HPLC-ELSD同时测定丹参川芎嗪注射液中间体中7种糖类成分

Simultaneous Determination of Seven Saccharides in the Intermediates of Danshen Chuanxiongqin Injection by HPLC-ELSD

关键词:  丹参川芎嗪注射液  HPLC-ELSD  糖类

Key words:  Danshen Chuanxiongqin injection  HPLC-ELSD  saccharide

摘要:

目的 建立同时测定丹参川芎嗪注射液中间体中果糖、葡萄糖、蔗糖、蜜二糖、棉子糖、甘露三糖和水苏糖7种糖类成分的HPLC-ELSD方法，并研究丹参川芎嗪注射液生产过程中糖类成分的变化规律。方法 采用Prevail Carbohydrate ES色谱柱（250 mm×4.6 mm，5 μm），以乙腈-水为流动相进行梯度洗脱，柱温为25 ℃，进样量为5 μL，流速为0.6 mL·min^–1，ELSD检测器雾化器温度为70 ℃，漂移管温度为60 ℃，氮气流量1.0 L·min^–1。结果 7种糖类成分在定量范围内线性关系良好，R²均>0.998 0，进样、日内、日间和中间精密度RSD值均<5%，供试品溶液在10 h内稳定，低、中、高浓度平均加样回收率为97.00%~104.34%。结论 建立的分析方法稳定、准确、重复性好，可用于丹参川芎嗪注射液中间体中7种糖类成分含量测定。本研究推进了丹参川芎嗪注射液安全性再评价质量控制研究，也为其他中药制剂中糖类成分的定量、转移、转化规律研究提供了参考。

Abstract:

OBJECTIVE To establish a HPLC-ELSD method for simultaneous determination of seven saccharide components in the intermediates of Danshen Chuanxiongqin injection, including fructose, glucose, sucrose, melibiose, raffinose, manninotriose and stachyose. To study the transfer regularity of saccharides in the manufacturing process of Danshen Chuanxiongqin injection. METHODS Chromatographic column was Prevail Carbohydrate ES column(250 mm×4.6 mm, 5 μm) in the gradient elute mode. The mobile phase was acetonitrile and water. The column temperature was set at 25 ℃, the injection volume was 5 μL and the flow rate was 0.6 mL·min^–1. ELSD vaporizer temperature was set at 70 ℃, the drift tube temperature was set at 60 ℃ and the nitrogen gas flow was 1.0 L·min^–1. RESULTS Seven saccharide components showed good linear relationship(R²>0.998 0) within the quantitative ranges. The RSDs of the injection, intra-day, inter-day and intermediate precision results were all <5%. The test solution was stable within 10 h. The average recoveries at three levels ranged was 97.00%-104.34%. CONCLUSION The stability, repeatability, accuracy of the established method were well validated and proved to be. The method can be used for simultaneous determination of seven saccharide components in the intermediates of Danshen Chuanxiongqin injection. This study promotes the safety reevaluation and quality control of Danshen Chuanxiongqin injection and provides a direction for the quantification and transformation researches of saccharide components in other traditional Chinese medicine preparations.