The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine
Two cheap, simple and reproducible methods for the electrophoretic determination of homocysteine thiolactone (HTL) in human urine have been developed and validated. The first method utilizes off-line single drop microextraction (SDME), whereas the second one uses off-line SDME in combination with fi...
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doaj-a48cd1e5d99c40a0ab58c214c91808f92021-09-26T00:47:11ZengMDPI AGMolecules1420-30492021-09-01265687568710.3390/molecules26185687The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in UrineKrystian Purgat0Izabella Kośka1Paweł Kubalczyk2Department of Environmental Chemistry, Faculty of Chemistry, University of Lodz, 163 Pomorska Str., 90-236 Lodz, PolandDepartment of Environmental Chemistry, Faculty of Chemistry, University of Lodz, 163 Pomorska Str., 90-236 Lodz, PolandDepartment of Environmental Chemistry, Faculty of Chemistry, University of Lodz, 163 Pomorska Str., 90-236 Lodz, PolandTwo cheap, simple and reproducible methods for the electrophoretic determination of homocysteine thiolactone (HTL) in human urine have been developed and validated. The first method utilizes off-line single drop microextraction (SDME), whereas the second one uses off-line SDME in combination with field amplified sample injection (FASI). The off-line SDME protocol consists of the following steps: urine dilution with 0.2 mol/L, pH 8.2 phosphate buffer (1:2, <i>v/v</i>), chloroform addition, drop formation and extraction of HTL. The pre-concentration of HTL inside a separation capillary was performed by FASI. For sample separation, the 0.1 mol/L pH 4.75 phosphate buffer served as the background electrolyte, and HTL was detected at 240 nm. A standard fused-silica capillary (effective length 55.5 cm, 75 μm id) and a separation voltage of 21 kV (~99 μA) were used. Electrophoretic separation was completed within 7 min, whereas the LOD and LOQ for HTL were 0.04 and 0.1 μmol/L urine, respectively. The calibration curve in urine was linear in the range of 0.1–0.5 μmol/L, with R<sup>2</sup> = 0.9991. The relative standard deviation of the points of the calibration curve varied from 2.4% to 14.9%. The intra- and inter-day precision and recovery were 6.4–10.2% (average 6.0% and 6.7%) and 94.9–102.7% (average 99.7% and 99.5%), respectively. The analytical procedure was successfully applied to the analysis of spiked urine samples obtained from apparently healthy volunteers.https://www.mdpi.com/1420-3049/26/18/5687capillary electrophoresisfield amplified sample injectionhomocysteine thiolactoneoff-line single drop microextractionurine |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Krystian Purgat Izabella Kośka Paweł Kubalczyk |
spellingShingle |
Krystian Purgat Izabella Kośka Paweł Kubalczyk The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine Molecules capillary electrophoresis field amplified sample injection homocysteine thiolactone off-line single drop microextraction urine |
author_facet |
Krystian Purgat Izabella Kośka Paweł Kubalczyk |
author_sort |
Krystian Purgat |
title |
The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine |
title_short |
The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine |
title_full |
The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine |
title_fullStr |
The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine |
title_full_unstemmed |
The Use of Single Drop Microextraction and Field Amplified Sample Injection for CZE Determination of Homocysteine Thiolactone in Urine |
title_sort |
use of single drop microextraction and field amplified sample injection for cze determination of homocysteine thiolactone in urine |
publisher |
MDPI AG |
series |
Molecules |
issn |
1420-3049 |
publishDate |
2021-09-01 |
description |
Two cheap, simple and reproducible methods for the electrophoretic determination of homocysteine thiolactone (HTL) in human urine have been developed and validated. The first method utilizes off-line single drop microextraction (SDME), whereas the second one uses off-line SDME in combination with field amplified sample injection (FASI). The off-line SDME protocol consists of the following steps: urine dilution with 0.2 mol/L, pH 8.2 phosphate buffer (1:2, <i>v/v</i>), chloroform addition, drop formation and extraction of HTL. The pre-concentration of HTL inside a separation capillary was performed by FASI. For sample separation, the 0.1 mol/L pH 4.75 phosphate buffer served as the background electrolyte, and HTL was detected at 240 nm. A standard fused-silica capillary (effective length 55.5 cm, 75 μm id) and a separation voltage of 21 kV (~99 μA) were used. Electrophoretic separation was completed within 7 min, whereas the LOD and LOQ for HTL were 0.04 and 0.1 μmol/L urine, respectively. The calibration curve in urine was linear in the range of 0.1–0.5 μmol/L, with R<sup>2</sup> = 0.9991. The relative standard deviation of the points of the calibration curve varied from 2.4% to 14.9%. The intra- and inter-day precision and recovery were 6.4–10.2% (average 6.0% and 6.7%) and 94.9–102.7% (average 99.7% and 99.5%), respectively. The analytical procedure was successfully applied to the analysis of spiked urine samples obtained from apparently healthy volunteers. |
topic |
capillary electrophoresis field amplified sample injection homocysteine thiolactone off-line single drop microextraction urine |
url |
https://www.mdpi.com/1420-3049/26/18/5687 |
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