亲水作用色谱法测定阿卡波糖含量

Determination of acarbose content by hydrophilic interaction liquid chromatography

目的:建立采用亲水作用色谱(HILIC)测定阿卡波糖含量的新方法。方法:采用亚乙基桥杂化酰胺色谱柱(XBridge Amide,4.6 mm×250 mm,3.5μm),通过正交设计试验考察了流动相pH、盐浓度、柱温及流速等色谱参数的影响,最终确立色谱条件:以10 mmol·L^(-1)乙酸铵缓冲盐水溶液(pH 9.0)-乙腈(25∶75)为流动相,流速1.2 mL·min^(-1),柱温65℃,检测波长210 nm。结果:阿卡波糖与杂质A可完全分离,且阿卡波糖质量浓度在0.2~1.2 mg·mL^(-1)范围内线性关系良好(r=0.999 8);平均回收率为99.5%;供试品溶液在24 h内稳定;检测限(S/N=3)为0.08μg;3批胶囊样品中阿卡波糖含量分别为98.6%、100.0%、98.5%。结论:本法可用于阿卡波糖的质量控制。

Objective. To establish a new method for the determination of the content of acarbose using hydrophilic interaction chromatography （ HILIC ）. Methods ： An ethylenically bridged amide column （ XBridge Amide, 4.6 mm× 250 mm, 3.5 μm ） was adopted. The chromatographic parameters, such as pH value and salt concentration of mobile phase, column temperature and flow rate were optimized through orthogonal design test. The new chromatographic conditions were proposed as follows： The sample was separated on the XBridge Amide column using 10 mmol · L^-1 ammonium acetate buffer solution （ pH 9.0 ） and acetonitrile （ 25 ： 75 ） as the mobile phase at a flow rate of 1.2 mL· min^-1. The column temperature was 65 ℃ and the UV detection wavelength was set at 210 nm. Results： Acarbose can be completely separated with impurity A using this method. The calibration curve was linear in the concentration range of 0.2-1.2 mg· mL-1 with the correlation coefficient （ r ） of 0. 999 8. The average recovery was 99.5% and the sample solution was stable in 24 h. The limit of detection （ S/N=3 ） was 0.08 μg. Acarbose content of three batches of capsule samples was 98.6%, 100.0% and 98.5%, respectively. Conclusion：The results indicate that the established method can be applied to quality control of acarbose.