同位素稀释液相色谱串联质谱测定血清糖化白蛋白  

Establishment of a candidate reference method for serum glycated albumin based on liquid chromatography tandem mass spectrometry

作　　者：周慧娟 张天娇[2] 龙琪琛 曾洁[2] 张江涛[2] 周伟燕[2] 张超 张传宝[2] Zhou Huijuan;Zhang Tianjiao;Long Qichen;Zeng Jie;Zhang Jiangtao;Zhou Weiyan;Zhang Chao;Zhang Chuanbao(National Center for Clinical Laboratories,Beijing Engineering Research Center of Laboratory Medicine,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100730,China;National Center for Clinical Laboratories,Beijing Engineering Research Center of Laboratory Medicine,Beijing Hospital,National Center of Gerontology,Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]北京医院/国家老年医学中心/国家卫生健康委临床检验中心/北京市临床检验工程技术研究中心/中国医学科学院老年医学研究院/中国医学科学院北京协和医学院,北京100730 [2]北京医院/国家老年医学中心/国家卫生健康委临床检验中心/北京市临床检验工程技术研究中心/中国医学科学院老年医学研究院,北京100730

出　　处：《中华检验医学杂志》2020年第10期984-989,共6页Chinese Journal of Laboratory Medicine

摘　　要：目的建立一种基于同位素稀释液相色谱串联质谱法(ID-LC/MS/MS)检测血清糖化白蛋白(GA)的准确定量方法。方法本研究在日本临床化学学会(JSCC)推荐的ID-LC/MS方法上进行优化,采用阴离子交换色谱柱提纯白蛋白(Alb),使用重量法准确称量提纯标本、赖氨酸(Lys)和糖化赖氨酸(DOF-Lys)的标准溶液及两种的混合内标,将Alb酸水解为氨基酸,检测Lys和DOF-Lys从而定值GA。根据C62-A文件对建立的方法进行方法学评价并与常规方法进行比对研究。结果蛋白电泳鉴定标本Alb纯度结果为100%。Lys和DOF-Lys线性标准曲线R2范围分别为0.9978~0.9999、0.9994~1.0000。血清GA测定的批内、批间和总的变异系数(CV)分别为0.69%~1.97%、1.13%~2.33%、1.37%~2.54%。正确度评价中三个浓度参考物质测定结果与认定值偏差分别为-2.56%、-1.87%、-2.94%。Lys和DOF-Lys的基质效应分别为89.92%、109.98%,携带污染率分别为-1.13%、-3.27%。Lys的检测限和定量限分别为0.075、0.248 nmol/g,DOF-Lys的检测限和定量限分别为0.007、0.024 nmol/g。方法的相对扩展不确定度为1.60%~2.03%。与常规方法的拟合回归曲线R2为0.9707。结论建立了一种基于ID-LC/MS/MS法检测血清GA的方法,方法学评价准确可靠,有望作为血清GA检测的候选参考方法。Objective To establish an accurate quantitative method for the determination of serum glycated albumin(GA)based on isotope dilution liquid chromatography tandem mass spectrometry(ID-LC/MS/MS).Methods This study optimized the ID-LC/MS method recommended by the Japan Society of Clinical Chemistry(JSCC).Albumin(Alb)was purified by anion exchange chromatography.The standard solution of purified sample,lysine(Lys)and deoxyfructosyl lysine(DOF-Lys)and their mixed internal standard were weighed accurately by weight method.The Alb was hydrolyzed to amino acid,and Lys and DOF-Lys were detected to determine GA.The method was evaluated and compared with the routine method.Results The purity of Alb was 100%.The R2 ranges of Lys and DOF-Lys linear standard curves were 0.9978-0.9999 and 0.9994-1.0000.The results of three-day precision evaluation of low,medium and high concentrations of serum were 0.69%-1.97%,1.13%-2.33%and 1.37%-2.54%in intra,inter and total Coefficient of variation(CV).In the accuracy evaluation,the deviations between the results of three concentration standards and the certified value were-2.56%,-1.87%and-2.94%.The matrix effects of Lys and DOF-Lys were 89.92%and 109.98%,and the carryover rates were-1.13%and-3.27%respectively.The detection limit and quantitative limit of Lys were 0.075 nmol/g and 0.248 nmol/g,and the DOF-Lys were 0.007 nmol/g and 0.024 nmol/g.The relative expanded uncertainty was 1.60%-2.03%.The fitting regression curve R2 with the routine method was 0.9707.Conclusions A method was established for the detection of serum GA.The method evaluation was accurate and reliable.It was expected to be a candidate reference method for the detection of serum GA.

关 键 词：血清糖化白蛋白 同位素稀释液相色谱串联质谱 糖化赖氨酸 

分 类 号：R446.1[医药卫生—诊断学] O657.63[医药卫生—临床医学]