熟三七饮片HPLC指纹图谱的建立及多成分定量测定

HPLC fingerprint of steamed Panax Notoginseng and content determination of multi-components

目的建立不同产地熟三七饮片的HPLC指纹图谱,并同时测定其中11种皂苷类成分含量。方法采用Agilent Zorbax SB-C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈-水为流动相(洗脱程序:0~30 min,20%乙腈;30~60 min,20%~45%乙腈;60~88 min,45%~75%乙腈;88~98 min,75%乙腈;98~100 min,75%~20%乙腈),体积流量1.0 m L/min,检测波长203 nm,柱温35℃,建立熟三七饮片的HPLC指纹图谱,并对三七皂苷R1及人参皂苷Rg1、Re、Rb1、20S-Rh1、20R-Rh1、Rd、Rk3、Rh4、20S-Rg3、20R-Rg3等指标成分的定量测定方法进行方法学考察,对10批(S1~S10)熟三七饮片进行含量测定。结果建立了熟三七饮片的HPLC指纹图谱,共标定30个共有峰,S1~S10指纹图谱的相似度分别为0.941、0.938、0.945、0.951、0.913、0.909、0.920、0.928、0.917、0.919;在所建立的定量测定方法条件下,同时测定11个皂苷成分,三七皂苷R1及人参皂苷Rg1、Re、Rb1、20S-Rh1、20R-Rh1、Rd、Rk3、Rh4、20S-Rg3、20R-Rg3平均质量分数分别为5.274、20.515、2.838、23.651、3.476、1.407、5.239、1.784、1.580、0.904、0.294 mg/g;色谱峰均达到了较好地分离,且在线性范围内线性关系良好,11种皂苷的线性回归方程相关系数分别为0.999 7、0.999 5、0.999 5、0.999 7、0.999 7、0.999 6、0.999 7、0.999 6、0.999 7、0.999 7、0.999 6;平均加样回收率分别为101.23%、98.52%、97.67%、99.62%、98.17%、98.92%、99.44%、99.14%、100.25%、98.23%、96.89%;RSD值分别为1.35%、1.58%、2.44%、1.05%、1.48%、1.56%、0.85%、2.34%、2.85%、1.25%、1.08%。结论该方法灵敏、准确、重复性好,可为熟三七饮片的质量评价提供参考。

Objective To establish the HPLC fingerprint and simultaneous method for the determination of 11 saponins in steamed Panax notoginseng from different origins. Methods Agilent Zorbax SB-C18（250 mm × 4.6 mm, 5 μm） column was adopted, the mobile phase consisted of acetonitrile-water with gradient elution at the flow rate of 1.0 m L/min（elution program： 0—30 min, 20%acetonitrile; 30—60 min, 20%—45% acetonitrile; 60—88 min, 45%—75% acetonitrile; 88—98 min, 75% acetonitrile; 98—100 min, 75%—20% acetonitrile）, and the detection wavelength was 203 nm. The establishment of steamed P. notoginseng HPLC fingerprint, and determination method of notoginsenoside R1, ginsenoside Rg1, Re, Rb1, 20 S-Rh1, 20 R-Rh1, Rd, Rk3, Rh4, 20 S-Rg3, and 20 R-Rg3 index components were studied methodologically. The content of 11 saponins in 10 batches was determined. Results The HPLC fingerprint was establish, and thirty common peaks were selected as characteristic peaks of steamed P. notoginseng. The similarities of different samples from 10 areas were 0.941, 0.938, 0.945, 0.951, 0.913, 0.909, 0.920, 0.928, 0.917, and 0.919. In quantitative analysis, eleven saponins were separated well and the average content was 5.274, 20.515, 2.838, 23.651, 3.476, 1.407, 5.239, 1.784, 1.580, 0.904, and 0.294 mg/g, respectively. Additionally, all calibration curves showed good linear regression relationship, with correlation indexes of 0.999 7, 0.999 5, 0.999 5, 0.999 7, 0.999 7, 0.999 6, 0.999 7, 0.999 6, 0.999 7, 0.999 7, and 0.999 6; The average recoveries were 101.23%, 98.52%, 97.67%, 99.62%, 98.17%, 98.92%, 99.44%, 99.14%, 100.25%, 98.23%, and 96.89%, with RSDs of 1.35%, 1.58%, 2.44%, 1.05%,1.48%, 1.56%, 0.85%, 2.34%, 2.85%, 1.25%, and 1.08%. Conclusion This method is sensitive, accurate and reproducible. It can be used to provide a reference for the standard and evaluation of quality of steamed P. notoginseng.