UHPLC with dual detection fingerprinting and determination of multi-indicator components of Fructus Liquidambaris

WANG Chang-yu; ZHANG Xin-hua; CHAI Rui-ping; ZHANG Lu; TAO Xin-yu; ZHANG Hong-yang; HU Ping

doi:10.20251/j.cnki.1003-3734.2025.02.012

Abstract：Objective:To establish an UHPLC dual detection fingerprint for Fructus Liquidambaris and develop a method for determining the content of three phenolic acid compounds and three triterpenoid compounds in Fructus Liquidambaris to comprehensively reflect the overall distribution of its chemical components.Methods:The fingerprint analysis was conducted using UHPLC coupled with a UV detector and a charged aerosol detector(UHPLC-UV-CAD).Thermo Acclaim RSLC C18(100 mm×2.1 mm,2.2 μm)column was used,with acetonitrile-0.1%formic acid aqueous solution as the mobile phase in gradient elution procedure 1,and the detection wavelength was 254 nm.The content determination of protocatechuic acid,gallic acid,and paeonol was performed using UHPLC-PDA at wavelengths of 254 nm,259 nm,and 274 nm in gradient elution procedure 2.The content determination of betulonic acid,oleanolic acid,and ursolic acid was carried out using HPLC-CAD,utilizing a Sunrise C30 column(250 mm×4.6 mm,5.0 μm)with gradient elution procedure 3.Results:A total of 16 common peaks were identified in the fingerprint,demonstrating good precision,repeatability,and stability of the method.The similarity of the fingerprints of 18 batches of Fructus Liquidambaris was all≥0.935,indicating a high similarity in chemical composition.The standard curves of the six indicator components showed good linearity(R2≥0.9996),with average spiked recoveries ranging from 93.7%to 103.8%,and the RSD of recovery rates were≤3.07%.The mean contents of protocatechuic acid,gallic acid,paeonol,betulonic acid,oleanolic acid,and ursolic acid in 18 batches of Fructus Liquidambaris were 0.0041%,0.0064%,0.0154%,0.299%,0.310%,and 0.219%,respectively.Conclusion:The method established in this study is accurate,sensitive,and reproducible,capable of reflecting the chemical components in Fructus Liquidambaris more comprehensively.Among these,the triterpenoid components significantly contribute to the intrinsic quality of Fructus Liquidambaris.