通过对甲苯磺酰氯对乙二胺进行衍生化,采用高效液相色谱-串联质谱法(HPLC-MS/MS)测定磷酸西格列汀中基因毒性杂质乙二胺的含量。以Waters CORTECS^® C₁₈色谱柱为固定相,以不同体积比的5 mmol·L^-1乙酸铵-乙酸缓冲溶液(pH 5.6)和乙腈的混合溶液为流动相进行梯度洗脱。串联质谱分析中采用电喷雾离子源正离子(ESI⁺)模式,选择反应监测(SRM)模式。乙二胺的线性范围为0.50~33.5 μg·L^-1,检出限(3.3s/k)为0.55 μg·L^-1,以空白样品为基体进行加标回收试验,所得回收率为102%~103%。对平行配制的6份加标供试品溶液进行重复性和重现性试验,所得测定值的相对标准偏差均小于3.0%。

A method of high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) was proposed to determine ethylenediamine, a genotoxic impurity in sitagliptin phosphate, by derivatization of ethylenediamine with p-toluenesulfonyl chloride. Waters CORTECS^® C₁₈ column was used as stationary phase, and the mixture of 5 mmol·L^-1 ammonium acetate-acetic acid buffer solution (pH 5.6) and acetonitrile at different volume ratios was used as mobile phase for gradient elution. Electric spray ion source in positive ion mode (ESI⁺) and select reaction monitoring (SRM) were adopted in MS/MS analysis. Linear ranges of ethylenediamine was found in the range of 0.50-33.5 μg·L^-1, with detection limit (3.3s/k) of 0.55 μg·L^-1. Test for recovery was made by standard addition method on the matrix of blank sample, giving the results in the range of 102%-103%. Repeatability and reproducibility tests were carried out on the 6 spiked test solution prepared in parallel, and RSDs of the determined values were less than 3.0%.