Ultraviolet derivatization of low-molecular-mass thiols for high performance liquid chromatography and capillary electrophoresis analysis

Kuśmierek, Krzysztof; Chwatko, Grażyna; Głowacki, Rafał; Kubalczyk, Paweł; Bald, Edward

doi:10.1016/j.jchromb.2010.10.035

Cited by 111 publications

(98 citation statements)

References 148 publications

(189 reference statements)

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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: 81%

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: 81%

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

et al. 2018

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“…Unfortunately, the determination of thiol compounds in biological samples still remains perplexing. The main challenge lies in their physicochemical properties [7,8]. Aside from the great susceptibility to oxidation, which can occur before or during analytical process, most thiols lack the structural properties necessary for the generation of signals compatible with common high-performance liquid chromatography (HPLC) detectors such as ultraviolet (UV) absorbance and fluorescence.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, the analyst must resort to derivatization for signal enhancement and labile sulfhydryl group blocking if fluorescence or UVvis detection methods are employed. Moreover, low concentration of biologically important thiols in real matrices, high polarity, and good solubility in water make their extraction very troublesome [7,8]. Among a variety of assays designed to determination of thiols in biological fluids, most of them depend on derivatization followed by chromatographic separation and ultraviolet [7][8][9] or fluorescence detection [10][11][12][13][14][15].…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, low concentration of biologically important thiols in real matrices, high polarity, and good solubility in water make their extraction very troublesome [7,8]. Among a variety of assays designed to determination of thiols in biological fluids, most of them depend on derivatization followed by chromatographic separation and ultraviolet [7][8][9] or fluorescence detection [10][11][12][13][14][15]. Most of chromatographic procedures dedicated to hydrophilic thiols measurements exploit reversed phase (RP)-HPLC and buffered eluents containing ion-pairing reagents, such as alkyl sulphonates or trichloroacetic acid [7][8][9][10][11][12][13][14][15][16][17][18][19][20].…”

Section: Introductionmentioning

confidence: 99%

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A simple HPLC—UV method for simultaneous determination of cysteine and cysteinylglycine in biological fluids

Głowacki¹,

Stachniuk²,

Borowczyk³

2016

Acta Chromatographica

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Summary.A new and simple method based on high-performance liquid chromatography with ultraviolet detection (HPLC-UV) for the determination of cysteine (Cys) and cysteinylglycine (CysGly) in plasma and urine has been developed. The method involves reduction of disulfide bonds with tris(2-carboxyethyl)phosphine, derivatization of the analytes with 2-chloro-1-methylquinolinium tetrafluoroborate, and separation on Aeris PEPTIDE XB-C18 column (150 mm × 4.6 mm, 3.6 µm, Phenomenex) with UV detection at 355 nm. The calibration lines, obtained with human plasma and urine spiked with CysGly and Cys, were linear in the range of 2.5-50 μmol L −1 and 20-300 μmol L −1 , respectively. The intra-and inter-day precision values of the method, expressed as a relative standard deviation, were 0.25-11.1% and 0.71-12.3%, respectively. The analytical recovery varied from 89.7 to 112.3%. The LOQs for total Cys and CysGly were 1.5 pmol and 2.3 pmol in peak, respectively. The method was successfully applied to samples donated by apparently healthy individuals. Concentrations of Cys and CysGly in human plasma from 18 subjects varied from 141.6 to 217.8 μmol L −1 and from 21.1 to 50.9 μmol L −1 , respectively. Their concentrations in urine samples (n = 14) ranged from 137.3 to 426.8 μmol L −1 and from 1.6 to 4.9 μmol L −1 , respectively.

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“…Derivatization temperature plays an important role in a derivatization reaction. Lower temperature leads to lower reaction rate and longer reaction time while higher temperature may cause the decomposition of derivatization reagents or analytes [17]. Different derivatization temperatures of 10 °C, 20 °C, 30 °C, 45 °C, 60 °C, and 75 °C were compared, and results were shown in Fig.…”

Section: Derivatization Conditionsmentioning

confidence: 99%

Determination of cembrenediols in tobacco by gas chromatography—mass spectrometry-selected ion monitoring with precolumn derivatization

Zhou

Yang

et al. 2016

Acta Chromatographica

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Summary. An efficient and sensitive analytical method based on precolumn derivatization and gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM) was proposed and validated for analysis of two cembrenediols (CBDs) which are α-cembrenediol and β-cembrenediol in tobacco samples. CBDs in tobacco samples were extracted by sonication with 50 mL dichloromethane for 10 min before derivatized with 2:3 (v/v) bis(trimethylsilyl)trifluoroacetamide (BSTFA)-pyridine at 20 °C for 100 min. CBDs' level in tobacco samples was analyzed by GC-MS-SIM and quantified by the internal standard method. The linear range for α-CBD and β-CBD was 13.6-554.6 μg mL −1 and 4.11-162.6 μg mL −1 , and the correlation coefficients of both were 0.9998. The limit of detection (LOD) and limit of quantification (LOQ) of α-cembrenediol and β-cembrenediol were 0.40 μg g −1 and 1.34 μg g −1 , and 0.27 μg g −1 and 0.90 μg g −1 , respectively. Average recoveries of α-CBD and β-CBD were 94.4-99.9% and 91.9-98.2% while the relative standard deviations (RSDs, n = 5) were ranged from 2.67 to 5.6% and 2.04 to 4.22%, respectively. This proposed analytical method has been successfully applied to analyze CBDs in tobacco samples.

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Ultraviolet derivatization of low-molecular-mass thiols for high performance liquid chromatography and capillary electrophoresis analysis

Cited by 111 publications

References 148 publications

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

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

A simple HPLC—UV method for simultaneous determination of cysteine and cysteinylglycine in biological fluids

Determination of cembrenediols in tobacco by gas chromatography—mass spectrometry-selected ion monitoring with precolumn derivatization

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