HPLC法同时测定肝炎康复丸中6种成分的含量

Simultaneous Determination of Six Components in Ganyan Kangfu Pills by HPLC

目的:建立HPLC波长切换联合梯度洗脱法同时测定肝炎康复丸中6种主要成分[(R,S)-告依春、丹酚酸B、丹参酮Ⅱ_A、木犀草苷、对羟基苯乙酮、滨蒿内酯]的含量。方法:色谱柱为Agilent TC-C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液梯度洗脱,流速为0.9 ml·min^(-1),检测波长分别为245 nm[检测(R,S)-告依春]、270 nm(检测丹酚酸B和丹参酮Ⅱ_A)、348 nm(检测木犀草苷)、278 nm(检测对羟基苯乙酮和滨蒿内酯),柱温为25℃,进样量为10μl。结果:(R,S)-告依春、丹酚酸B、丹参酮Ⅱ_A、木犀草苷、对羟基苯乙酮、滨蒿内酯的线性范围分别为1.99~49.75μg·ml^(-1)(r=0.999 9)、18.66~466.50μg·ml^(-1)(r=0.999 4)、2.25~56.25μg·ml^(-1)(r=0.999 8)、2.62~65.50μg·ml^(-1)(r=0.999 8)、2.48~62.00μg·ml^(-1)(r=0.999 2)和2.55~63.75μg·ml^(-1)(r=0.999 6),平均加样回收率分别为98.42%、99.56%、97.96%、96.84%、98.10%和97.82%,RSD分别为0.83%、1.04%、1.53%、0.78%、1.44%和1.34%(n=6)。结论:本方法简便、准确、重复性好,可为肝炎康复丸多指标定量质量评价提供依据。

Objective： To establish a wavelength switching HPLC with gradient elution for the simultaneous determination of six active components, [ （ R, S） -goitrin, salvianolic acid B, tanshinone IIA, luteolin-7-glucoside, 4-hydroxyacetophenone and scoparone ] in Ganyan Kangfu pills. Methods： An Agilent TC-C18 （250 mm ×4.6mm,5μm） chromatographic column was used with the mobile phase consisting of acetonitrile and 0.1% phosphoric acid solution with gradient elution. The flow rate was 0.9 ml min-1 , and the column temperature was 25 ℃. （R,S）-Goitrin was detected at 245 nm, salvianolic acid B and tanshinone IIA were detected at 270 nm, lnteolin-7-glucoside was detected at 348 nm, 4-hydroxyacetophenone and scoparone were detected at 278 nm. The sample volume was 10 p,1. Results： The linear range of （ R, S） -goitrin, salvianolic acid B, tanshinone ⅡA, luteolin-7-glucoside, 4-hydroxyacetophe- none and scoparone was 1.99-49.75 μg·ml-1（ r = 0.999 9 ）, 18.66-466.50 μg·ml-1 （ r = 0.999 4 ）, 2.25-56.25 μg·ml-1（ r = 0.999 8）, 2.62-65.50 μg·ml-1 （r =0.999 8）, 2.48-62.00μg·ml-1 （r =0.999 2） and 2.55-63.75μg·ml-1 （r =0.999 6）, respectively. The average recovery was 98.42% （RSD =0.83% ）, 99.56% （RSD = 1.04% ）, 97.96% （RSD = 1.53% ）, 96.84% （RSD = 0.78% ）, 98.10% （ RSD = 1.44% ） and 97.82% （ RSD = 1.34% ） （ n = 6 ）, respectively. Conclusion： The method with good reproducibility is simple and accurate, which provides a new way for the quality evaluation of Ganyan Kangfu pills.