基于HPLC-DAD波长切换法同时测定祛风止痛胶囊中5种成分的含量

Simultaneous Determination of the Contents of Five Components in Qufeng Zhitong Capsules(祛风止痛胶囊)Based on HPLC-DAD Wavelength Switching Method

目的:建立祛风止痛胶囊中羟基红花黄色素A、山柰素、川续断皂苷Ⅵ、没食子酸、原儿茶酸5种成分的含量测定方法。方法:采用HPLC波长切换法同时测定祛风止痛胶囊中羟基红花黄色素A、山柰素、川续断皂苷Ⅵ、没食子酸、原儿茶酸5种化学成分的含量,色谱柱为Waters XBridge TM C18柱(250 mm×4.6 mm,5μm);以甲醇-0.5%磷酸水溶液为流动相,梯度洗脱;柱温为30℃;检测波长分别为λ_(1)=212 nm、λ_(2)=280 nm、λ_(3)=367 nm。结果:羟基红花黄色素A、山柰素、川续断皂苷Ⅵ、没食子酸、原儿茶酸分离度良好;分别在0.809~16.170μg/mL(r=0.999 8)、3.469~69.380μg/mL(r=0.999 9)、3.420~68.400μg/mL(r=0.999 7)、0.907~18.130μg/mL(r=0.999 9)、1.250~24.990μg/mL(r=0.999 8)的范围内线性关系良好;平均加样回收率分别为99.99%(RSD=0.65%)、102.43%(RSD=2.48%)、103.88%(RSD=1.54%)、99.85%(RSD=0.44%)、103.40%(RSD=0.73%)。结论:本研究建立了同时测定祛风止痛胶囊中5种指标性成分的含量方法,该法操作准确、重复性好,可客观、全面地对祛风止痛胶囊的内在质量进行有效评价。

Objective: To establish a method for the determination of five components of hydroxysafflor yellow A, Kaempferide, AsperosaponinⅥ, gallic acid and protocatechuic acid in Qufeng Zhitong Capsules. Methods: The contents of hydroxysafflor yellow A, kaempferide, asperosaponin VI, gallic acid and protocatechuic acid in the compound were detected by HPLC-ELSD with wavelength switching combined gradient elution method with a Waters XBridgeTMC18 column(4.6 mm×250 mm, 5 μm), a mobile phase was acetonitrile-0.1% phosphoric acid with, velocity of flow was 1.0 mL/min. The temperature of column was 30 ℃, and detection wavelength was set atλ_(1)=212 nm, λ_(2)=280 nm, λ_(3)=367 nm. Results: Hydroxysafflor yellow A, kaempferide, asperosaponin VI, gallic acid and protocatechuic acid displayed good linear relationships within the range of 0.809-16.170 μg/mL(r=0.999 8),3.469-69.380 μg/mL(r=0.999 9), 3.420-68.400 μg/mL(r=0.999 7), 0.907-18.130 μg/mL(r=0.999 9), 1.250-24.990μg/mL(r =0.999 8). The average recoveries were 99.99%(RSD =0.65%), 102.43%(RSD =2.48%), 103.88%(RSD =1.54%), 99.85%(RSD=0.44%), 103.40%(RSD=0.73%), respectively. Conclusion: This study established a method for the simultaneous determination of five components in Qufeng Zhitong Capsules. The method is accurate and reproducible, and can objectively and comprehensively evaluate the internal quality of Qufeng Zhitong Capsules.