孕酮冰冻人血清国家标准品的研制  

Development of the national standard material of progesterone in frozen human serum

作　　者：于婷[1] 唐利萍 黄杰[1] 孙可其 YU Ting;TANG Li-ping;HUANG Jie;SUN Ke-qi(National Institutes for Food and Drug Control,Beijing 100050,China;Reference System Center of Maccura Biotechnology Co.,Ltd.,Chengdu 611731,China)

机构地区：[1]中国食品药品检定研究院,北京100050 [2]迈克生物股份有限公司参考系统中心,成都611731

出　　处：《药物分析杂志》2024年第3期547-552,共6页Chinese Journal of Pharmaceutical Analysis

基　　金：国家重点研发计划(2019YFF0216501)。

摘　　要：目的:研制孕酮冰冻人血清国家标准品,用于血清孕酮检测试剂盒的正确度评价及标准化研究。方法:分别收集来自正常人及孕妇的血清,且外观澄明、无明显溶血、黄疸和脂血,传染病四项检测均为阴性后,按比例分别添加凝血酶1 kU·L^(-1),无水氯化钙2.2196 g·L^(-1)和无水碳酸钠2.65 g·L^(-1),充分混匀后,离心(4000 r·min^(-1),4℃,25 min);再经2次多重过滤除菌后分装,制备成候选品Ⅰ和Ⅱ,-70℃条件下保存。分别采用单因素方差法和线性回归法评估候选品的均匀性和稳定性。联合5家实验室采用参考方法(同位素稀释液相色谱串联质谱法)定值,并计算不确定度。同时对候选品互通性进行评估。结果:经统计分析,候选品Ⅰ、Ⅱ的均匀性检验F值分别为0.5709和1.2009,均<F0.05;候选品Ⅰ、Ⅱ在(20~25)℃至少可稳定8 h;在(2~8)℃至少分别可稳定33 d和7 d;在-20℃至少可稳定33 d。候选品Ⅰ、Ⅱ反复冻融6次后稳定。候选品Ⅰ、Ⅱ定值结果为:Ⅰ:(4.44±0.24)ng·mL^(-1)(k=2);Ⅱ:(16.79±0.82)ng·mL^(-1)(k=2)。候选品Ⅰ、Ⅱ测定结果均在新鲜血清样本测定值所建立的拟合回归直线95%置信区间范围内。结论:孕酮候选品均匀性、稳定性、互通性良好,定值准确可靠,可作为国家标准品应用。该国家标准品的建立,对促进孕酮检测结果的一致化和标准化具有重要意义。Objective:To develop the national standard materials of progesterone in frozen human serum and to evaluate the accuracy and standardization of serum progesterone detection kits.Methods:Low and high concentrations of serum were from normal people and pregnant women individuals respectively,with a clear appearance,no obvious hemolysis,jaundice and lipid blood.After all tests for four infectious diseases were negative,thrombin,anhydrous calcium chloride and anhydrous sodium carbonate were added according to the proportion of1 kU·L^(-1),2.2196 g·L^(-1)and 2.65 g·L^(-1)respectively,and centrifuged(4000 r·min^(-1),4℃,25 min)after fully mixing.After twice of multiple filtration and sterilization,serum pools were packed in ampoules to prepare candidatesⅠandⅡand stored at-70℃.Single factor variance method and linear regression method were used to evaluate the homogeneity and stability of the candidate.Five labs working together used reference method(isotope dilution liquid chromatography tandem mass spectrometry)to assign value for candidate.The uncertainty was calculated.At the same time,the commutability was evaluated.Results:Through statistical analysis,the F values of homogeneity for candidateⅠandⅡwere 0.5709 and 1.2009,respectively,which were all less than F_(0.05).CandidateⅠandⅡcould be stable at least 8 h at 20-25℃.At 2-8℃,candidateⅠandⅡcould be stable at least 33 d and 7 d respectively.CandidateⅠandⅡcound be stable at least 33 d at-20℃.CandidateⅠandⅡremained stable after repeated freeze-thaw cycles for 6 times.The assigned values were:candidateⅠ:(4.44±0.24)ng·mL^(-1)(k=2),Ⅱ:(16.79±0.82)ng·mL^(-1)(k=2).The determination results of candidateⅠandⅡwere all within the 95%confidence interval of the fitting regression line established by the determination values of fresh serum samples.Conclusion:Progesterone candidate had good homogeneity,stability,interoperability.It had accurate and reliable determination value,which could be used as national standard.The establishment

关 键 词：孕酮 国家标准品 准确度评价 标准化 互通性 

分 类 号：R917[医药卫生—药物分析学]