超高效液相色谱法同时测定降糖类保健食品中非法添加的13种化学药物

Simultaneous Determination of 13 Illegally Added Chemical Drugs in Hypoglycemic Health Foods by Ultra Performance Liquid Chromatography

建立超高效液相色谱法,同时测定降糖类保健食品中可能非法添加的13种化学药物（盐酸二甲双胍、盐酸丁二胍、盐酸苯乙双胍、甲苯磺丁脲、格列吡嗪、格列齐特、格列波脲、盐酸罗格列酮、格列本脲、盐酸吡格列酮、格列美脲、瑞格列奈、格列喹酮）。样品采用甲醇提取,经ZORBAX SB-C18色谱柱分离,乙腈和0.02 mol/L乙酸铵为流动相梯度洗脱,流速0.2 mL/min。采用二极管阵列检测器,采集波长235 nm处的色谱图进行初筛和定量。并通过与13种降糖药物标准品的光谱比对进行确证。结果表明,13种降糖药物在30 min内可有效分离,在0.3-90μg/mL的线性范围内相关系数均大于0.999,平均回收率为83.2%-108.9%,相对标准偏差均不大于5.5%,检出限为0.2-0.6 ng。该方法前处理简单、分析时间相对较短、灵敏度高且重复性好,实现一个系统对13种降糖药物的同时检测,可作为降糖类保健食品检测非法添加化学成分的参考方法。

An ultra performance liquid chromatography（UPLC） method was developed for the simultaneous determination of 13 illegally added chemical drugs（metformin hydrochloride,buformin hydrochloride, phenformin hydrochloride, tolbutamide, glipizide, gliclazide, glibornuride, rosiglitazone hydrochloride, glibenclamide, pioglitazone hydrochloride, glimepiride, repaglinide and gliquidone） in hypoglycemic health foods. Samples were extracted by using methanol. ZORBAX SB-C18 column was applied with a mobile phase consisting of acetonitrile and 0.02 mol/L ammonium acetate by gradient elution at a flow rate of 0.2 mL/min. The illegally added chemical drugs were detected by a photodiode array detector and a chromatogram with detection wavelength of 235 nm was used for screening and quantification. The results were then confirmed by comparison with the spectra of 13 reference substances. It was shown that the 13 anti-diabetic agents could be effectively separated within 30 min. The correlation coefficients were all above 0.999 over the concentration range of 0.3–90 μg/mL. The average recoveries ranged from 83.2% to 108.9% with relative stand deviations（RSDs） all below 5.5%. The limits of detection（LODs） were all in the range of 0.2 to 0.6 ng. This method has the advantages of simple pre-treatment, fast detection, satisfactory recovery rate and good reproducibility, and it is suitable for the simultaneous determination of the 13 anti-diabetic agents and can be used as a reference in the detection of the illegally added chemical drugs in hypoglycemic health foods.