Simultaneous determination of total homocysteine and cysteine in human plasma by capillary zone electrophoresis with pH-mediated sample stacking

Kubalczyk, Paweł; Bald, Edward; Furmaniak, Paulina; Głowacki, Rafał

doi:10.1039/c4ay00287c

Cited by 50 publications

(36 citation statements)

References 49 publications

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“…The analytical criteria (precision, accuracy) are comparable with those obtained by analogous methods. [12][13][14]21,22 We conclude that this method may find application in the routine analysis of blood plasma samples in experimental and clinical laboratories. Further improvement of the deproteinization approaches based on ultracentrifugation or solid-phase extraction may facilitate sample preparation and speed up the process.…”

Section: Discussionmentioning

confidence: 99%

“…The sensitivity and productivity of the developed approach (0.3 -0.8 μM) was comparable to that of other CE-UV methods. 13,14 The limits of this approach include the inability to determine cysteinylglycine because the mobility of this analyte is not sufficient for this method. Aminothiols that lack an acidic group are also not suitable for analysis using this methodology.…”

Section: Discussionmentioning

confidence: 99%

“…A few rather sensitive HPLC approaches (LOD < 1 μM) were also proposed in which the application of reagents, such as 2-chloro-1-methylquinoline tetrafluoroborate (CMQT), 8 1,1′-thiocarbonyldiimidazole (TCDI), 9 4,4′-dithiodipyridine 10 and 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) 11 made it possible to use conventional UV-detectors. Later approaches based on CE-UV with the use of CMQT 12,13 and TCDI 14 were proposed. Although these approaches had a lower sensitivity, they had a number of advantages compared to HPLC such as high performance, selectivity (specificity) and affordability.…”

Section: Introductionmentioning

confidence: 99%

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Determination of Blood Plasma Aminothiols Using Derivatization-enhanced Capillary Transient Isotachophoresis

et al. 2018

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Experimental Reagents and chemicalsAcetonitrile (ACN; 99.9%; Lab-Scan, Gliwice, Poland); CHCl3 (>99.8% for LC; Merck, Darmstadt, Germany); L-Cys 97%, dimethyl sulfoxide (DMSO, >99.8%), dithiothreitol (DTT, >99%), DTNB, formic acid 98%, D,L-Hcy >95%, GSH reduced >98%, N-(2-mercaptopropionyl)glycine (MPG), NaCl 99.5%, A sensitive capillary electrophoresis method was developed for the determination of aminothiol (cysteine, homocysteine, and glutathione) total levels in human blood plasma. Analytes were derivatized with Ellman's reagent (5,5′-dithiobis(2-nitrobenzoic acid)) after reduction with dithiothreitol. Liquid-liquid extraction was applied to purify the samples and concentrate the analytes. Total analysis time was 7.5 min using a silica capillary (50 μm i.d.; effective separation length 23.5 cm). Electrophoretic separation was performed using 50 mM citric acid with 20 mM triethanolamine (pH 3) containing 2% Ficoll 400. Detection limit was 0.8 μM for glutathione and 0.3 μM for both cysteine and homocysteine. Accuracy was 94 -107%, repeatability and reproducibility were ca. 2.7 -3.5 and 2.5 -6.5%, respectively.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Determination of Blood Plasma Aminothiols Using Derivatization-enhanced Capillary Transient Isotachophoresis

et al. 2018

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“…Thiosulfate unluckily does not possess structural properties enabling efficient absorption of radiation in the UV‐Vis region, thus we resort to derivatization reaction. In this procedure we used CMQT which is well known as the suitable reagent for thiols derivatization . It was earlier proved that CMQT could also effectively react with thiosulfates .…”

Section: Resultsmentioning

confidence: 99%

Fast and simple MEKC sweeping method for determination of thiosulfate in urine

2015

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A new method for determination of thiosulfate in human urine has been developed and validated. Analytical procedure is very simple and consists of only few steps: derivatization of thiosulfate with 2-chloro-1-methylquinolinium tetrafluoroborate, centrifugation of a mixture, separation of so-formed derivative by micellar electrokinetic chromatography with sweeping and UV detection at 375 nm. A fused-silica capillary with an inlet to detector length of 51.5 cm and a total length of 60 cm (75 μm id) was served as a separation column. The separation voltage of 20.5 kV (∼160 mA) and buffer solution consisting of 0.055 mol/L sodium phosphate (pH 8), 25% acetonitrile, and 0.035 mol/L sodium dodecyl sulfate were found to be the most suitable conditions for the effective separation. The limit of quantification for thiosulfate was 4 μmol/L urine. The method was validated and calibrated for thiosulfate in the range of 4-64 μmol/L (R(2) = 0.9997). The relative standard deviation of the points of the calibration curve varied from 1.2 to 4.8% RSD.

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“…It has been earlier shown that quinolinium as well as lepidinium salts are selective toward thiols affording appropriate UV absorbing derivatives [22][23][24][25][26]. Since CMLT possesses additional methyl group in quinolinium moiety, consequently, 2-S-lepidinium derivatives are more lipophilic than 2-S-quinolinium ones and therefore undergo stronger retention on stacked micelles.…”

Section: In-capillary Derivatizationmentioning

confidence: 98%

Novel MEKC method for determination of sodium 2-mercaptoethanesulfonate in human plasma with in-capillary derivatization and UV detection

Furmaniak

Kubalczyk

Stachniuk

et al. 2016

Journal of Chromatography B

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Simultaneous determination of total homocysteine and cysteine in human plasma by capillary zone electrophoresis with pH-mediated sample stacking

Cited by 50 publications

References 49 publications

Determination of Blood Plasma Aminothiols Using Derivatization-enhanced Capillary Transient Isotachophoresis

Determination of Blood Plasma Aminothiols Using Derivatization-enhanced Capillary Transient Isotachophoresis

Fast and simple MEKC sweeping method for determination of thiosulfate in urine

Novel MEKC method for determination of sodium 2-mercaptoethanesulfonate in human plasma with in-capillary derivatization and UV detection

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