HPLC法测定蒿秦化斑颗粒中四种成分的含量

Content determination of four components in Haoqin Huaban Granules by HPLC

目的建立蒿秦化斑颗粒中马钱苷酸、龙胆苦苷、芍药苷和5-O-甲基维斯阿米醇苷含量的高效液相色谱(HPLC)测定方法。方法采用Kinetex EVO C18色谱柱(150 mm×4.6 mm,5μm),以乙腈-0.1%磷酸为流动相,梯度洗脱,流速1.0 mL/min,检测波长235 nm,柱温35℃。结果马钱苷酸、龙胆苦苷、芍药苷和5-O-甲基维斯阿米醇苷的标准曲线分别为Y=1.5×10^3X-0.88(r=0.9999)、Y=9.8×10^2X-2.95(r=0.9999)、Y=1.4×10^3X-3.97(r=0.9999)、Y=2.4×10^3X-0.33(r=0.9999);线性范围分别为0.03~0.69 mg/L、0.15~3.00 mg/L、0.17~3.44 mg/L、0.01~0.11 mg/L,平均加样回收率分别为97.85%、98.30%、94.66%、96.08%,RSD分别为1.00%、0.69%、1.72%、1.90%。结论该方法简便、准确、重复性好、专属性强,可用于蒿秦化斑颗粒中四种成分的含量测定。

Objective To establish high-performance liquid chromatography (HPLC) method for the content determination of the loganic acid, gentiopicroside, paeoniflorin and 5-O-methylvisamminol in Haoqin Huaban Granules. Methods The column was Kinetex EVO C18 (150 mm × 4.6 mm, 5 μm). The mobile phase consisted of acetonitrile-0.1% phosphoric acid solution with gradient elution at the flow rate of 1.0 mL/min. The detecting wavelength was 235 nm and the column temperature was 35℃. Results The standard curve of loganic acid, gentiopicroside, paeoniflorin and 5-O-methylvisamminol were Y = 1.5×10^3X - 0.88 (r = 0.9999),Y = 9.8×10^2X - 2.95 (r = 0.9999),Y =1.4×10^3X - 3.97 (r = 0.9999),Y =2.4×10^3X - 0.33 (r = 0.9999), and the linear ranges of loganic acid, gentiopicroside, paeoniflorin and 5-O-methylvisamminol were 0.03-0.69 mg/L, 0.15-3.00 mg/L, 0.17-3.44 mg/L, 0.01-0.11 mg/L,respectively. The average recoveries of loganic acid, gentiopicroside paeoniflorin and 5-O-methylvisamminol were 97.85%, 98.30%, 94.66%, 96.08%, the RSD were 1.00%,0.69%,1.72%,1.90%, respectively. Conclusion The method is simple, accurate and reproducible. It is effective in controlling the quality of Haoqin Huaban Granules.