Determination of homocysteine thiolactone in human urine by capillary zone electrophoresis and single drop microextraction.

Determination of homocysteine thiolactone in human urine by capillary zone electrophoresis and single drop microextraction.
Anal Biochem. 2020 Feb 21;:113640
Authors: Purgat K, Olejarz P, Kośka I, Głowacki R, Kubalczyk P
Abstract
A simple, fast, sensitive and reproducible capillary zone electrophoresis (CZE) method with single drop microextraction (SDME) for determination of homocysteine thiolactone (HTL) in human urine has been developed and validated. The method is characterized by good precision, high accuracy, short analysis time and low consumption of reagents. The procedure consists only of few steps: urine sample centrifugation, dilution with phosphate buffer and methanol, chloroform addition onto the top of donor phase, on-line SDME in CE system, sample separation by CZE and ultraviolet detection of HTL at 240 nm. The background electrolyte was 0.1 M pH 4.75 phosphate buffer. Effective separation was achieved within 6.04 min under the separation voltage of 24 kV (∼110 μA). The LOQ and LOD for HTL were 50 and 25 nM urine, respectively. The calibration curve in urine showed linearity in the range of 50-200 nM, with R2 0.9995. The intra- and inter-day precision and recovery were 4.0-14.5% (average 8.7% and 9.3%) and 92.7-115.5% (average 103.6% and 104.8%), respectively. The procedure was successfully applied to analysis of urine samples.
PMID: 32092290 [PubMed - as supplied by publisher]

Source: Analytical Biochemistry - February 20, 2020 Category: Biochemistry Authors: Purgat K, Olejarz P, Kośka I, Głowacki R, Kubalczyk P Tags: Anal Biochem Source Type: research