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RP-HPLC法测定活血化瘀方中7种有效成分
陈琳1,张新新1,党旋1,张天龙2,李振志1,赵晔1*
0
(1. 西北大学陕西省生物医药重点实验室,西安 710069
2. 西北大学生命科学学院中药学系,西安 710069
*通信作者)
摘要:
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 ShieldTM RP C18(150 mm×4.6 mm, 3.5 μm)色谱柱,以甲醇-0.25%(体积比)冰醋酸水溶液为流动相,流速为0.8 mL/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280 nm;黄芪甲苷采用蒸发光散射检测器,飘移管温度90℃,气流量2.8 L/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0.01~0.80 μg(r=0.999 8),0.005~0.4 μg(r=0.999 7),0.05~4 μg (r=0.999 8),0.005~0.4 μg(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);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、3.12 mg/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。
关键词:  活血化瘀方;丹参素;原儿茶醛;芍药苷;葛根素;阿魏酸;丹参酮ⅡA;黄芪甲苷  高压液相色谱法
DOI:
投稿时间:2013-05-28修订日期:2013-08-08
基金项目:西北大学 “十二五”“211工程”研究生创新人才培养项目(YZZ12059).
RP-HPLC in determination of seven active ingredients in blood-invigorating and stasis-removing prescription
CHEN Lin1,ZHANG Xin-xin1,DANG Xuan1,ZHANG Tian-long2,LI Zhen-zhi1,ZHAO Ye1*
(1. Biomedicine Key Laboratory of Shaanxi Province, Northwest University, Xi’an 710069, Shaanxi, China
2. Department of Traditional Chinese Medicine, College of Life Sciences, Northwest University, Xi’an 710069, Shaanxi, China
*Corresponding author.)
Abstract:
Objective To develop an RP-HPLC method for determination of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone ⅡA and astragaloside in blood-invigorating and stasis-removing prescription (BSP). Methods The analysis was performed with a column of Waters Symmetry ShieldTM RP C18 (150 mm × 4.6 mm, 3.5 μm), and the mobile phase consisted of methanol-0.25% acetic acid. The flow rate was 0.8 mL/min and the column temperature was 30℃. UV was employed to determine the contents of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid and tanshinone ⅡA, and the detection wavelength was set at 280 nm. Evaporative light scattering detection (ELSD) was employed to determine the contents of astragaloside. The temperature of drift tube was 90℃ and the gas flow was 2.8 L/min (compressed air). Results The linearity was obtained over 0.01-0.80 μg (r=0.999 8) for tanshinol, 0.005-0.4 μg (r=0.999 7) for protocatechualdehyde, 0.05-4 μg (r=0.999 8) for paeoniflorin, 0.005-0.4 μg (r=0.999 7) for puerarin, 0.006-0.5 μg (r=1) for ferulic acid, 0.005-0.4 μg (r=1) for tanshinone ⅡA, and 0.031-2.46 μg (r=0.999 3) for astragaloside. The recoveries were all between 97.0%-101.0%, and RSDs were all less than 2%. The contents (mean) of tanshinol, protocatechualdehyde, paeoniflorin, puerarin, ferulic acid, tanshinone ⅡA and astragaloside in three batches of samples were 1.15, 0.13, 4.48, 0.80, 0.72, 0.31 and 3.12 mg/g, respectively. Conclusion The method in our study is convenient, accurate and sensitive, and it provides a reference for the determination of active ingredients in BSP.
Key words:  blood-invigorating and stasis-removing prescription  tanshinol  protocatechualdehyde  paeoniflorin  puerarin  ferulic acid  tanshinone ⅡA  astragaloside  high pressure liquid chromatography