LC-QTOF/MS法测定他达拉非中3个潜在基因毒性杂质

Determination of three potential genotoxic impurities in tadalafil by LC-QTOF/MS

目的:建立一种采用LC-QTOF/MS同时测定他达拉非原料中3个潜在基因毒性杂质:氯乙酰氯(JD-1)、(1R,3R)-1-(1,3-苯并二氧戊环-5-基)-2-(氯乙酰基)-2,3,4,9-四氢-1H-吡啶并[3,4-b]吲哚-3-羧酸甲酯(JD-2)和(1S,3R)-1-(1,3-苯并二氧戊环-5-基)-2-(氯乙酰基)-2,3,4,9-四氢-1H-吡啶并[3,4-b]吲哚-3-羧酸甲酯(JD-3)含量的方法。方法:采用苯胺为衍生化试剂,对样品进行前处理,采用YMC-PackODS-AQ色谱柱进行分离,采用乙腈和水体系为流动相,进行梯度洗脱,流速为1.0 mL·min^(-1)。质谱采用电喷雾离子源,负离子模式下选择[M-H]^(-)m/z 168.0500和[M-H]^(-)m/z 425.0855作为JD-1和JD-2、JD-3的定量离子。结果:3个潜在基因毒性杂质在37.5~112.5 ng·mL^(-1)范围内线性关系良好(r>0.9900),JD-1、JD-2和JD-3的定量限(LOQ)分别为3.8、7.3和4.1 ng·mL^(-1),3个潜在基因毒性杂质的平均回收率均在80%~120%范围内,重复性试验含量RSD均小于10%。3批原料中JD-2的含量分别为19.5、22.1、20.0μg·g^(-1),JD-3的含量分别为21.9、10.1、16.3μg·g^(-1),3批原料中均未检出JD-1。结论:该方法结果可靠,可用于他达拉非原料中3个潜在基因毒性杂质JD-1、JD-2、JD-3的含量测定。

Objective:To establish a method was established for the determination of three potential genotoxic impurities,chloroacetyl chloride(JD-1)and(1R,3R)-1-(1,3-benzodioxol-5-yl)-2-(chloroacetyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester(JD-2)and(1S,3R)-1-(1,3-benzodioxol-5-yl)-2-(chloroacetyl)-2,3,4,9-tetrahydro-1H-pyrido[3,4-b]indole-3-carboxylic acid methyl ester(JD-3)in tadalafil by LC-QTOF/MS.Methods:The samples were pretreated with aniline as a derivatization reagent.The separation was performed on a YMC-Pack ODS-AQ column withthe mobile phase consisting of water and acetonitrile by gradient elution at a flow rate of 1.0 mL·min^(-1).Electronic spray ion(ESI)in negative ionization mode was performed.The[M-H]^(-)ion(m/z 168.0500)was selected as thequantitative ion of JD-1 and the[M-H]^(-)ion(m/z 425.0855)was selected as the quantitative ion of JD-2 and JD-3.Results:The calibration curves of three potential genotoxic impurities were in good linearity(r>0.9900)within the range of 37.5-112.5 ng·mL^(-1).The lower limits of quantification(LOQ)of JD-1,JD-2 and JD-3 were 3.8,9.9 and 4.1 ng·mL^(-1),respectively.The average recoveries of the three impurities were in the range of 80%-120%and the RSDs of repeatability tests were less than 10.0%.Conclusion:The method is accurate and reliable to determine three potential genotoxic impurities JD-1,JD-2 and JD-3 in tadalafil.