JOURNAL OF NATURAL REMEDIES DOI: 10.18311/jnr/2022/31350 Article Received on: 08.07.2022 Accepted on: 11.11.2022 Revised on: 19.10.2022 1. Introduction Ayurvedic formulations provide basic healthcare treatment for patients with several ailments 1 . As the use of ayurvedic formulations increases, so do increases concerns about their safety and efcacy. Hence, it is essential to provide more information to properly understand the risks related to the use of these products 2 . Te World Health Organization also emphasizes quality control of ayurvedic formulations by using modern techniques and providing suitable parameters and standards. Te quality of herbal plants and their medicines is mainly defned by the content of efective biomarkers present in them 3,4 . For the proper defnition of this term and properly evaluating the dynamic nature of various biomarkers, many chromatographic and spectroscopic techniques were developed. Quality control of ayurvedic formulations is an efcient procedure for pharmaceuticals to ensure the purity and uniformity of marketed pharmaceutical products. So, diferent tools and techniques are implied to ensure quality 5 . Te HPTLC method is useful in a variety of pharmaceutical analysis, including qualitative and quantitative compound and impurity determination, assays, semi-quantitative limit tests, and degradation studies 6 . High-Performance Tin Layer Chromatography (HPTLC) is a chromatographic technique that helps in the proper separation, detection and quantifcation of active constituents present in plant extracts and their related formulations 7 . HPTLC has a better ability to separate multi-components and is really easy with various coloured chemical constituents. Many sensitive colour reagents can also be used in HPTLC for the detection of spots 8 . Amrtadi churna is a polyherbal formulation that aids in the treatment of hyperacidity (a type of pitta dosha) and many other geriatric diseases. Amrtadi churna consists of one part each of ‘Guduchi’, ‘Amla’ and ‘Gokshur’ 9 . RESEARCH ARTICLE HPTLC Method Development and Validation for Simultaneous Estimation of Berberine, Ellagic Acid and Ferulic Acid in Amrtadi Churna Nikita Thorat, Tanvi Dodiya* and Disha Prajapati Parul Institute of Pharmacy and Research, Parul University, Limda, Waghodia, Vadodara - 391760, Gujarat, India; pateltanvi1982@gmail.com Abstract The objective of the current study was to develop a new simple and precise High Performance Thin Layer Chromatography (HPTLC) method for standardization of three biomarkers i.e., berberine, ellagic acid and ferulic acid in Amrtadi churna. Amrtadi churna is mainly used for hyperacidity and contains dried plant parts of Gokshur, Amla and Guduchi. The method was developed and validated using precoated silica gel at 60 F 254 as the stationary phase and toluene:ethyl acetate:formic acid:methanol (6:6:1.6:0.4, v/v/v/v) as the mobile phase. The detection and quantification were performed at 319 nm and the R f value obtained was 0.35±2 for berberine, 0.5±2 for ellagic acid and 0.74±2 for ferulic acid. The method was validated as per ICH guidelines in terms of linearity, precision, specificity, accuracy and robustness. Keywords: Amla, Gokshur, Guduchi, ICH, Standardization