Optimization of Reference Standard Extract Preparation from Hedera helix L. by Response Surface Methodology and Single-Standard HPLC-DAD Quantification

[摘要] ObjectiveThis study optimized the extraction of a reference standard extract enriched in two bioactive triterpene saponins, hederacoside C and α-hederin, from Hedera helix L. leaves, and developed a high-performance liquid chromatography-diode array detection (HPLC-DAD) method based on a single standard to determine multi-components (SSDMC) for their simultaneous quantification.MethodsA D-optimal response surface methodology was applied to evaluate the effects of ethanol concentration, number of extraction cycles, and solvent-to-herb ratio on the extraction efficiencies of both compounds. The optimized extract was characterized by thin-layer chromatography and HPLC analysis. In addition, an HPLC-DAD method based on the SSDMC approach was developed, using hederacoside C as the single reference compound for the calculation of the relative conversion factor.ResultsThe optimal extraction conditions were determined to be 74% ethanol, three extraction cycles, and a solvent-to-herb ratio of 16:1, under which the predicted contents of hederacoside C and α-hederin were 3.95% and 1.79%, respectively. These predictions were in good agreement with the experimental values. The optimized procedure afforded a reference standard extract with a relatively simple chemical profile, in which hederacoside C and α-hederin were confirmed as the predominant constituents. Quantitative results obtained using the conventional external standard method and the SSDMC method showed good agreement.ConclusionThe optimized extraction procedure successfully produced a reference standard extract enriched in hederacoside C and α-hederin with good reproducibility. The developed SSDMC-based HPLC-DAD method demonstrated good applicability for the simultaneous quantification of both compounds and provides a practical approach for the quality control of H. helix preparations.

[发布日期] 2026-08-01

[关键词] conversion factor;D-optimal design;Hedera helix;SSDMC;reference standard extract