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DEVELOPMENT AND VALIDATION OF AN HPTLC METHOD FOR BAKUCHIOL QUANTIFICATION IN PSORALEA CORYLIFOLIA SEEDS USING CENTRAL COMPOSITE DESIGN

MOHD AMIR 1, DEEPAK KHURANNA 2, WASIM AHMAD 3, SHOWKAT R MIR2, MOHAMMAD RAISH 4, SHEIKH MANSOOR5, AJAZ AHMAD 6, MOHD MUJEEB2*

1Department of Natural Products & Alternative Medicine, College of Clinical Pharmacy, Imam Abdulrahman Bin Faisal University, Dammam, 31441, Saudi Arabia
2 School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi-62, India
3 Department of Pharmacy, Mohammed Al-Mana College for Medical Sciences, Safaa, Dammam 34222, Saudi Arabia
4 Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia
5 Department of Plant Resources and Environment, Jeju National University, 63234, Republic of Korea
6 Department of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh 11451, Saudi Arabia

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Bakuchiol (meroterpenic phenol) is a well-known active constituent present in Psoralea corylifolia Linn (Leguminosae family). The aim for the current study was to determine, develop and validate a new, simple, selective, precise, and accurate highperformance thin layer chromatographic (HPTLC) method for the assessment of bakuchiol in different extracts of P. corylifolia. A pre-coated aluminium plate with silica gel 60 F254 was used for the thin layer chromatographic separation of bakuchiol, eluted with toluene: ethyl acetate: acetic acid (10:1.0:0.1; v/v/v). The plate was subsequently dried, and a Camag TLC scanner III was used to perform densitometric scanning at 262 nm. Selected mobile system gave well defined and sharp chromatogram of bakuchiol at Rf value of 0.48 ± 0.07. Linearity, accuracy, precision, specificity, limit of detection (LOD), and limit of quantification (LOQ) were among the ICH guidelines parameters that were used in the method development and validation. Additionally, a central composite design (CCD) was used in robustness testing. With a fine correlation value (R 2 ) of 0.999, an excellent linear association between concentrations (200 - 2000 ng/band) and areas of the chromatogram was recorded. Intraday and inter-day precision evaluations showed the relative standard deviation 0.68 - 2.77. The percentage of recovery was found to be 98.42% - 101.91%. Results from recovery studies demonstrated outstanding reproducibility and dependability of the method. The LOD and LOQ were determined to be 181.74 ng/spot and 550.74 ng/spot, respectively. The selected factors evaluated in the robustness testing by CCD were found to have a significant effect on the peak area. Among all extracts, the methanolic extract showed maximum concentration of bakuchiol. HPTLC-based identification and quantification of bakuchiol in plant species as well as other plant based and modern medicine formulations may now be done with fresh insights provided by validated standards-based methodologies.