目的 建立虎驹乙肝胶囊的HPLC-ELSD和HPLC-UV 2种特征图谱以及胶囊中虎杖苷、丹酚酸B和五味子醇甲3种活性成分的定量测定方法.方法 HPLC-ELSD特征图谱:Thermo C₁₈(250 mm×4.6 mm,5 μm)色谱柱,以乙腈-水为流动相梯度洗脱,漂移管温度85 ℃,气体压力275.80 kPa,增益500,柱温35 ℃,体积流量1 mL/min,进样量10 μL.HPLC-UV特征图谱:Hedra ODS-2(250 mm×4.6 mm,5 μm)色谱柱,流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,体积流量为0.8 mL/min,检测波长为215 nm,柱温30 ℃,进样量10 μL.结果 建立了虎驹乙肝胶囊的HPLC-ELSD和HPLC-UV 2个特征图谱,分别确定了10、12个共有峰,指认其中的11个共有峰为虎杖苷、三七皂苷R₁、人参皂苷Rg₁、人参皂苷Rb₁、黄芪甲苷、绿原酸、大黄素、丹酚酸B、五味子醇甲、五味子甲素和五味子乙素,共有峰的参数符合"中药注射剂指纹图谱技术要求"的有关规定.定量测定中虎杖苷、丹酚酸B、五味子醇甲3种活性成分的分离度良好,线性范围分别为2.24～287.33 μg/mL(r=0.999 9)、1.95～250 μg/mL(r=0.999 9)和1.10～141.66 μg/mL(r=0.999 9),测定结果具有很好的重复性和稳定性.结论 特征图谱结合3种成分的定量测定,能更好地控制虎驹乙肝胶囊的质量.

Objective To establish the HPLC-ELSD and HPLC-UV specific chromatograms of Huju Yigan Capsule and an HPLC method for simultaneous determination of three active ingredients. Methods HPLC-ELSD: HPLC analysis was performed on a Thermo C₁₈ (250 mm × 4.6 mm, 5 μm) column with a mixture of acetonitrile and water as mobile phase in gradient mode. The drift tube temperature was at 85 ℃, gas pressure was 275.80 kPa, gain value was 500, and column temperature was at 35 ℃. HPLC-UV: The Hedra OSD-2 (250 mm × 4.6 mm, 5 μm) column was used. Acetonitrile and water containing 0.1% phosphate were used as mobile phase of HPLC with gradient elution, monitored with the wavelength of 215 nm. Flow rate was set at 0.8 mL/min and column temperature was 30 ℃. Results The two chromatographic fingerprints were completed with 10 and 12 recognizable peaks, of which 11 peaks (polydatin, notoginsenosides R₁, ginsenosides Rg₁, Rb₁, astragaloside, chlorogenic acid, emodin, bsalvianolic acid B, schisandrin, schizandrin A, and γ-schizandrin B) were determined. The parameters of the common peaks were accorded with the stipulation of "Traditional Chinese Medicinal Injection Fingerprint Technical Requirements". Three active components were finely separated and showed good linearity in the concentration range of detection with repeatability and stability, and the ranges were 2.24—287.33 μg/mL for polydatin (r = 0.999 9), 1.95—250 μg/mL for bsalvianolic acid B (r = 0.999 9), and 1.10—141.66 μg/mL for schisandrin (r= 0.999 9). Conclusion The combination of specific chromatograms and determination of three components is capable of effectively controlling of the quality of Huju Yigan Capsule.

江苏高校优势学科建设工程资助项目(PAPD);江苏省"青蓝工程"创新团队支持计划资助(QLP)