Abstract
Objective: The objective of the present study was to develop an HPLC analytical method and to perform in vivo study of Emblica officinalis and Aegle marmelos extracts for antioxidant and antidiabetic activity.
 Methods: The phytochemical analyses, total phenolic content (TPC), TLC, DPPH assay were performed for freeze-dried Emblica officinalis fruits aqueous extract (EOFAE) and Aegle marmelos leaves ethyl acetate extract (AMLEAE). The active constituents present in both extracts were estimated by using HPLC system having Hibar® C18 column [250 x4.6 mm, 5 µm] and UV detector (264 nm). A gradient mobile phase (acetonitrile and water with 0.1% trifluoroacetic acid) was used at a flow rate of 0.8 ml/min. In vivo antioxidant, antidiabetic activity of both extracts was conducted on male albino Wistar rats for 21 d in streptozotocin-induced diabetic rats (42 rats; n=6). The antidiabetic activity was measured by blood glucose level and biochemical parameters i.e. total cholesterol, triglycerides and total protein. Oxidative stress was measured by antioxidant biomarkers i.e. SOD, GSH, lipid peroxidation by thiobarbituric acid reactive substances method on the liver of the experimental rat.
 Results: Tannins, saponins, carbohydrate, glycosides are found in EOFAE; coumarins and flavonoids are found in AMLEAE and quinones, phenols are present in both extracts. The values of TPC present in standard gallic acid, EOFAE and AMLEAE were found to be 485.7, 315.6, 300.7 mgGAE/g, respectively. Rf values obtained by TLC of EOFAE and AMLEAE were found to be 0.41 and 0.50, respectively. The values of % inhibition shown by EOFAE and AMLEAE in DPPH assay were found to be 97.8%±2 and 95.2%±2, respectively. The values of retention time of EOFAE and AMLEAE by HPLC analysis were found to be 4.59 and 5.28 min, respectively. Histopathological examination of the liver was revealed that low dose EOAM (containing of EOFAE 250+AMLEAE 250 mg/kg body weight) administered once a daily for 21 d showed significant activity (P˂0.001) with biochemical parameters and antioxidant biomarkers.
 Conclusion: The present study showed that the EOFAE and AMLEAE treated group III with (EOAM) low dose of 500 mg/kg body weight has potent antioxidant and antidiabetic activity.
Highlights
World Health Organisation (WHO) states that an estimated 1.6 million deaths were directly caused by diabetes in 2015 and projects that this disorder will be 7th leading cause of death in 2030 [1]
Proposed project involves the co-administration of aqueous extract of Emblica officinalis fruits and ethyl acetate extract of Aegle marmelos leaves for antioxidant and antidiabetic activity
Total phenolic content The total phenolic contents of gallic acid, Emblica officinalis fruits aqueous extract (EOFAE) and Aegle marmelos leaves ethyl acetate extract (AMLEAE) were found to be 485.7, 315.6 and 300.7 milligrams of gallic acid equivalent per gram of dried extract, respectively. These results indicated that EOFAE and AMLEAE have sufficient phenolic content for antioxidant activity
Summary
World Health Organisation (WHO) states that an estimated 1.6 million deaths were directly caused by diabetes in 2015 and projects that this disorder will be 7th leading cause of death in 2030 [1]. Diabetes mellitus is a chronic metabolic disorder caused by inherited and/or acquired deficiency in production insulin by the pancreas or by the ineffectiveness of the insulin produced. Such a deficiency results in increased concentrations of glucose in the blood, which in turn damage many of the body’s systems, in particular, the blood vessels and nerves [5]. Is known as “Amla” traditionally, belonging to the family Euphorbiaceae [12] It is one of the most important plants of Ayurveda and traditional Indian medicine [11]. Proposed project involves the co-administration of aqueous extract of Emblica officinalis fruits and ethyl acetate extract of Aegle marmelos leaves for antioxidant and antidiabetic activity
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