分析测试学报2023，Vol.42Issue(12)：1639-1644,1651,7.DOI:10.19969/j.fxcsxb.23071501
凝胶过滤层析结合液相色谱-串联质谱法测定血清中游离甲状腺激素
Determination of Free Thyroid Hormones in Serum by Gel Filtration Chromatography Combined with Liquid Chromatography-Tandem Mass Spectrometry
摘要
Abstract
A method was developed for high throughput and high sensitive detection of free triiodothy-ronine(FT3)and free thyroxine(FT4)in serum using gel filtration chromatography(GFC)and liquid chromatography-tandem mass spectrometry(LC-MS/MS).The hydroxypropyl cellulose gel Sephadex LH-20 column was used to filter and separate 150 μL serum samples.First of all,the thyroid hor-mones bound to binding proteins were removed by washing with tri(hydroxymethyl)aminomethane hy-drochloride(Tris-HCl)buffer solution(0.1 mol/L,pH 7.4).Following that,the free thyroid hor-mones were eluted using methanol.The established LC-MS/MS analysis method was employed for de-tection and quantification.The results of the methodological validation showed that the correlation co-efficient(r2)was not less than 0.999 5.The precision ranges from 1.8%to 10%,accuracy ranges from 85.4%to 110%,matrix effect ranges from 85.8%to 114%,and recoveries from 85.8%to 107%.According to the CLSI EP09-A3 method comparison guidelines,this method was further ap-plied to the detection of 95 serum samples and compared with the equilibrium dialysis(ED),to evalu-ate the accuracy and applicability of the established GFC/LC-MS/MS method.The statistical analysis was performed using IBM SPSS Statistics 26 and MedCalc statistical software(v.20.0.0)for paired t-test analysis,Passing-Bablok regression analysis,and Bland-Altman agreement analysis.The sta-tistical analysis results indicated that there was no significant difference(P>0.05)between the detec-tion results of FT3 and FT4 in the serum measured by the developed GFC/LC-MS/MS method and the ED/LC-MS/MS method.The developed method can provide a reference method for rapid,efficient,and accurate detection of free thyroid hormones in clinical laboratories,as well as offering a new ap-proach for the detection of other free hormones such as cortisol and testosterone.关键词
液相色谱-串联质谱法/游离甲状腺激素/凝胶过滤层析/血清Key words
liquid chromatography-tandem mass spectrometry/free thyroid hormones/gel filtra-tion chromatography/serum分类
化学化工引用本文复制引用
邹海飞,李敏,邓顺艳,夏兵,周燕..凝胶过滤层析结合液相色谱-串联质谱法测定血清中游离甲状腺激素[J].分析测试学报,2023,42(12):1639-1644,1651,7.基金项目
中国科学院仪器功能开发与技术创新项目(E1C3030001) （E1C3030001）
