摘要
目的:采用二维液相色谱法检测注射用核糖核酸Ⅰ/Ⅱ/Ⅲ中苯酚的残留量,并对方法进行验证。方法:一维色谱采用ACQUITY UPLC BEH200SEC(150 mm×4.6 mm,1.7μm;填料:亚乙基桥杂化二醇包膜颗粒)色谱柱,流动相为0.1 mol·L^(-1)磷酸二氢钠溶液(p H 7.0),流速0.2mL·min^(-1),检测波长267 nm,柱温30℃;二维色谱采用Inertsil ODS SP(4.6 mm×250 mm,5μm;填料:高纯硅胶)色谱柱,以ZORBAX SB-Aq保护柱(4.6 mm×20 mm,5μm;填料:经二异丙基修饰的硅氧烷键合硅胶)作为富集柱,流动相为25%甲醇,流速1.0mL·min^(-1),检测波长267 nm,柱温30℃。检测核糖核酸(RNA)样品分别为来自健康猪肝脏的核糖核酸Ⅰ,来源于牛胰脏的核糖核酸Ⅱ,以及来源于猪脾脏的核糖核酸Ⅲ。结果:苯酚质量浓度在0.246 6~2.219 4μg·mL^(-1)范围内与峰面积呈良好线性(r=0.999 7),回收率在99.7%~100.3%;企业A的注射用核糖核酸Ⅰ和企业D的注射用核糖核酸Ⅲ样品中均未检出苯酚,企业B的注射用核糖核酸Ⅰ样品中苯酚残留量为0.002%,企业C的注射用核糖核酸Ⅱ样品中苯酚残留量为0.008%。结论:本实验建立的二维液相色谱方法能用于注射用核糖核酸Ⅰ/Ⅱ/Ⅲ中苯酚残留量的检测。
Objective：To determine the residual phenol content in ribonucleic acid Ⅰ/Ⅱ/Ⅲ for injection by two-dimensional liquid chromatography,and to validate the method.Methods：In the first dimensional chromatography,an ACQUITY UPLC BEH200 SEC（150 mm×4.6 mm,1.7 μm;column packing：ethylene bridged hybrid diol coated particles）column was used as stationary phase and 0.1 mol·L（-1） sodium dihydrogen phosphate（p H 7.0）as the mobile phase;the flow rate was 0.2mL·min（-1）;the column temperature was maintained at 30 ℃ with an ultraviolet detection of 267 nm for analysis.The target peak from the first dimensional LC was trapped in ZORBAX SB-Aq column（4.6 mm×20 mm,5 μm;column packing：silicone bonded silica gel modified by diisopropyl）and then was separated from pump of the second dimensional chromatography.An Inertsil ODS SP column（4.6 mm×250 mm,5 μm;column packing：high purity silica gel）was used in the second dimensional chromatography,with 25% methanol as the mobile phase,the flow rate was 1.0mL·min（-1）,the column temperature was maintained at 30 ℃ with an ultraviolet detection of 267 nm for analysis.Ribonucleic acid Ⅰ from pig liver,ribonucleic acid Ⅱ from bovine pancreas and ribonucleic acid Ⅲ from porcine pancreas were detected.Results：The calibration curve of phenol was linear in the concentration range of 0.246 6-2.219 4 μg·mL（-1）（r=0.999 7）.The recoveries were 99.7%（-1）00.3%.The phenol was not detectable in ribonucleic acid Ⅰ for injection produced by enterprise A and ribonucleic acid Ⅲ for injection produced by enterprise D.The phenol in ribonucleic acid Ⅰ for injection produced by enterprise B is 0.002%,and the phenol in ribonucleic acid Ⅱ for injection produced by enterprise C is 0.008%.Conclusion：The established two-dimensional liquid chromatographic method can be used for the determination of residual phenol content in ribonucleic acid Ⅰ/Ⅱ/Ⅲ for injection.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2017年第3期502-507,共6页
Chinese Journal of Pharmaceutical Analysis
关键词
猪肝脏核糖核酸Ⅰ
牛胰脏核糖核酸Ⅱ
猪脾脏核糖核酸Ⅲ
苯酚残留检测
二维液相色谱
ribonucleic acid Ⅰ for injection from pig liver
ribonucleic acid Ⅱ for injection from bovine pancreas
ribonucleic acid Ⅲ for injection from porcine pancreas
detection of phenol residues
two-dimensional liquid chromatography
