注射用聚乙二醇化重组假丝酵母尿酸氧化酶中有机溶剂残留量顶空气相色谱检测方法的建立及验证  

Establishment and verification of headspace gas chromatography method for organic solvent residues in PEGylated recombinant Candida uricase for injection

作　　者：高睿 王福鑫 黄海 GAO Rui;WANG Fuxin;HUANG Hai(Liaoning Inspection Examination and Certification Centre,Shenyang 110036,Liaoning Province,China)

机构地区：[1]辽宁省检验检测认证中心,辽宁沈阳110036

出　　处：《中国生物制品学杂志》2024年第4期493-496,504,共5页Chinese Journal of Biologicals

基　　金：辽宁省自然科学基金(2019-MS-185)。

摘　　要：目的 建立注射用聚乙二醇化重组假丝酵母尿酸氧化酶SSS111中异丙醇(isopropanol,IPA)、二氯甲烷(dichloromethane,DCM)和甲苯(toluene,TOL)残留量的顶空气相色谱检测方法,并进行验证。方法 采用DB-1301毛细管柱(30 m×0.53 mm,1.0μm)。检测温度:250℃,进样温度:200℃,载气:氮气,分流比:3,柱流速:3.0 mL/min,氢气流速:60 mL/min,空气流速:400 mL/min。柱温采用程序升温,初始柱温:50℃,维持12 min,以40℃/min的速度升温至200℃,维持5 min。顶空瓶的平衡温度:120℃,平衡时间:20 min。验证方法的线性范围、精密性、准确性,确定检测限和定量限。采用建立的方法检测3批SSS111供试品中IPA、DCM和TOL的残留量。结果 IPA、DCM、TOL分别为0.001 982~7.928、0.002 118~8.472和0.041 960~8.392μg/mL时,与峰面积呈良好的线性关系,r均> 0.99;重复6次检测对照品溶液中IPA、DCM、TOL残留量的RSD均<5%;SSS111供试品中添加对照品溶液后检测IPA、DCM、TOL回收率均在80%~120%范围内;IPA、DCM、TOL的检测限分别为1.982、2.118、41.96 ng/mL,定量限分别为6.941、6.236、126.92 ng/mL。3批供试品溶液中的IPA残留量均低于检测限,DCM残留量约为1 ppm,TOL均<1 ppm。结论 建立的顶空气相色谱检测法具有良好的精密性、准确性,且操作简便,灵敏度较高,可用于SSS111中有机溶剂残留量的检测。Objective To establish and verify a headspace gas chromatography method for the determination of residual solvents,isopropanol(IPA),dichloromethane(DCM)and toluene(TOL),in PEGylated recombinant Candida uricase for injection(SSS111).Methods A headspace gas chromatography was established using a capillary column DB-1301(30 m×0.53 mm,1.0μm),with the detector temperature of 250℃,the injector temperature of 200 C,the carrier gas of nitrogen,the split ratio of three,the column flow rate of 3.O mL/min,the hydrogen flow rate of 60 mL/min,and the air flow rate of 400 mL/min.The column temperature was programmed raising:the initial temperature was 50℃,maintaining for 12 min,and rose to 200 C at the rate of 40 C/min,maintaining for 5 min.The equilibrium temperature of the headspace bottle was 120 C,and the equilibrium time was 20 min.The linear range,precision and accuracy of the method were verified,and the limit of detection(LOD)and limit of quantitation(LOQ)were determined.IPA,DCM and TOL residues in three batches of SSS111 samples were detected by using the established method.Results IPA,DCM and TOL showed a good linear relationship with the peak area in the range of 0.001982-7.928,0.002118-8.472 and 0.041960-8.392μg/mL respectively,with each r of more than O.99.The RSDs of IPA,DCM and TOL residues in the control solution in six repeated detections were all lower than 5%.The recoveries of IPA,DCM and TOL were all in the range of 80%-120%after control solution was added to SSS111 sample.The LODs of IPA,DCM and TOL were 1.982,2.118 and 41.96 ng/mL,and the LOQs were 6.941,6.236 and 126.92 ng/mL,respectively.The residues of IPA in the three batches of samples were all lower than the LOD,the residues of DCM were about 1 ppm,and the residues of TOL were all less than 1 ppm.Conclusion The established headspace gas chromatography method has good precision and accuracy,simple operation and high sensitivity,and can be used for the determination of organic solvent residues in SSS1ll.

关 键 词：注射用聚乙二醇化重组假丝酵母尿酸氧化酶 顶空气相色谱法 有机溶剂 异丙醇 二氯甲烷 甲苯 

分 类 号：R927.11[医药卫生—药学]