上海中医药杂志2026，Vol.60Issue(1)：64-72,81,10.DOI:10.16305/j.1007-1334.2025.z20250905005
黄芪中皂苷类成分UHPLC‑ELSD分析方法的建立与应用
Development and application of an UHPLC‑ELSD method for analysis of saponins in Astragali Radix
摘要
Abstract
Objective To establish a simple,rapid and practical quantitative analysis method for multiple astragalosides,including astragaloside Ⅰ(ASⅠ),isoastragaloside Ⅰ(Iso-ASⅠ),astragaloside Ⅱ(ASⅡ),isoastragaloside Ⅱ(Iso-ASⅡ),astragaloside Ⅲ(ASⅢ),astragaloside Ⅳ(ASⅣ)and cycloastragenol(CA)in Astragali Radix and to lay a foundation for improving the quality standard of Astragali Radix.Method The sample pretreatment method was optimized through single-factor experiments and response surface methodology(Box-Behnken design),focusing on three key parameters:extraction time,extraction solvent,and solid-to-liquid ratio.Quantitative analysis of astragalosides in Astragali Radix was performed using ultra-performance liquid chromatography coupled with an evaporative light scattering detector(UHPLC-ELSD).The chromatographic separation was achieved on a Waters HSS T3 column(2.1 mm×100 mm,1.7 μm)using a mobile phase consisting of A(0.1%formic acid aqueous solution)and B(acetonitrile)and eluted at a flow rate of 0.3 mL/min by the following gradient program:0-1 min,28%B to 35%B;1-6 min,35%B to 35%B;6-13 min,35%B to 48%B;13-17 min,48%B to 48%B.The injection volume was 8 μL,and the column temperature was maintained at 30℃.The evaporator temperature and nebulizer temperature were set at 60℃and 70℃respectively with a gas flow rate of 1.6 SLM.Results The optimal sample pretreatment conditions were determined as follows:2 h of heated reflux extraction using 70%methanol as the solvent and a solid-to-liquid ratio of 1∶60.The established quantitative method demonstrated excellent specificity.The precision validation showed RSD values ranging from 0.30%to 2.89%,and the recovery rates ranged from 95.65%to 105.18%.The RSDs for repeatability and 48-hour sample stability were within 2.92%and 2.83%,respectively(no repeatability or stability data were available for CA in Astragali Radix samples due to its content being below the detection limit).All validation results complied with acceptance criteria for quantitative analysis.Quantitative results revealed significant positive correlations among astragalosides,and their contents were related to geographical origin and growth years.The content of astragalosides followed the order:ASⅠ＞ASⅡ＞Iso-ASⅠ≥ASⅣ＞Iso-ASⅡ＞ASⅢ,with the level of CA below the detection limit.Conclusions A simple,rapid,efficient,and well-separated quantitative method is established,enabling accurate and simultaneous determination of multiple astragalosides.This approach provides a foundation for selecting quality markers among astragalosides and establishing scientific quality standards for Astragali Radix.关键词
黄芪/黄芪皂苷/超高效液相色谱仪联用蒸发光散射检测器/质量控制/中药研究Key words
Astragali Radix/astragalosides/UHPLC-ELSD/quality control/research of Chinese materia medica引用本文复制引用
JIANG Hui,WANG Changhong,LIU Wei,CHENG Xuemei,ZHENG Liming,MAO Xinmin,ZHAO Lu,MA Xuan,JI Zhihong,LI Jianguang..黄芪中皂苷类成分UHPLC‑ELSD分析方法的建立与应用[J].上海中医药杂志,2026,60(1):64-72,81,10.基金项目
国家重点研发计划项目(2022YFC3501701) （2022YFC3501701）
克拉玛依市重点研发计划项目(2025BA0090) （2025BA0090）
新疆维吾尔自治区科技计划项目(2024LQ03007) （2024LQ03007）
