目的 建立HPLC法同时测定参乌益肾片中β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷8种成分，联合化学计量学方法对参乌益肾片的产品质量进行综合评价。方法 采用Sepax Bio-C₁₈色谱柱（250 mm×4.6 mm，5 μm）；以乙腈–0.1%甲酸进行梯度洗脱；检测波长分别为250 nm（0～20 min测定β-蜕皮甾酮、25R-牛膝甾酮和25S-牛膝甾酮）、330 nm（20～60 min测定大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷）；柱温30℃；体积流量1.0 mL/min；进样量10 μL。采用聚类分析、主成分分析和偏最小二乘法–判别分析法对8种指标成分检测结果相关性进行分析评价。结果 β-蜕皮甾酮、25R-牛膝甾酮、25S-牛膝甾酮、大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木通苯乙醇苷B和2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷分别在1.47～36.75、0.66～16.50、0.74～18.50、5.80～145.00、15.77～394.25、4.58～114.50、1.29～32.25、8.78～219.50 μg/mL有良好的线性关系；平均回收率分别为97.78%、96.90%、97.96%、99.67%、100.01%、99.54%、98.03%、100.11%，RSD值分别为1.17%、0.82%、1.48%、0.93%、0.77%、0.69%、1.13%、0.65%。成分8（2,3,5,4′-四羟基二苯乙烯-2-O-β-D-葡萄糖苷VIP＝1.503）、成分4（大车前苷AVIP＝1.404）、成分5（毛蕊花糖苷VIP＝1.274）、成分1（β-蜕皮甾酮VIP＝1.030）是影响参乌益肾片样品质量的关键成分。结论 建立的方法简便快捷、准确可靠，可用于参乌益肾片中多指标成分的质量综合评价模式。

Objective To establish an HPLC method for simultaneous determination of β-ecdysterone, 25R-inokosterone, 25S- inokosterone, plantamajoside, verbascoside, isoacteoside, calceolarioside B, and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside, and to evaluate the comprehensive quality of Shenwu Yishen Tablets by chemometrics analysis. Methods The separation was carried out on Sepax Bio-C₁₈ column (250 mm×4.6 mm, 5 μm). The mobile phase consisted of acetonitrile - 0.1% formic acid with gradient elution. The detection wavelengths were set at 250 nm for β-ecdysterone, 25R-inokosterone, and 25S-inokosterone, and 330 nm for plantamajoside, verbascoside, isoacteoside, calceolarioside B, and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside. Temperature of column was set at 30 ℃, the flow rate was 1.0 mL/min, and the volume of injection was 10 μL. The chemometrics method was used to analyze and evaluate the correlation of the detection results of eight index components. Results There were good linear relationships of β-ecdysterone, 25R-inokosterone, 25S-inokosterone, plantamajoside, verbascoside, isoacteoside, calceolarioside B, and 2,3,5,4'- tetrahydroxystilbene-2-O-β-D-glucoside within the ranges of 1.47 — 36.75, 0.66 — 16.50, 0.74 — 18.50, 5.80 — 145.00, 15.77 — 394.25, 4.58 — 114.50, 1.29 — 32.25, and 8.78 — 219.50 μg/mL, whose average recoveries and RSD were 97.78% (1.17%), 96.90% (0.82%), 97.96% (1.48%), 99.67% (0.93%), 100.01% (0.77%), 99.54% (0.69%), 98.03% (1.13%), and 100.11% (0.65%), respectively. Through chemometrics analysis, 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(VIP = 1.503), plantamajoside (VIP = 1.404), verbascoside (VIP = 1.274), and β-ecdysterone (VIP = 1.030) were the differential markers affecting the quality of Shenwuyishen Tablets. Conclusion The method is simple and accurate, which can be used for quality evaluation of multiple index components in Shenwu Yishen Tablets.

R286.02

北京市中医管理局第二批中药骨干人才培养项目(京中医科字[2018]213号)