快速溶剂萃取-超高效液相色谱法测定稳心颗粒中皂苷类成分

目的: 建立快速溶剂萃取-超高效液相色谱法,用以测定稳心颗粒中三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁的含量. 方法: 利用乙醚脱脂,水饱和正丁醇提取制剂中皂苷;采用Agilent Eclipse Pluse C₁₈色谱柱(2.1 mm×50 mm,1.8 μm)分离,以乙腈(A)-水(B)为流动相,梯度洗脱(0~10 min,81%B;10~17 min,81%B→45%B;17~18 min,45%B→81%B;18~25 min,81%B),流速0.3 mL·min^-1,柱温25℃,检测波长203 nm.与普通提取方法比较,提取时间由14 h缩短到25 min,大幅度提高样品处理效率. 结果: 三七皂苷R₁、人参皂苷Rg₁和人参皂苷Rb₁进样量分别在0.084~0.84 μg(r=0.999 9)、0.201 6~2.016 μg(r=0.999 8)、0.208 4~2.084 μg(r=0.999 9)范围内呈良好线性关系,平均加样回收率(n=6)分别为98.4%(RSD=1.1%)、98.2%(RSD=0.47%)、97.9%(RSD=1.3%).样品中总皂苷含量为2.65 mg·g^-1,与中国药典方法测定结果(2.63 mg·g^-1)基本一致.采用快速溶剂萃取-超高效液相色谱法,测定时间比普通的高效液相色谱法缩短了约40 min. 结论: 该方法经方法学验证,适用于稳心颗粒中皂苷类成分的快速测定.

Objective: To develop a novel method for the rapid extraction and determination of notoginsenoside R₁,ginsenoside Rg₁ and ginsenoside Rb₁ in Wenxin granules by accelerated solvent extraction(ASE)coupled with ultra-performance liquid chromatography(UPLC). Methods: Ether was used to remove the lipid ingredients and water-saturated butanol solution was adopted to extract the saponins.The analysis was performed on an Agilent Eclipse Pluse C₁₈ column(2.1 mm×50 mm,1.8 μm),the mobile phase consisted of acetonitrile(A)and water(B)with gradient program(0-10 min,81%B;10-17 min,81%B→45%B;17-18 min,45%B→81%B;18-25 min,81%B).The detection wavelength was 203 nm and the column temperature was set at 25℃.In comparison with traditional method,ASE reduced the extraction time from 14 h to 25 min and substantially increased processing efficiency. Results: The linearity was achieved in the range of 0.084-0.84 μg(r=0.999 9)for notoginsenoside R₁,0.201 6-2.016 μg(r=0.999 8)for ginsenoside Rg₁ and 0.208 4-2.084 μg(r=0.999 9)for ginsenoside Rb₁.The average recoveries(n=6)for each were 98.4%(RSD=1.1%),98.2%(RSD=0.47%)and 97.9%(RSD=1.3%).The content of saponins was 2.65 mg·g^-1 essentially in agreement with Chinese pharmacopoeia method(2.63 mg·g^-1).The determination time was 40 minutes shorter by UPLC than by HPLC Conclusion: The method was suitable for the determination of saponins in Wenxin granules.