快速筛查消渴类中成药中非法添加成分的HPLC-DAD数据库的建立

Development of HPLC-DAD Database for Rapid in Situ Screening of Ingredients Illegally Adulterated in Anti-Diabetic Traditional Chinese Medicines

关键词:  消渴类中成药  DAD光谱库  现场分析  快速筛查

Key words:  anti-diabetic traditional Chinese medicines  DAD spectrum library  in situ analysis  rapid screening

摘要:

目的 在等度洗脱条件下建立10种化学降糖成分的HPLC-DAD数据库，用于消渴类中成药中非法添加成分的现场快速筛查。方法 采用Waters XTerra MSC₁₈(4.6 mm×150 mm，5 mm)色谱柱，使用2种流动相：流动相I为乙腈-0.02%磷酸(55︰45)，流动相II为乙腈-0.01%三氟乙酸(25︰75)，检测波长229 nm，参比波长550 nm，波长扫描范围为190~600 nm。首先采集对照品的保留时间和光谱图，建立数据库；然后将样品中检出峰的保留时间与数据库比较，初筛出可疑的化学成分；最后将样品中可疑色谱峰的DAD光谱图与数据库中相应对照品的DAD光谱图比较，快速筛查出非法添加的化学成分，并对检出的化学成分进行LC-MS验证。结果 10种化学降糖成分在上述2种等度洗脱条件下实现了完全分离；采用本试验建立的方法检测了市售12批不同的消渴类中成药，发现其中5批含有非法添加成分，并且有1批同时含有4种非法降糖成分添加剂，LC-MS验证结果准确。结论 本法可以准确有效地快速分离10种化学降糖成分，为消渴类中成药的现场打假提供有力的技术支持。

Abstract:

OBJECTIVE　To develop the HPLC-DAD database of ten kinds of anti-diabetic chemicals at the condition of composition-constant mobile-phase for the rapid and in situ screening of ingredients illegally adulterated in anti-diabetic traditional Chinese medicines. METHODS　HPLC was performed on Waters XTerra MSC₁₈(4.6 mm×150 mm, 5 μm) column with two kinds of composition-constant mobile-phases: mobile phase I, acetonitrile-0.02% phosphoric acid(55︰45); and mobile phase II, acetonitrile-0.01% trifluactic acid (25︰75). The detection was performed at 229 nm with the reference wavelength of 550 nm, and DAD scanning range was 190-600 nm. At first, collect peak retention time and DAD spectrum of each reference for establishing a data library. Then, compare the peak retention time of a sample with that of the references in the database for screening suspicious substances in the sample. Finally, compare the obtained DAD spectrum of the substance with that of references in the DAD spectrum library for the further screening. Confirmation of the detected chemicals was performed by LC-MS. RESULTS　Ten kinds of anti-diabetic medicines were well separated at the above two composition constant mobile phases. The screening results of 12 batches of different anti-diabetic traditional Chinese medicine indicated that there were 5 batches containing illegally adulterated components. And four kinds of illegally added chemicals were detected in 1 batch of samples. The assay results were confirmed by HPLC-MS. CONCLUSION The assay developed here is accurate and rapid in the separation of ten kinds of anti-diabetic chemicals, and can give a strong support for the in situ screening of chemicals illegally added in anti-diabetic traditional Chinese medicine.