Abstract
Pharmacological activity of Aloe vera is known for hundreds of years, but a precise and well-established characterization method for all kinds of pharmaceutical formulations is still a challenging task. In the present study, a simple, user- friendly, sensitive, precise, accurate, robust and reproducible method has been developed based on reverse phase-high performance liquid chromatography (RP-HPLC). The RP-HPLC method has been developed, standardized and validated utilizing the Aloe marker compounds viz., Aloin A, Acemannan and Aloe-emodin that is present in various Aloe vera varieties. The total polyphenolic content (TPC) and total flavonoid content (TFC) were estimated spectrophotometrically and an in-vitro antioxidant study was also performed to standardize the potential of Aloe vera using assays viz. DPPH (2,2- diphenyl-1-picrylhydrazyl radical scavenging), NO (nitric oxide) scavenging potential, FRAP (ferric reducing antioxidant power) and TAC (total antioxidant capacity). Simultaneously, tocopherol acetate was also estimated in commercially manufactured pharmaceutical products with the help of the previously standardized HPLC method in our laboratory. Separation of the singular active ingredient of Aloe vera was achieved by using an isocratic mode of acetonitrile and water (70:30 v/v) by using a reversed-phase C18 column as stationary phase in a high-performance liquid-chromatography system employing photodiode array detector (PDA plus detector) with a flow rate of 1.0 ml/min. The detection limit of active compound of Aloe species was found to be in the range of 0.00020 to 0.00051 µgL-1 (20 µL injection of each for five times). The quantitative method of Aloe vera extracts standardized vis-à-vis both with peel (AL-P) and without peel (AL-WP) form gives robust, precise (% RSD 1.13-3.84) and accurate results. This method is suitable for the detection of major pharmacologically active compounds present in Aloe vera-based pharmaceuticals and nutraceuticals.
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