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目的建立RP-HPLC法测定活血化瘀方(blood-invigorating and stasis-removing prescription,BSP)中丹参素、原儿茶醛、芍药苷、葛根素、阿魏酸、丹参酮ⅡA及黄芪甲苷含量的方法。方法采用Waters Symmetry ShieldTMRP C18(150mm×4.6mm,3.5μm)色谱柱,以甲醇-0.25%(体积比)冰醋酸水溶液为流动相,流速为0.8mL/min,柱温30℃。丹参素、原儿茶醛、芍药苷、葛根素、阿魏酸和丹参酮ⅡA采用紫外检测器,检测波长为280nm;黄芪甲苷采用蒸发光散射检测器,飘移管温度90℃,气流量2.8L/min(压缩空气)。结果丹参素、原儿茶醛、芍药苷、葛根素、阿魏酸、丹参酮ⅡA和黄芪甲苷的线性范围分别为0.01～0.80μg(r=0.999 8),0.005～0.4μg(r=0.999 7),0.05～4μg(r=0.999 8),0.005～0.4μg(r=0.999 7),0.006～0.5μg(r=1),0.005～0.4μg(r=1),0.031～2.46μg(r=0.999 3);加样回收率均在97.0%～101.0%之间,且RSD均小于2%。测定了3批样品中丹参素、原儿茶醛、芍药苷、葛根素、阿魏酸、丹参酮ⅡA和黄芪甲苷的含量(平均值)分别为1.15、0.13、4.48、0.80、0.72、0.31、3.12mg/g。结论该方法简便、准确、灵敏,为测定BSP中的有效成分的含量提供参考。
Objective To establish a RP-HPLC method for the determination of Danshensu, protocatechuic aldehyde, paeoniflorin, puerarin, ferulic acid, tanshinone ⅡA and astragaloside in blood-invigorating and stasis-removing prescription (BSP) method. Methods A Waters Symmetry Shield TMRP C18 (150mm × 4.6mm, 3.5μm) column was used. The mobile phase was methanol - 0.25% (v / v) glacial acetic acid and the flow rate was 0.8mL / min. Danshensu, protocatechuic aldehyde, paeoniflorin, puerarin, ferulic acid and tanshinone Ⅱ A using UV detector, detection wavelength of 280nm; Astragaloside using evaporative light scattering detector, the drift tube temperature of 90 ℃, the flow of 2.8L / min (compressed air). Results The linear ranges of danshensu, protocatechuic aldehyde, paeoniflorin, puerarin, ferulic acid, tanshinone ⅡA and astragaloside were 0.01-0.80μg (r = 0.999 8), 0.005-0.4μg (r = 0.999 7 (R = 0.999 7), 0.006-0.5 μg (r = 1), 0.005-0.4 μg (r = 1), 0.031-2.46 μg (r = 0.999 3). The recoveries of samples were between 97.0% and 101.0% with RSD less than 2%. The contents (average) of Danshensu, protocatechuic aldehyde, paeoniflorin, puerarin, ferulic acid, tanshinone ⅡA and astragaloside in the three batches of samples were determined respectively as 1.15,0.13,4.48,0.80,0.72,0.31, 3.12 mg / g. Conclusion The method is simple, accurate and sensitive to provide a reference for the determination of the content of active ingredients in BSP.