黄芪药材中黄芪甲苷UPLC-ELSD含量测定方法的优化

Optimization of Determination of Astragaloside IV in Astragali Radix

摘要:

目的: 优化黄芪甲苷含量测定的供试液制备方法,建立黄芪药材中黄芪甲苷UPLC-ELSD含量测定的方法。方法: 采用UPLC-ELSD测定黄芪甲苷含量流动相乙腈-水(32:68),柱温30 ℃,流速0.5 mL·min^-1,ELSD检测器温度45 ℃,载气干燥压缩空气,压力0.5 kPa,增益值7。采用甲醇和氨水混合超声提取黄芪中黄芪甲苷,经正丁醇萃取,通过单因素试验优选供试品溶液的制备方法。结果: 供试品制备方法为加甲醇80 mL和浓氨水40 mL,超声提取30 min,用水饱和正丁醇萃取4次,每次40 mL。采用优化的方法测得11批样品中黄芪甲苷含量明显高于2010年版《中国药典》黄芪项下含测方法的结果。结论: 优化的供试液制备方法操作简单、有效成分损失少。建立的含量测定方法准确可靠、灵敏度高。

Abstract:

Objective: To optimize sample solution preparation method and establish assay method of astragaloside IV in Astragali Radix. Method: UPLC-ELSD was used to determine the content of astragaloside IV with mobile phase of acetonitrile-water(32: 68),column temperature of 30 ℃,flow rate of 0.5 mL·min^-1,temperature of ELSD at 45 ℃,carrier gas was dry compressed air with pressure of 0.5 kPa,gain value was 7.Ammonia-methanol mixture was used to ultrasonic extract of astragaloside IV as well as n-butanol extraction.Preparation method of test solution was optimized through single-factor tests. Result: Optimal preparation method was as following:added methanol 80 mL and concentrated ammonia 40 mL,ultrasonic extraction for 30 min,purification by subsequent extraction with n-butanol for four times every 40 mL in sample preparation.The content of astragaloside Ⅳ from 11 batches of Astragali Radix by this optimizing method were significantly higher than the result determined by HPLC method in 2010 edition of Chinese Pharmacopoeia. Conclusion: This optimized sample preparation method is simple with less loss of effective constituents.UPLC analysis method is accurate and reliable with high sensitivity.