A single-step method for simultaneous quantification of thiamine and its phosphate esters in whole blood sample by ultra-performance liquid chromatography-mass spectrometry

被引：12

作者：

Cheng, Xiaoqin ^{[1
]}

Ma, Dong ^{[1
,2
]}

Fei, Guoqiang ^{[1
]}

Ma, Zhengyao ^{[3
]}

Xiao, Feng ^{[4
]}

Yu, Qiujian ^{[1
]}

Pan, Xiaoli ^{[1
]}

Zhou, Feng ^{[5
]}

Zhao, Lei ^{[6
]}

Zhong, Chunjiu ^{[1
]}

机构：

[1] Fudan Univ, Collaborat Innovat Ctr Brain Sci, Inst Brain Sci, State Lab Med Neurobiol,Dept Neurol,Zhongshan Hos, Shanghai, Peoples R China

[2] Acad Forens Sci, Shanghai 200063, Peoples R China

[3] Fudan univ, Zhongshan Hosp, Dept Clin Lab, Shanghai, Peoples R China

[4] Shanghai Inst Pharmaceut Ind, Shanghai, Peoples R China

[5] Fudan Univ, Inst Biomed Sci, Shanghai, Peoples R China

[6] Shanghai Jiao Tong Univ, Shanghai Peoples Hosp 9, Sch Med, Shanghai, Peoples R China

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2018年 / 1095卷

基金：

中国国家自然科学基金;

关键词：

Thiamine diphosphate; Thiamine monophosphate; Thiamine; Ultra-performance liquid chromatography-mass spectrometry; Human whole blood; DEPENDENT PROCESSES; GLUCOSE-METABOLISM; HUMAN ERYTHROCYTES; HUMAN PLASMA; DIPHOSPHATE; DEFICIENCY; HPLC; OUTBREAK;

D O I：

10.1016/j.jchromb.2018.07.030

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Thiamine and its phosphate esters play vital physiological roles and thiamine deficiency causes deleterious effects on human body. It is important to quantify accurately the thiamine metabolites in body fluids. However, due to the lack of appropriate internal standards, poor inter-laboratory standardization and time-consuming pretreatment procedure, the existing methods are limited in clinical applications. Hence, we developed a singlestep HPLC-MS/MS method for accurate and precise measurement of thiamine and its phosphate esters in human whole blood. Whole blood samples were deproteinized and the supernatants were collected. The levels of thiamine diphosphate (TDP), thiamine monophosphate (TMP), and thiamine were determined by HPLC-MS/MS method after adding isotopic internal standards. The method was linear from 15.625-3.125-1.563 nmol/L to 1000-200-100 nmol/L for TDP-TMP-thiamine. The lower limit of quantification was 15.625-3.125-1.563 nmol/L. The intra-day and inter-day precisions and accuracy for all QCs samples were <= 15.9% and <= 11.1%, respectively. The matrix effect was not significant. Recoveries were 103.7% for TDP, 102.7% for TMP, and 105.3% for thiamine. All QCs were stable for three freeze-thaw cycles, or at room temperature for 3 h, or at - 80 degrees C for 15 days. We compared this new method with an established HPLC method based on derivatization of thiamine metabolites. It is found that this method correlated well with HPLC method for TDP determination (R-2 = 0.93). However, the correlation was not ideal for TMP (R-2 = 0.40) or thiamine (R-2 = 0.72) determination. Subject's diet was shown to have no significant effect on the concentrations of thiamine metabolites in their blood samples. To conclude, we developed a single-step, non-derivatization HPLC-MS/MS method that can detect thiamine and its phosphate esters in human whole blood accurately and quickly.

引用

页码：103 / 111

页数：9

共 37 条

[1] Outbreak of beriberi in a prison in Cote d'Ivoire [J].