当前位置： SCI文献检索 > CLINICAL CHEMISTRY期刊下所有文献 > Measurement of urinary D- and L-2-hydroxyglutarate enantiomers by stable-isotope-dilution liquid chromatography-tandem mass spectrometry after derivatization with diacetyl-L-tartaric anhydride.

Measurement of urinary D- and L-2-hydroxyglutarate enantiomers by stable-isotope-dilution liquid chromatography-tandem mass spectrometry after derivatization with diacetyl-L-tartaric anhydride.

Abstract：

BACKGROUND:The differential diagnosis of D-2-hydroxyglutaric aciduria (D-2-HGA), L-2-hydroxyglutaric aciduria (L-2-HGA), and the combined D/L-2-hydroxyglutaric aciduria (D/L-2-HGA) can be accomplished only by the measurement of the corresponding 2-hydroxyglutarate (2-HG). Available methods for the determination of D- and L-2-HG in urine are either time-consuming and expensive or have not been extensively validated. We aimed to develop a method for their rapid and sensitive measurement. METHODS:We used liquid chromatography-tandem mass spectrometry (LC-MS/MS) for the determination of D- and L-2-HG with stable-isotope-labeled internal standards. Urine samples of 20 microL were mixed with 250 microL of methanol containing the internal standards and subsequently dried under nitrogen. The analytes were derivatized by use of diacetyl-L-tartaric anhydride (DATAN) to obtain diastereomers, which were separated on an achiral C18 HPLC column and detected by MS/MS in multiple-reaction-monitoring mode. RESULTS:The use of DATAN as chiral derivatization reagent provided very well separated peaks of the formed diastereomers of D- and L-2-HG, with a total runtime of 5 min. The inter- and intraassay CVs for D- and L-2-HG ranged from 3.4% to 6.2%. Mean recoveries of D- and L-2-HG, evaluated on two concentrations, were 94%. Detection limit of the presented method was 20 pmol for a sample volume of 20 microL. Method comparison of the LC-MS/MS method with a gas chromatography-mass spectrometry method, in which D- and L-2-HG were derivatized with R-(-)-butanol, showed good agreement between the two methods. CONCLUSIONS:Urinary D- and L-2-HG can be analyzed by MS/MS after derivatization with DATAN. The presented method may be suitable for the differential diagnosis of 2-HGA.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Struys EA,Jansen EE,Verhoeven NM,Jakobs C

doi

10.1373/clinchem.2004.033399

subject

Has Abstract

pub_date

2004-08-01 00:00:00

pages

1391-5

issue

eissn

0009-9147

issn

1530-8561

pii

clinchem.2004.033399

journal_volume

pub_type

杂志文章

在线工具

Measurement of urinary D- and L-2-hydroxyglutarate enantiomers by stable-isotope-dilution liquid chromatography-tandem mass spectrometry after derivatization with diacetyl-L-tartaric anhydride.