Box–Behnken Design Based Optimisation of HPTLC Method for Analysis of Jatrorrhizine from Tinospora cordifolia Roots with Antidiabetic Activity Assessment

Pharmacognosy Research,2025,17,4,1382-1394.DOI:10.5530/pres.20252320Published:August 2025Type:Original ArticleAuthors:Neeraj Kumar, Akanksha A, Rama Tyagi, Shubham Singh, Mohd Aqil, Abul kalam Najmi, Mohd Vaseem Ismail, and Mohd. Mujeeb Author(s) affiliations:Neeraj Kumar1, Akanksha1, Rama Tyagi2, Shubham Singh1, Mohd Aqil3, Abul kalam Najmi4, Mohd Vaseem Ismail5, Mohd. Mujeeb1,* 1Department of Pharmacognosy and Phytochemistry, School of Pharmaceutical Education and Research (SPER), Jamia Hamdard, New Delhi, INDIA. 2Department of Pharmacognosy and Phytochemistry, Galgotia University, Greater Noida, INDIA. 3Department of Pharmaceutics, School of Pharmaceutical Education and Research (SPER), Jamia Hamdard, New Delhi, INDIA. 4Department of Pharmacology, School of Pharmaceutical Education and Research (SPER), Jamia Hamdard, New Delhi, INDIA. 5School of Pharmaceutical Education & Research (SPER), Jamia Hamdard, New Delhi, INDIA. Abstract:Introduction: Tinospora cordifolia root (T. cordifolia) is a well-known Ayurvedic herb with immune-boosting, anti-inflammatory, and antioxidant properties. It contains Jatrorrhizine (JZ), a bioactive alkaloid with antimicrobial, hepatoprotective, and metabolic health benefits, similar to berberine. Objectives: The study aimed to evaluate the JZ content in T. cordifolia root obtained via different extraction method and to optimise the High-Performance Thin Layer Chromatography (HPTLC) method via Response Surface Methodology (RSM). The extracts were assessed for in vitro test evaluation. Materials and Methods: A Box-Behnken Design (BBD) of RSM, a mathematical and statistical method, was applied to optimize the impact of independent variable on the response. Three independent variables like the different concentration of the mobile phase were studied. The in vitro analysis was also done. Results: The content of JZ was quantified using the HPTLC densiometric method. HPTLC confirmed the presence of JZ with a prominent band at an Rf value of 0.70±0.008 using a mobile phase of chloroform: toluene: methanol: formic acid (7:4:2:0.2 v/v). The in vitro analysis of the extract showed a significant result and was found to be α-amylase in TRM 27.71±0.70 and α-glucosidase in TRM 43.45±1.31. Conclusion: This work examines various extraction procedures, including Soxhlet, maceration and reflux, and introduces a novel methodology to enhance extraction efficiency, advance the phytochemical analysis of JZ. This suggests that T. cordifolia may have therapeutic potential in diabetes management. Keywords:HPTLC, Jatrorrhizine, Response surface methodology, α-Amylase, α-GlucosidaseView:PDF (1.03 MB)