高效液相色谱-串联质谱法测定磷酸西格列汀中基因毒性杂质乙二胺的含量

Determination of Ethylenediamine as the Genotoxic Impurity in Sitagliptin Phosphate by High Performance Liquid Chromatography Tandem Mass Spectrometry

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0.50~33.5μg·L^(-1),检出限(3.3s/k)为0.55μg·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 0.50-33.5μg·L^(-1),with detection limit(3.3s/k)of 0.55μg·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%^(-1)03%.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%.