Pharmacogn. Res., 2023; 15(2):267-276. https://www.phcogres.com Original Article Pharmacognosy Research, Vol 15, Issue 2, Apr-Jun, 2023 267 DOI:10.5530/pres.15.2.029 Copyright Information : Copyright Author (s) 2023 Distributed under Creative Commons CC-BY 4.0 Publishing Partner : EManuscript Tech. [www.emanuscript.in] Analytical Quality by Design (AQBD) Assisted Development and Validation of HPTLC Method for Estimation of Rottlerin in Topical Patch Formulation Kaumudee Bodas 1 , Vaibhav M Shinde 1, *, Deshmukh Vishal 2 , Deshmukh Sheetal 2 1 Centre for Advanced Research in Pharmaceutical Sciences, Poona College of Pharmacy, Pune, Maharashtra, INDIA. 2 Bharati Vidyapeeth (Deemed to be University), Yashwantrao Mohite Institute of Management, Karad, Maharashtra, INDIA. ABSTRACT Background: Rottlerin, a natural polyphenolic ketone is a major constituent of Mallotus philippensis (Lam.) Mull. Arg. plant. Although, less explored it has diverse array of therapeutic applications. Objectives: The current work aimed to develop novel, simple, speedy, accurate, potent High Performance Thin Layer Chromatography Method (HPTLC) to separate, and quantify rottlerin from the herbal topical patch In-House formulation. Materials and Methods: This was the foremost attempt to develop and validate HPTLC method using Analytical Quality by design (AQbD) approach. Box Behnken design (BBD) coupled with response surface methodology approach was followed for the systematic optimization. The end results of toluene content, chamber conditioning time, and solvent front on R f value of rottlerin was studied. The optimised method was validated as per ICH guidelines. Results: For HPTLC, stationary phase used was aluminium plates 60F 254 coated with silica gel as. AQbD optimised conditions were mobile phase proportion namely toluene: ethyl acetate: methanol: ammonia (5:4:2:0.2), development distance 80 mm, band width 6mm, and chamber saturation time 10 min. The method generated well resolved and compact bands at R f of 0.41±0.05. The developed method exhibited high accuracy, precision, robustness, and recovery. The quantity of Rottlerin in developed herbal patch was found to be 134.8 ng/band. Conclusion: In a nutshell, the present study undeniably vouches for a QbD based method development approach for Rottlerin and can be extrapolated to estimate it in other herbal extracts, or marketed formulations. Keywords: Rottlerin, Analytical Quality by design (AQbD), HPTLC, Mallotus philippensis (Lam.) Mull. Arg., Topical patch. INTRODUCTION Mallotus philippensis (Lam.) Mull. Arg. commonly known as Kamala tree, is one of the well–known but less explored plants in traditional systems of medicines. It is very well described in ancient Indian literature Charaka Samhita and Sushruta Samhita. Te fruits of the plant are most common element of various Ayurvedic and Unani formulations. [1] It has wide array of therapeutic applicability such as Dermatological disorders, [2] Digestive disorders, [3,4] Helminthic disorders, [5,6] Infammatory disorders, [7,8] Microbial infections, [9,10] Psychological disorders, [11] Reproductive disorders, [12] Respiratory disorders, [13] in vitro cytotoxicity, [14] etc. Rottlerin, a natural polyphenolic ketone is the main bioactive component present in pericarp of Kamala plant. It is a 5,7 dihydroxy–2,2–dimethyl-6-(2,4,6 trihydroxy-3methyl-5acetyl benzyl)-8- cinnmoyl–1,2- chromene as displayed in Figure 1, also known as mallotoxin. Recent scientifc data demonstrated the potential of rottlerin to modulate variety of molecular targets such as protein kinase C (PKC),5’- AMP activated protein kinase, p38 regulated kinase, C AMP dependent kinase, Casein kinase II, Glycogen synthase kinase 3 beta, AKT/PKB, and calmodulin– dependent kinase etc. [15] Also it can modulate Human Interleukin–6 (IL 6), Interleukin–1 beta (IL-1β), NF–Kappa B (NF-κB), transforming growth factor–beta type I receptor (TGF1 -β),Insulin–like Growth Factor Receptor (IGF 1R), Fibroblast Growth Factor–1 (FGF–1), Matrix Metalloproteinase (MMP 9). [16] Due to diverse therapeutic pertinence, rottlerin represents promising drug candidature to treat various diseases. But it lacks scientifc evidence due to unavailability of precise analytical method to ensure safety, efcacy and quality of rottlerin, and its formulations. As per literature analytical approaches for Received: 05-11-2022; Revised: 16-12-2022 ; Accepted: 11-01-2023. Correspondence: Dr. Vaibhav M. Shinde Centre for Advanced Research in Pharmaceutical Sciences, Poona College of Pharmacy, Pune, Maharashtra–411038, INDIA. Email : vaibhavshinde847@gmail.com