in trans himalayas and to explore its importance to further improve the quality and standard of fruits of this region which constitute an important economic source for this area.
MATERIAL AND METHODS/ EXPERIMANTAL
A) Sample collection and geography of location
practices were performed. The selection of the cultivars was based on their importance in the region Halman Khantachuli and Saspol are the most important cultivars predominantly grown on limited scale in the region, the fruit quantative tracts such as fruit wt. fruit dia. were determined at the time of harvest and other parameters such as TSS,TA, PH and TDS of fruit were determined after harvest.
B) Sample preparation
The fruit cultivars in this study were apple (M.domistica) and apricot (P.armeneca, & P. siberiaca) of family rosaceae, the fruits were harvested at the technological ripe stage in the commercial plantation in Hardass village of Kargil a major apricot producing site, two cycles of extraction were performed ie with methanol and water, sample preparation and juice extraction of fruits into pieces and mixed to make a homogeneous pool, fruit juice of (2.5ml) were extracted for 60 minutes with 100ml of methanol in a capped bottle in an orbital shaker at room temp again fruit juice of 2.5 ml was extracted for same time with water.
C) Total Phenolic Content
TPC was measured using the Folin - ciocaltue colorimetric method as described (Gao ohlande, jeppsson 2000) the methanolic fruit extract ( 10 µL of 0.1 mg/ml) was mixed with 20µL of folin -ciocalteu reagent and 200 ml of water and incubated at room temp for 30 minute, following the addition of 100µl of 20% sod. carbonate to the mixture, TPC was determined after one hour of incubation at room temp. the absorbance of the resulting color was measured at 765nm using spectrophotometer, TPC was expressed as mg GAE /100g fruit wt.
D) Antioxidant capacity
Ferric Reducing antioxidant potential (FARP) assay FRAP reagent 300µ mol sod acetated buffer (3.1g CH3 COONa and 16ml CH3 COOH) at PH 3.6 10 ml 2,4,6,_ tri ( 2- pyridyl) –s- trizine (TPTZ) Sol. in 40 u ml HCL and 20 µ ml fecl3 6H2O sol. The fresh working sol. Was prepared by mixing 25 ml aceted buffer 2.5 ml TPTZ and 2.5 ml feCl3. 6 H2O sol. fruits extracted ( 15 µL of 0.1 mg /ml ) was allowed to react with 285 µL of FRAP sol. For 30 minutes in dark observation of colored was measured at 593nm. the results expressed as µg / ml Feso4.
RESULT AND DISCUSSION
Total phenolic content
of organic acid as observed from PH and acidity, organic acid reacts with folin ciocaltue reagent in the TPC determination.
Anti oxidant content
Antioxidant activity by FRAP method in the present study was done with water extraction followed by methanol, it was observed that the second extraction with methanol retained significally higher TAC in the sample. The mean highest value of FRAP was observed in methanolic extract of p. armeniaca (123.6 µg /ml) followed by M. domestica (122.5µg /ml) and p. siberica (122.1µg /ml) . in water extract also p. armenaica showed highest TAC with FRAP value ranging from (69.1µg /ml) followed by m.domistica (68.3µ/ml).
Table S1- Shows antioxidant and total phenolic content of Rosacea collected from major fruit producing location Hardass Kargil in trans – Himalayas.
S. No property unit p.armenica p.siberica M.Domestica
1 TPC(w) mg GAE/100g 44.2 42.1 40.1
2 TPC(m) mg GAE/100g 68.2 67.1 65.7
3 FRAP(w) µg/ml FeSO4 69.1 68.2 68.3
4 FRAP(m) µg/ml FeSO4 123.6 122.1 122.5
Table S2- Shows total antioxidant content by FRAP in three species of Rosaceae (methonal & water extract) collected from a major fruit producing site Hardass Kargil in trans Himalayas.
s.no Property Unit P. armenica P.siberica M.domestica
FRAP(w) µg/ml 69.1 68.2 68.3
FRAP(m) µg/ml 123.6 122.1 122.5
Table S3- Shows the total phenolic content in three species of Rosaceae (methanolic and water extract) collected from a major fruit producing site Hardass kargil in trans Himalayas.
S.No. Property Unit P.armeniaca P.siberiaca M.domistica
1 TPC(w) mgGAE/100g 44.2 42.1 40.1
2 TPC(m) mgGAE/100g 68.2 67.1 65.7
CONCLUSION
The above analysis revels that p. armeniaca contained extensive nutritional component as well as highest total phenolic content among the three species of rosaceae. It was also observed that methanol is better solvent than water because it dissolved more antioxidant as observed above, and FRAP assay is best among the two assays ln determination of antioxidants. As species of fruit under examination are widely grown in harsh environmental conditions of Kargil Ladakh in trana Himalaya which is extrem by any other standard, our focus in this study remains in the evaluation of antioxidant & total phonlics of indigenous fruits of this regions and observed valus are satisfactory and further improved by applying more and more new techniques and methods of cultivation, also Govt should take constructive measures in further improving these values by involving different agencies In fruit cultivation in these regions.
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