液相色谱-四极杆飞行时间质谱法对保健食品中30种降糖类非法添加化合物的快速定性检测

Rapid Qualitative Detection of 30 Anti-diabetic Compouds in Health Foods by Liquid Chromatography-Quadrupole/Time-of-Flight Mass Spectrometry

目的:建立液相色谱-四极杆飞行时间质谱法对保健食品中30种降糖类非法添加化合物的快速定性检测方法。方法:样品经甲醇超声提取后,采用Agilent Poroshell 120 EC-C18(150 mm×3.0 mm,2.7μm)色谱柱,10 mmol·L^(-1)甲酸铵的0.1%(v/v)甲酸溶液-乙腈为流动相进行梯度洗脱。30种化合物采用电喷雾离子化正离子检测模式进行分析,建立30种化合物的分子式、一级精确质量、二级碎片离子以及保留时间在内的质谱数据库;在全离子扫描模式下,通过化合物的色谱保留时间、精确质量数、同位素分布和丰度比等信息进行定性匹配;在Target MS/MS模式下,通过二级碎片离子匹配进一步确证,同时以分子离子峰的峰面积定量,实现保健食品中多目标化合物的快速定性定量筛查。结果:该方法能同时实现30种降糖类化学物的快速定性筛查确证,30种化合物在各自的浓度范围内线性良好(r^(2)> 0.99),检出限为5~20 ng·mL^(-1)。在50 ng·mL^(-1)、100 ng·mL^(-1)和200 ng·mL^(-1)的加标水平下,30种化合物在2种基质中的平均回收率在68.7%~110.5%,平均RSD均小于13%。实际样品中筛查出2批添加盐酸二甲双胍。结论:该方法快速、高效、准确、扩展性强,适用于保健食品中降糖类非法添加化合物的高通量快速定性筛查。

Objective:To establish a rapid qualitative detection method for 30 illegally added hypoglycemic compounds in health food using liquid chromatography-quadrupole time-of-flight mass spectrometry.Method:After ultrasonic extraction with methanol,the sample was extracted using Agilent Poroshell 120 EC-C18(150 mm×3.0 mm,2.7μm)chromatographic column was used,with a 0.1%(v/v)formic acid solution of 10 mmol·L^(-1) ammonium formate and acetonitrile as the mobile phase for gradient elution.30 compounds were analyzed by electrospray ionization positive ion detection mode,and a mass spectrometry database including the molecular formula,primary accurate mass,secondary fragment ions and retention time of 30 compounds was established;in the full ion scanning mode,qualitative matching is performed through information such as the chromatographic retention time,accurate mass number,isotope distribution,and abundance ratio of the compound;in Target MS/MS mode,further confirmation is achieved through secondary fragment ion matching,while quantifying the peak area of molecular ion peaks to achieve rapid qualitative and quantitative screening of multi-objective compounds in health food.Result:This method can simultaneously achieve rapid qualitative screening and confirmation of 30 hypoglycemic chemicals,and the 30 compounds have good linearity within their respective concentration ranges(r^(2)>0.99),with a detection limit of 5~20 ng·mL^(-1).At spiked levels of 50 ng·mL^(-1),100 ng·mL^(-1),and 200 ng·mL^(-1),the average recoveries of 30 compounds in two matrices ranged from 68.7%to 110.5%,the average RSD is less than 13%.Two batches of metformin hydrochloride were screened from the actual samples.Conclusion:This method is fast,efficient,accurate,and highly scalable,and is suitable for high-throughput rapid qualitative screening of illegally added hypoglycemic compounds in health food.