HPLC-DAD法检测降糖类保健食品中非法添加的二肽基肽酶-4抑制剂和钠-葡萄糖协同转运蛋白2抑制剂

Determination of dipeptidyl peptidase-4 inhibitor and sodium glucose cotransporter 2 inhibitor illegally added in antidiabetic dietary supplements by HPLC-DAD

目的利用高效液相色谱-二极管阵列检测器(HPLC-DAD)技术,建立降糖类保健食品中可能添加的维格列汀、沙格列汀、阿格列汀、利格列汀、恩格列净、达格列净、卡格列净和西格列汀等8种化学药物的检查方法。方法采用Agilent Zorbax C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.01 mol·L^(-1)磷酸二氢铵水溶液(含0.04%辛烷磺酸钠,用磷酸调pH至3.3)为流动相,进行梯度或等度洗脱,流速1.0 mL·min^(-1),柱温25℃,检测波长为210、220 nm。结果8种化学成分在对应质量浓度范围内与峰面积呈良好的线性关系(r≥0.9999);低、中、高3个质量浓度的平均回收率均在98.8%~100.7%,RSD在0.44%~0.76%;精密度和稳定性良好,RSD均小于1.0%;检测限为0.022~0.54μg·mL^(-1);定量限为0.070~1.7μg·mL^(-1)。结论该方法简便、准确、易于推广,适用于降糖类保健食品中8种可能非法添加的新型降糖化学药物的检测。

Objective To determine the 8 chemical drugs(vildagliptin,saxagliptin,alogliptin,linagliptin,empagliflozin,dapagliflozin,canagliflozin and sitagliptin)illegally added to antidiabetic dietary supplements by high performance liquid chromatography(HPLC)method with diode array detector(DAD).Methods An Agilent Zorbax C_(18) column(250 mm×4.6 mm,5μm)was used for the separation,with gradient elution or isocratic elution of acetonitrile and aqueous solution of ammonium dihydrogen phosphate(containing 0.04%sodium octane sulfonate and adjusting pH to 3.3 with phosphoric acid)as the mobile phase at the flow rate of 1.0 mL·min^(-1).The column temperature was 25℃,and the detection wavelength was set at 210 and 220 nm.Results Totally 8 chemical constituents showed good linearity between certain mass concentration ranges and the peak area(r≥0.9999).The average spiked recoveries of the low,middle and high levels ranged 98.8%~100.7%(n=9),and the relative standard deviations(RSDs)ranged 0.44%~0.76%.The precisions and stabilities were satisfactory with both RSDs less than 1.0%.Limit of detection and limit of quantity were 0.022~0.54μg·mL^(-1)and 0.070~1.7μg·mL^(-1),respectively.Conclusion The method is simple,accurate and feasible in the detection of 8 new hypoglycemic chemical drugs illegally added in antidiabetic dietary supplements.