柱前衍生HPLC法检测硼替佐米中间体中异戊胺盐酸盐  

Determination of the impurity of isopentylamine hydrochloride in the intermediate of bortezomib by HPLC with pre-column derivatization

作　　者：王小文 刘佳 郑秋莹 李朴超 唐海 周微 刘全礼[1] WANG Xiaowen;LIU Jia;ZHENG Qiuying;LI Puchao;TANG Hai;ZHOU Wei;LIU Quanli(College of Pharmacy,Baotou Medical College,Baotou 014060,China;Tasly Research Institute,Jiangsu TaslyDiyi Pharmaceutical Co.,Ltd,Tianjin 300410,China)

机构地区：[1]包头医学院药学院,包头014060 [2]江苏天士力帝益药业有限公司天士力研究院,天津300410

出　　处：《分析试验室》2022年第9期1089-1095,共7页Chinese Journal of Analysis Laboratory

摘　　要：建立了以二氯甲烷为反应溶剂、三乙胺为缚酸剂、苯甲酰氯为衍生试剂的柱前衍生高效液相色谱法,同时测定硼替佐米无紫外吸收的中间体(R)-1-氨基-3-甲基丁基硼酸频那醇酯盐酸盐及异戊胺盐酸盐含量的分析方法。异戊胺盐酸盐在冰浴下与衍生化试剂反应进行柱前衍生,并利用高效液相色谱-质谱对衍生产物进行定性分析。采用AgelaVenusil MP C18(2)(250 mm×4.6 mm,5μm)色谱柱,甲醇-水(70∶30,V/V)为流动相,流速为1.0 mL/min,检测波长为236 nm,柱温为35℃。异戊胺盐酸盐在质量浓度0.075~30μg/mL范围内线性关系良好(r=0.9999),定量限和检出限分别为0.075和0.030μg/mL,信噪比分别为44.6和7.8;回收率在97.7%~104.0%之间,相对标准偏差(RSD)为2.9%。该方法可用于杂质异戊胺盐酸盐的含量测定,对于其他无紫外吸收胺盐类化合物的测定有重要的参考价值。A method was developed for the simultaneous determination of(R)-pinacol-1-amino-3-methylbutylborate hydrochloride and isopentylamine hydrochloride by pre-column derivatization high performance liquid chromatography(HPLC)with dichloromethane as solvent,triethylamine as acid binding agent and benzoyl chloride as derivative reagent.(R)-pinacol-1-amino-3-methylbutylborate hydrochloride and isopentylamine hydrochloride reacted with derivatization reagent in an ice bath for 10 min.The derivatives were qualitatively determined by HPLC-mass spectrometry(HPLC-MS).HPLC was performed on an AgelaVenusil MP C18(2)column using methanol-water(70∶30,V/V)as mobile phase.The flow rate was 1.0 mL/min,the detection wavelength was 236 nm and the column temperature was 35℃.The UV absorption produced after derivatization,which could be detected by HPLC with good specificity.Isopentylamine hydrochloride derivative showed an excellent linearity in the range of 0.075-30μg/mL,with the limit of quantification and the limit of detection(LOD)of 0.075 and 0.030μg/mL,respectively.And the S/N of the LOQ and the LOD were 44.6 and 7.8,respectively.The average recoveries were 97.7%-104.0%with the RSD of 2.9%.The method is suitable for the determination of isoamylamine hydrochloride and has significant reference value for the determination of other amine salt without UV absorption.

关 键 词：高效液相色谱法 柱前衍生 胺盐 硼替佐米 异戊胺盐酸盐 

分 类 号：R917[医药卫生—药物分析学]