eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2026-05-11 23 2 58891 article Daksha Lalit Attarde 1 Sarika Tanaji Nikam 2 Komal Bhausaheb Dawange 1 Department of Pharmacognosy, Mahatma Gandhi Vidyamandir’s Pharmacy College, Nashik, India. Department of Pharmacognosy, Dr. D.Y. Patil Institute of PharmaceuticalScience & Research, Pune, India. The growing prevalence of oxidative stress-associated disorders highlights the need for standardized herbal nutraceuticals with proven antioxidant potential. The present study aimed to develop and HPTLC standardize an Ayurvedic Avleha-based nutraceutical formulation containing extract of Manilkara zapota fruit. The extract exhibited significant phytochemical richness, with total polyphenolic content of 290 mg/g (gallic acid equivalent), total flavonoid content of 31.06 mg/g (rutin equivalent), and total flavonol content of 2.5 mg/g (rutin equivalent). Antioxidant activity assessed by DPPH assay showed an IC₅₀ value of 5180 µg/mL for the extract, compared to 4.12 µg/mL for ascorbic acid. Among the developed batches, Batch III showed optimal physicochemical characteristics and was selected for further evaluation. HPTLC analysis using a validated method with mobile phase Toluene: Ethyl acetate: Formic acid: Methanol (6:6:1.6:0.4 v/v/v/v) at 275 nm revealed well-resolved peaks with Rf values of 0.39 and 0.29 for gallic acid and ellagic acid, respectively. Simultaneous estimation showed that gallic acid and ellagic acid contents were 0.6312% w/w and 0.6918% w/w in the extract, and 0.09421% w/w and 0.04420% w/w in the formulation, respectively. The method was validated as per ICH guidelines and found to be linear, precise, accurate, and robust, supporting its suitability for routine analysis. https://www.biotech-asia.org/vol23no2/development-and-hptlc-standardization-of-an-ayurvedic-avleha-based-nutraceutical-formulation-of-manilkara-zapota-fruit-extract-with-simultaneous-estimation-of-gallic-acid-and-ellagic-acid/ Avleha formulation; Ellagic acid; Gallic acid; HPTLC; Manilkara zapota; Sapodilla