干旱区生态环境知识资源中心(Arid): 人凝血因子Ⅷ制品中聚山梨酯80残留量高效液相色谱-电喷雾检测器检测方法的建立及验证

Arid

	人凝血因子Ⅷ制品中聚山梨酯80残留量高效液相色谱-电喷雾检测器检测方法的建立及验证
其他题名	Development and verification of high performance liquid chromatography coupled with charged aerosol detection for residual polysorbate 80 content in human coagulation factor Ⅷ products
	邵天舒; 丁锐; 周长明; 郭雷
来源期刊	中国生物制品学杂志
ISSN	1004-5503
出版年	2021
卷号	34 期号:9 页码:1105-1110
中文摘要	目的建立人凝血因子Ⅷ制品中聚山梨酯80残留量的高效液相色谱(HPLC)-电喷雾检测器(charged aerosol detection,CAD)检测方法,并进行验证及初步应用。方法采用反相-离子交换混合色谱柱Oasis MAX(2.1 mm *20 mm, 30 mum),流动相A为含2%(V/V)甲酸的异丙醇∶水(20 ∶ 80)溶液,流动相B为含2%(V/V)甲酸的异丙醇溶液,流速为1.0 mL/min,柱温为30 ℃,进样量为30 muL,梯度洗脱,检测人凝血因子Ⅷ中聚山梨酯80残留量。验证方法的系统适用性、线性范围、检测限、定量限、重复性、稳定性及准确性,并与HPLC-蒸发光散射检测器(evaporative light scattering detector,ELSD)检测法进行比较。应用建立的HPLC-CAD方法检测10批人凝血因子Ⅷ样品中聚山梨酯80残留量。结果聚山梨酯80与其他组分色谱峰分离度良好,无其他色谱峰干扰。经HPLC-CAD检测,聚山梨酯80标准品在12.8 ~ 256.3 mug/mL浓度范围内线性关系良好,回归方程y = 121.42 x + 1.321 7,r = 0.998 0;定量限约为3.20 mug/mL,检测限约为1.07 mug/mL;供试品溶液重复检测6次聚山梨酯80含量的RSD为0.86%;供试品溶液室温放置0、2、4、6、8及10 h聚山梨酯80含量的RSD为0.65%;浓度为32.8、41.0及49.2 mug/mL的加标样品回收率为96.85% ~ 101.49%;该方法的定量限、检测限、重复性、稳定性、准确性指标均优于HPLC-ELSD。10批样品中聚山梨酯80残留量在56.09 ~ 66.75 mug/mL,均符合《中国药典》三部(2015版)要求(≤ 100 mug/mL)。结论成功建立了检测人凝血因子Ⅷ制品中聚山梨酯80残留量的HPLC-CAD法,该方法重复性、稳定性及准确性良好,可用于检测人凝血因子Ⅷ制品中聚山梨酯80残留量。
英文摘要	Objective To develop,verify and perliminarily apply a method for determination of residual polysorbate 80 content in human coagulation factor Ⅷ(FⅧ)products by high performance liquid chromatography(HPLC)coupled with charged aerosol detector(CAD). Methods Oasis MAX column(2.1 mm *20 mm,30 mum)was adopted at a column temperature of 30 ℃. The mobile phase A was composed of isopropanol-water(20 ∶ 80)containing 2%(V/V)formic acid,while the mobile phase B was isopropanol containing 2%(V/V)formic acid,with gradient elution at a flow rate of 1.0 mL/min. The sample load was 30 muL. The developed HPLC-CAD method was verified for systemic suitability,linear range,detection limit,quantitative limit,reproducibility,stability and accuracy,by which the determination result was compared with that by HPLC-ELSD(evaporative light scattering detector)method. Results The residual polysorbate 80 showed a good separation with other components of FⅧ products,without interference of any other peaks. The linear range of polysorbate 80 was 12.8 ~ 256.3 mug/mL,with a regression equation of y = 121.42 x + 1.321 7 and a correlation coefficients(r)of 0.998 0. The quantitative limit of the developed method was about 3.20 mug/mL,while the detection limit was about 1.07 mug/mL. The RSD of determination results of polysorbate 80 contents in six repeat test was 0.86%, while that the spike recoveries in samples after storage at room temperature for 0,2,4,6,8 and 10 h was 0.65%,of polysorbate 80 in samples at concentrations of 32.8,41.0 and 49.2 mug/mL were 96.85% ~ 101.49%. The quantitative and detection limits of the developed method were lower that,while the reproducibility,stability and accuracy were superior to,those of HPLC-ELSD. The residual polysorbate 80 contents in ten batches of samples were 56.09 ~ 66.75 mug/mL, which met the requirements in Chinese Pharmacopeia(VolumeⅢ2015 deition,not more than 100 mug/mL). Conclusion The HPLC-CAD method for residual polysorbate 80 content in FⅧ products was successfully developed,which was sensitive,reproducible,stable and accurate.
中文关键词	人凝血因子Ⅷ ; 聚山梨酯80 ; 高效液相色谱 ; 电喷雾检测器
英文关键词	Human coagulation factor Ⅷ products Polysorbate 80 High performance liquid chromatography(HPLC) Charged aerosol detection(CAD)
类型	Article
语种	中文
收录类别	CSCD
WOS类目	Pharmacology & Pharmacy
CSCD记录号	CSCD:7052567
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/378126
作者单位	邵天舒, 北京市药品检验所, 国家药品监督管理局仿制药研究与评价重点实验室;;中药成分分析与生物评价北京市重点实验室, 北京 102206, 中国.; 丁锐, 北京市药品检验所, 国家药品监督管理局仿制药研究与评价重点实验室;;中药成分分析与生物评价北京市重点实验室, 北京 102206, 中国.; 周长明, 北京市药品检验所, 国家药品监督管理局仿制药研究与评价重点实验室;;中药成分分析与生物评价北京市重点实验室, 北京 102206, 中国.; 郭雷, 北京市药品检验所, 国家药品监督管理局仿制药研究与评价重点实验室;;中药成分分析与生物评价北京市重点实验室, 北京 102206, 中国.
推荐引用方式 GB/T 7714	邵天舒,丁锐,周长明,等. 人凝血因子Ⅷ制品中聚山梨酯80残留量高效液相色谱-电喷雾检测器检测方法的建立及验证[J],2021,34(9):1105-1110.
APA	邵天舒,丁锐,周长明,&郭雷.(2021).人凝血因子Ⅷ制品中聚山梨酯80残留量高效液相色谱-电喷雾检测器检测方法的建立及验证.中国生物制品学杂志,34(9),1105-1110.
MLA	邵天舒,et al."人凝血因子Ⅷ制品中聚山梨酯80残留量高效液相色谱-电喷雾检测器检测方法的建立及验证".中国生物制品学杂志 34.9(2021):1105-1110.