基于高效液相色谱波长切换技术评价活血止痛散中当归和乳香质量

Quality evaluation of Angelicae Sinensis Radix and Olibanum in Huoxue Zhitong powders based on HPLC combined with wavelength switching

目的:采用HPLC法结合波长切换技术建立活血止痛散中当归和乳香的质量评价方法。方法:样品经70%乙醇超声提取,测定阿魏酸和11-羰基-β-乙酰乳香酸含量,同时检查松香酸。采用Agilent TC-C_(18)色谱柱(4.6 mm×250 mm,5μm)分离,柱温30℃。乙腈-0.1%甲酸梯度洗脱(0～8 min:30%乙腈;8～10 min:30%～80%乙腈;10～30 min:80%乙腈),流速1.0 mL·min^(-1)。321 nm下检测阿魏酸(0～8 min);240 nm下检测松香酸(8～23 min);250 nm下检测11-羰基-β-乙酰乳香酸(23～30 min)。结果:松香酸检出限为3.13 ng·mL^(-1)。阿魏酸和11-羰基-β-乙酰乳香酸分别在0.01～100μg·mL^(-1)和0.2～200μg·mL^(-1)范围内线性关系良好,平均回收率分别为100.3%和99.7%。3个厂家的7批样品阿魏酸含量在0.19～0.37 mg·g^(-1)之间,11-羰基-β-乙酰乳香酸含量在2.36～3.48 mg·g^(-1)之间。结论:所建方法准确、简便、快速,可为活血止痛散的全面质量控制提供参考。

OBJECTIVE To establish the evaluation method for quality of Angelicae Sinensis Radix and Olibanum in Huoxue Zhitong powders by high performance liquid chromatography(HPLC)method combined with wavelength switching detection technology.METHODS The samples were extracted with 70%ethanol by ultrasonic to determine the contents of ferulic acid and 11-carbonyl-β-acetyl boswellic acid.The separation was performed on an Agilent TC-C18(4.6 mm×250 mm,5μm)column with a column temperature of 30℃.The mobile phase was composed of acetonitrile and 0.1%formic acid with gradient elution(0-8 min:30%acetonitrile;8-10 min:30%-80%acetonitrile;10-30 min:80%acetonitrile)at a flow rate of 1.0 mL·min-1.Ferulic acid was detected at 321 nm(0-8 min);abietic acid was detected at 240 nm(8-23 min);acetyl-11-keto-β-boswellic acid was detected at 250 nm(23-30 min).RESULTS The limit of detection of abietic acid was 3.13 ng·mL-1.The calibration curves of ferulic acid and acetyl-11-keto-β-boswellic acid showed good linearity within the range of 0.01-100μg·mL-1 and 0.2-200μg·mL-1,respectively.And the average recoveries for the above analytes were 100.3%and 99.7%,respectively.The contents of ferulic acid and acetyl-11-keto-β-boswellic acid in 7 batches of samples from 3 manufactures were within 0.19-0.37 mg·g-1 and 2.36-3.48 mg·g-1,respectively.CONCLUSION The proposed method is accurate,simple and rapid,thus providing basis for comprehensive quality control of Huoxue Zhitong powder.