摘要
目的建立同时测定寡核苷酸原料药中杂质醋酸(MCAA)、二氯乙酸(DCAA)、氯离子(Cl-)含量的固相萃取离子色谱(IC)法。方法 Ionpac AS 23(4 mm×250 mm)分析柱;Ionpac AG 23(4 mm×50 mm)保护柱;流动相:0.8 mmol.L-1碳酸氢钠/4.5 mmol.L-1碳酸钠;抑制型电导检测器。结果 MCAA、DCAA和Cl-的标准曲线方程分别为:Y=5.230 7X+0.014(r=0.999 7),Y=19.762X-0.007 2(r=0.999 9)和Y=507.4X-0.746 2(r=0.999 8);MCAA在0.002～0.400μg.mL-1浓度范围内呈良好的线性关系,DCAA在0.000 624～0.156μg.mL-1浓度范围内呈良好的线性关系,Cl-在2.4×10-3～1.5 mg.mL-1浓度范围内呈良好的线性关系,其检测限分别为0.02,0.312 ng.mL-1和0.01μg.mL-1。结论该方法操作简便、快速,结果准确,可用于反义寡核苷酸原料药中杂质的控制。
Objective An solid-phase-extraction ion chromatography was developed for determining three residual impurities such as acetic acid（MCAA）,dichloroacetic acid（DCAA） and chloride ion（Cl-） in crude oligonucleotide simultaneously.Methods The final optimized conditions for testing were listed as follows by using Ionpac AS 23（4 mm×250 mm） analytical column,Ionpac AG23（4 mm×50 mm） protective column,mobile phase as 0.8 mmol·L-1 NaHCO3/4.5 mmol·L-1 Na2CO3 and suppressed conductivity detector.Results The standard curve equation of MCAA,DCAA and Cl-were Y= 5.230 7X＋0.014（r=0.999 7）,Y=19.762X-0.007 2（r=0.999 9） and Y=507.4X-0.746 2（r=0.999 8）,respectively;MCAA,DCAA and Cl-showed a good linearity in 0.002-0.400 μg·mL-1,0.000 624-0.156 μg·mL-1,0.002 4-1.5 mg·mL-1 and detection limits were 0.02,0.312 ng·mL-1 and 0.01 μg · mL-1,respectively.Conclusion The method is simple,fast,accuracy and can be used for impurities control of crude oligonucleotide.
出处
《医药导报》
CAS
北大核心
2012年第2期199-202,共4页
Herald of Medicine
基金
"十二五"国家重大新药创制科技重大专项(2011ZX09102-001-07)
"十二五"国家重大新药创制科技重大专项(2011ZX09102-009-09)
关键词
反义寡核苷酸
离子色谱法
杂质
醋酸
二氯乙酸
氯离子
Antisense oligonucleotides Ion-chromatography Impurities Acetic acid Dichloroacetic acid Chlorideion
