HPLC-DAD数据库快速筛查减肥类中药制剂及保健食品中的10种非法添加化学药物

A HPLC-DAD spectrum library for rapidly detecting 10 anti-obesity chemicals illegally adulterated in traditional Chinese medicines and healthy foods

目的:在等度洗脱条件下建立10种有减肥作用化学药物的HPLC-DAD分析方法及数据库,用于减肥类中药制剂及保健食品中非法添加物的快速鉴别。方法:采用Alltina C18色谱柱,使用两种等组成流动相:Ⅰ为乙腈-0.02 mol.L-1乙酸铵(含0.1%乙酸)(12∶88),Ⅱ为乙腈-0.02 mol.L-1乙酸铵(含0.1%乙酸)(60∶40)。检测波长215 nm,参比波长550 nm,波长扫描范围190～600 nm。首先采集对照品的保留时间和光谱图,建立标准谱库;然后将样品中各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种有减肥作用的化学药物在等度洗脱条件下实现了完全分离;并通过数据库快速筛查了1种减肥类中药制剂和8种减肥类保健食品,其中1种减肥保健食品中检出二甲双胍及芬氟拉明,2种减肥保健食品中检出西布曲明及酚酞,其余5种保健食品中检出西布曲明,LC-MS验证结果准确。结论:采用建立的HPLC-DAD光谱库可快速筛选减肥类样品,适合现场采用车载或便携式高效液相色谱仪进行快速筛查的需要;本实验建立的色谱条件可以用于快速筛查减肥类保健食品中非法添加的化学成分。

Objective： To develop the HPLC-DAD spectrum library and analytic procedure at the condition of composition-constant mobile-phase for rapid detection of 10 anti-obesity chemical drugs illegally adulterated in traditional Chinese medicines and healthy foods.Methods：The separation process was performed on an Alltina C18（150 mm×4.6 mm,5 μm） column by two composition-constant mobile-phases with different polarity;i.e.mobile phase I,acetonitrile-0.02 mol·L-1 ammonium acetate（containing 0.1% acetic acid,12∶88）;mobile phase II,acetonitrile-0.02 mol·L-1 ammonium acetate（containing 0.1% acetic acid,60∶40）.The detection wavelength and the reference wavelength were 215 and 550 nm,respectively;the scanning range of DAD was 190~600 nm.Firstly,DAD spectrum library was established by collecting the data of peak retention time and DAD spectrum of the references;secondly,chromatogram comparison between the peak retention time of a sample with that of references,preliminary screening of the suspicious substances in the sample;finally,comparing the spectrum of suspicious substance with the corresponding reference spectrum in DAD spectrum library.The result was confirmed by HPLC-MS.Results： Ten anti-obesity chemicals were well separated by composition-constant mobile-phase.One of traditional Chinese medicines and eight kinds of healthy foods were rapidly detected by the proposed method.The detected metformin and fenfluramine were illegally adulterated in one healthy food;sibutramine and phenolphthalein were illegally adulterated in two kinds of healthy foods;sibutramine was illegally adulterated in the other five healthy foods.The results were further justified by LC-MS method.Conclusion： DAD spectrum library is usable for rapid point-of-care test of unknown chemicals.The HPLC-DAD conditions developed in our method are suitable for the rapid screening of chemicals in anti-obesity traditional Chinese medicines and healthy foods.