高效液相色谱法测定川续断中熊果酸、齐墩果酸、马钱苷和川续断皂苷Ⅵ的含量

目的:利用高效液相色谱法(HPLC)建立川续断中熊果酸、齐墩果酸、马钱苷和川续断皂苷Ⅵ4种指标成分的含量测定方法。方法:(1)测定熊果酸、齐墩果酸色谱条件为:色谱柱:Agilent ZORBAX SB-C₁₈(4.6 mm×250 mm,5 μm);流动相:甲醇-0.2%醋酸铵水溶液(86:14);流速:0.9 ml·min^-1;柱温:25 ℃;检测波长:210 nm。(2)测定马钱苷、川续断皂苷Ⅵ色谱条件为:色谱柱:Agilent ZORBAX SB-C₁₈(4.6 mm×250 mm,5 μm);流动相:0~8 min乙腈-水(15:85),8~10 min乙腈-水(25:75),10~28 min乙腈-水(30:70);流速:0.8 ml·min^-1;柱温:25 ℃;分别在240 nm、210 nm的波长条件下检测马钱苷和川续断皂苷Ⅵ。结果:(1)熊果酸在5.20~520 μg·ml^-1范围内线性关系良好,r=0.999 7;平均回收率为96.4%,RSD为0.96%;齐墩果酸在5.64~564 μg·ml^-1范围内线性关系良好,r=0.999 8;平均回收率为97.7%,RSD为1.66%。(2)马钱苷在6.04~604 μg·ml^-1范围内线性关系良好,r=0.999 9;平均回收率为99.4%, RSD为1.35%,川续断皂苷Ⅵ 6.28~628 μg·ml^-1范围内线性关系良好,r=1.000;平均回收率为97.7%,RSD为0.90%。结论:该方法简单、准确可行,结果稳定可靠,可为川续断的质量控制提供科学依据。

Contents of ursolic acid,oleanolic acid,loganin and akebia saponin D in Dipsacus asper determined by HPLC

OBJECTIVE To establish the method to measure contents of ursolic acid, oleanolic acid, loganin and akebia saponin D in Dipsacus asper by high performance liquid chromatography (HPLC). METHODS Chromatographic conditions for content determination of ursolic acid and oleanolic acid: column: Agilent ZORBAX SB-C₁₈(4.6 mm×250 mm, 5 μm); mobile phase: methanol-2% ammonium acetate aqueous solution (86:14); flow velocity: 0.9 ml·min^-1; column temperature: 25 ℃; detection wavelength: 210 nm. Chromatographic conditions for content determination of loganin and akebia saponin D: column: Agilent ZORBAX SB-C₁₈(4.6 mm×250 mm, 5 μm); mobile phase: 0-8 min acetonitrile-water (15:85), 8-10 min acetonitrile-water (25:75), 10-28 min acetonitrile-water (30:70); flow velocity: 0.8 ml·min^-1; column temperature: 25 ℃; detection wavelength: 240 nm and 210 nm.RESULTS Good linearity within the range of 5.20-520 μg·ml^-1 for ursolic acid, 5.64-564 μg·ml^-1 for oleanolic acid, 6.04-604 μg·ml^-1 for loganin and 6.28-628 μg·ml^-1 for akebia saponin D. The average recovery of them were 96.4%, 97.6%, 99.4% and 97.7%, respectively. CONCLUSION The method is simple, accurate and feasible. The results are stable and reliable. And can provide a scientific basis for quality control of Dipsacus asper.