摘要
目的建立血浆源性人凝血因子Ⅷ(plasma-derived FⅧ,pdFⅧ)制品中钙含量的原子吸收分光光度检测方法,并评价方法的不确定度,以期为血液制品中钙含量的检测提供可靠方法。方法pdFⅧ中添加电离抑制剂氯化镧后,采用原子吸收分光光度法检测钙含量,并优化方法中的燃气流量(1.2、1.4、1.6、1.8、2.0 L/min)、燃烧器高度(9、11、12、13 mm)、灯电流(100%、90%、85%、80%)、狭缝宽度(0.2、0.5、1.0 nm),验证方法的专属性、线性范围、检测限及定量限、准确性、精密性、稳定性。采用建立方法检测6批pdFⅧ供试品中钙含量。评价方法的不确定度,并以批号为202404026的pdFⅧ供试品为例,测定钙含量,确定各不确定度来源对检测结果的影响。结果确定最佳燃气流量为1.8 L/min,燃烧气高度为12 mm,灯电流为85%,狭缝宽度为0.5 nm。氯化镧不干扰pdFⅧ中钙含量的测定;钙工作标准液在1.0~5.0μg/mL浓度范围内,与A422.7呈良好的线性关系,回归方程为:y=0.0166 x-0.0008,r=0.9987;检测限为0.1272μg/mL,定量限为0.3855μg/mL;低、中、高浓度加标样品的回收率均在85%110%范围内;精密性及稳定性验证RSD均<2%。6批pdFⅧ供试品的钙含量均≤50μg/mL。不确定度主要来源于标准溶液的制备,批号为202404026的pdFⅧ供试品钙含量为(42.11±1.7361)μg/mL,符合规定(≤50μg/mL),表明各不确定度来源对钙浓度检测结果未产生影响。结论建立的原子吸收分光光度法具有良好的准确性、精密性及稳定性,且通过不确定度评价确保了检测结果的可靠性,可用于pdFⅧ中钙含量的准确测定。
Objective To establish an atomic absorption spectrophotometry for the determination of calcium content in human plasma-derived FVII(pdFVIl)products,and to evaluate the uncertainty of the method,so as to provide a reliable method for the determination of calcium content in blood products.Methods After adding lanthanum chloride solution as ionization inhibitor to pdFVIl,the calcium content was detected by atomic absorption spectrophotometry.The gas flow rate(1.2,1.4,1.6,1.8,2.0 L/min),burner height(9,11,12,13 mm),lamp current(100%,90%,85%,80%),and slit width(0.2,0.5,1.0 nm)were optimized,and the method was verified for specificity,linearity range,limit of detection(LOD),limit of quantitation(LOQ),accuracy,precision and stability.The calcium content in six batches of pdF VIl samples was determined by the established method.The uncertainty of the method was evaluated,and taking pdFVIl sample of batch number 202404026 as an example,the calcium content was determined,and the effects of various sources of uncertainty on the detection results were determined.Results The optimum gas flow rate was 1.8 L/min,the combustion gas height was 12 mm,the lamp current was 85%,and the slit width was O.5 nm.Lanthanum chloride did not interfere with the determination of calcium content in pdF VII.In the concentration range of 1.0-5.0μg/mL,the calcium working standard solution had a good linear relationship with A42.7.with the regression equation of:y=0.0166 x-0.0008,r=0.9987.The LOD was 0.1272μg/mL,and the LOQ was 0.3855μg/mL.The recovery rates of low,medium and high concentration spiked samples were all in the range of 85%-110%.The RSDs of precision and stability verification were both less than 2%.The calcium content of six batches of pdF VIl samples was not more than 50μg/mL.The uncertainty was mainly derived from the preparation of standard solution.The calcium content of pdF Ⅷ sample with batch number 202404026 was(42.11±1.7361)μg/mL,which met the requirement(≤50μg/mL),indicating that the sources of uncertainty showed no influence on the determination results of calcium concentration.Conclusion The established atomic absorption spectrophotometry has good accuracy,precision and stability,and the reliability of detection results is ensured by uncertainty evaluation,which can be used for the accurate determination of calcium content in pdFⅧ.
作者
张婷
周长明
沈琮
邵天舒
丁锐
ZHANG Ting;ZHOU Changming;SHEN Cong;SHAO Tianshu;DING Rui(Beijing Institute for Drug Control(Beijing Center for Vaccine Control),NMPA Key Laboratory for Research and Evaluation of Generic Drug,Beijing Key Laboratory of Analysis and Evaluation on Chinese Medicine,Beijing 102206,China)
出处
《中国生物制品学杂志》
2025年第5期581-587,共7页
Chinese Journal of Biologicals
基金
国家药品标准提高项目(BZ2024162)。
