超高效液相色谱-三重四极杆质谱法测定保健食品中28种降糖类非法添加化合物

Determination of 28 anti-diabetic compounds in health foods by ultra performance liquid chromatography-triple quadrupole mass spectrometry

中文关键词:  超高效液相色谱-三重四极杆质谱法  非法添加  降糖  保健食品

英文关键词:ultra performance liquid chromatography-tandem mass spectrometry  adulteration  anti-diabetic  health foods

中文摘要:

      目的 建立超高效液相色谱-三重四极杆质谱法测定保健食品中28种降糖类非法添加化合物的分析方法。方法 采用Merck SeQuant ZIC-HILIC色谱柱(100 mm×2.1 mm, 3.5 μm), 以乙腈-含10 mmol/L甲酸铵的0.1%甲酸水溶液为流动相梯度洗脱, 分离二甲双胍等3种化合物; 采用Waters CORTECS T3色谱柱 (100 mm×2.1 mm, 2.7 μm), 以乙腈-0.1%甲酸水溶液为流动相梯度洗脱, 分离苯乙双胍等25种化合物, 在电喷雾离子化正离子或负离子模式下, 以多反应监测方式(multiple reaction monitoring, MRM)检测。结果 二甲双胍等3种化合物在进样浓度为50~1000 μg/L范围内线性关系良好, 妥拉磺脲等5种化合物在25~500 μg/L范围内线性关系良好, 其余20种化合物在5~100 μg/L范围内线性关系良好, 相关系数(r)均大于0.99。各化合物在定量限(limit of quantity, LOQ)1倍、2倍、10倍添加水平下的平均加标回收率为48.6%~134.4%, 相对标准偏差(relative standard deviation, RSD)为0.1%~8.4%(n=6)。二甲双胍等3种化合物的定量限为500 μg/kg, 妥拉磺脲等5种化合物的定量限为250 μg/kg, 苯乙双胍等20种化合物的定量限为50 μg/kg。结论 本方法灵敏、高效, 可用于测定保健食品中降糖类非法添加化合物含量。

英文摘要:

      Objective To establish an analytical method for the determination of 28 adulterated anti-diabetic compounds in health foods by ultra performance liquid chromatography-triple quadrupole mass spectrometry. Methods A Merck SeQuant ZIC-HILIC column (100 mm×2.1 mm, 3.5 μm) was used with acetonitrile-0.1% formic acid aqueous solution containing 10 mmol/L ammonium formate as mobile phases by gradient elution for separating 3 compounds such as metformin. A Waters CORTECS T3 column (100 mm×2.1 mm, 2.7 μm) was used with acetonitrile-0.1%formicacid aqueous solution as mobile phases by gradient elution for separating 25 compounds such as phenformin. The compounds were detected by electrospray ion souce in positive or negative mode with multiple reaction monitoring (MRM) mode. Results The calibration curves showed good linearities for 3 compounds such as metformin in the range of 50~1000 μg/L, and for 5 compounds such as tolazamide in the range of 25~500 μg/L, and for 20 compounds such as phenformin in the range of 5~100 μg/L, respectively. The correlation coefficients (r) of the standard calibration curves for the 28 analytes were all above 0.99. The recoveries of the 28 compounds at spiked levels of limit of quantity (LOQ), double LOQ, 10 times of LOQ were in the range of 48.6%~134.4%, and the relative standard deviations (RSDs) were 0.1%~8.4% (n=6). The LOQs were 500 μg/kg for 3 compounds such as metformin, 250 μg/kg for 5 compounds such as tolazamide, 50 μg/kg for the other 20 compounds such as phenformin, respectively. Conclusion This method is selective, sensitive and accurate, which is suitable for detecting adulterated anti-diabetic compounds in health foods.