乙型肝炎疫苗中聚乙二醇6000残留量HPLC-蒸发光散射检测器检测方法的建立及其验证  

Development and verification of HPLC-evaporative light scattering detector method for determination of residual polyethylene glycol 6000 content in hepatitis B vaccine

作　　者：王瑾[1] 何鹏[1] 赵冉[2] 柴龙龙 王文丽 邹烨宁 李晓东[1] 胡忠玉[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]北京科兴生物制品有限公司,北京100085 [3]淄博市食品药品检验所,山东淄博255086 [4]甘肃省药品检验研究院,甘肃兰州730070

出　　处：《中国生物制品学杂志》2017年第7期742-746,750,共6页Chinese Journal of Biologicals

基　　金：国家十二.五科学技术重大专项(2012ZX10004701)

摘　　要：目的建立测定乙型肝炎疫苗中残留聚乙二醇(PEG)6000含量的HPLC-蒸发光散射检测器(evaporative light scattering detector,ELSD)法,并进行验证。方法对ELSD条件进行优化后,按照色谱条件[色谱柱:TSK-GEL G2000-SWXL(5μm,125魡,7.8 mm×300 mm);流动相:0.025 mol/L醋酸铵溶液;流速:0.8 ml/min]上HPLC测定PEG6000标准溶液和样品中PEG6000含量,并考察该方法的线性、检出限、定量限、精密度、稳定性、回收率和适用性。结果优化后的ELSD条件为:雾化室温度40℃,漂移管温度70℃,气体流速1.20 L/min。PEG6000含量在21.6~216μg/ml范围内,该方法线性关系良好,r=0.999 5,最低检出限为2.16μg/ml,最低定量限为7.13μg/ml;PEG6000标准溶液连续检测6次,RSD为0.72%;样品溶液置不同时间检测结果的RSD<2%;不同浓度PEG6000标准溶液检测结果的回收率在97.3%~107.7%之间;使用建立的方法所有样品均未检出PEG6000,样品中含有Tween-20不干扰PEG6000的测定,分离良好。结论该方法操作简便,灵敏度高,结果准确,适用于乙型肝炎疫苗中残留PEG6000的检测。Objective To develop and verify a HPLC-evaporative-light-scattering detector （ELSD） method for determination of residual polyethylene glycol （PEG） 6000 content in hepatitis B （HB） vaccine. Methods The condition for ELSD was optimized, based on which HPLC was used for determination of PEG6000 content in standard and samples using TSK- GEL G2000SWXL column （5μm, 125 A, 7. 8 mm × 300 mm）, serving 0. 025 mol/L ammonium acetate solution as mobile phase at a flow rate of 0. 8 ml / min. The method was evaluated for linearity, limit of detection （LOD）, limit of quantitation （LOQ）, precision, stability, recovery rate and applicability. Results The optimal temperatures of atomizer chamber and drift tube for ELSD were 40 and 70 ℃ respectively, while the optimal flow rate was 1. 20 L / min. The developed method showed good linearity at a PEG6000 content range of 21. 6 - 216 μg/ml （r = 0. 999 5 ）, of which the LOD and LOQ were 2. 16 and 7. 13 μg/ml respectively. The RSD of determination results of standard PEG6000 in six consecutive tests was 0. 72%, while that of determination results at various time points was less than 2%. The recovery rates of standard PEG solutions＇at various concentrations were 97.3% - 107.7%. No PEG6000 was detected in any samples by the developed method, and the Tween-20 in samples showed no interference to the separation of PEG6000. Conclusion The method was simple, sensitive and accurate, which was suitable for the control of residual PEG6000 in HB vaccine.

关 键 词：乙型肝炎疫苗 聚乙二醇6000 HPLC 蒸发光散射检测器 

分 类 号：O657.02[理学—分析化学] R392-33[理学—化学]