[Development of a quality control method for Polygalae Radix and exploration of its inter-column transfer strategy]

This study aimed to develop an ultra-performance liquid chromatography(UPLC) method for the quantitative analysis of xanthones and oligosaccharide esters in Polygalae Radix(PR), and to explore a retention-time-ratio-based strategy for inter-column method transfer. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C_(18) column(2.1 mm×50 mm, 1.7 μm) by gradient elution using 0.1% formic acid in water and acetonitrile with detection at 238 nm. The UPLC fingerprints of 31 batches of PR samples were established, with 21 common peaks marked and identified by UPLC-Q-TOF-MS~E. Through systematic methodological validation, a quantitative method was developed for the simultaneous determination of 7 bioactive components, namely sibiricose A_1, sibiricaxanthone B, polygalaxanthone Ⅷ, polygalaxanthone Ⅺ, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose, and tenuifoliside A. A quantitative analysis of multiple components by single marker(QAMS) method was established using polygalaxanthone Ⅲ as the internal reference substance, and its accuracy was compared with that of the external standard method(ESM). The feasibility of method transfer across different columns and instruments was also investigated. All 7 analytes showed good linearity(r>0.999) within their respective concentration ranges with mean recoveries of 94.31%-99.02%(RSD<2.0%, n=6). The relative error(RE) between QAMS and ESM was less than 4.0%, and the paired t-test indicated no significant difference between the two methods. Preliminary studies of method transfer on 9 C_(18) columns from 3 manufacturers on Waters UPLC systems of two models demonstrated that gradient time adjustment based on retention time ratio could maintain the elution behavior and separation characteristics of the target peaks, though the generalizability of this strategy requires further validation. The established UPLC-QAMS method is accurate and reliable for the quality control of PR. The inter-column method transfer strategy provides preliminary reference for the broader application of analytical methods.