A multi-technique analytical strategy for the standardization of <i>Boswellia serrata</i> : RP-HPLC quantification employing relative response factors, HPTLC fingerprinting, LC-MS/MS characterization

Boswellia serrata is a widely recognized medicinal plant whose therapeutic properties are primarily attributed to boswellic acids, making their reliable characterization and quantification essential for its quality assessment. This study established an integrated analytical strategy for the standardization of Boswellia serrata through marker isolation, chromatographic fingerprinting, quantitative analysis, and compound characterization. A key constituent was isolated by column chromatography and identified as elemadienoic acid by LC-MS/MS analysis. Additional characterization enabled identification of major boswellic acids, along with two markers tentatively assigned as 3α- and 3β-hydroxytirucalla-7,24-dien-21-oic acid. HPTLC fingerprinting provided a rapid tool for comparative evaluation of different Boswellia matrices. A validated RP-HPLC method enabled simultaneous quantification of six major boswellic acids. The method demonstrated excellent linearity (r2 ≥ 0.9999), high precision (%RSD ≤ 1.43%), acceptable accuracy, specificity, robustness, and solution stability. A relative response factor (RRF) strategy using 11-keto-β-boswellic acid (KBA) as a single reference standard produced results comparable to conventional multi-standard calibration while reducing dependence on multiple reference standards, offering a cost-effective approach for routine quality control. The integration of marker isolation, HPTLC fingerprinting, RRF-based RP-HPLC quantification, and LC-MS/MS characterization provides a comprehensive and reliable framework for supporting improved quality assessment of different boswellia matrices.