Determination of endogenous thiols and thiol drugs in urine by HPLC with ultraviolet detection

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

doi:10.1016/j.jchromb.2009.03.038

Cited by 123 publications

(90 citation statements)

References 104 publications

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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%

See 2 more Smart Citations

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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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

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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“…There have been numerous studies aimed at characterization of the redox states of cysteines in proteins owing to the inherent importance of cysteine-mediated chemistry in countless biological processes. Methods such as UV absorption spectroscopy [2], fluorescent labeling [3], and X-ray absorption spectroscopy [4] have been used to quantify or characterize the cysteine content of proteins. Characterizing cysteine content by mass spectrometry has also become a popular option as a consequence of the development of wellestablished bottom-up proteomics approaches [5][6][7] often in combination with various clever cysteine-selective derivatization methods [8].…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Cysteine Content in Proteins Utilizing Cysteine Selenylation with 266 nm Ultraviolet Photodissociation (UVPD)

Parker

Brodbelt

2016

J. Am. Soc. Mass Spectrom.

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Abstract. Characterization of the cysteine content of proteins is a key aspect of proteomics. By defining both the total number of cysteines and their bound/ unbound state, the number of candidate proteins considered in database searches is significantly constrained. Herein we present a methodology that utilizes 266 nm UVPD to count the number of free and bound cysteines in intact proteins. In order to attain this goal, proteins were derivatized with N-(phenylseleno)phthalimide (NPSP) to install a selectively cleavable Se-S bond upon 266 UVPD. The number of Se-S bonds cleaved upon UVPD, a process that releases SePh moieties, corresponds to the number of cysteine residues per protein.

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“…Apart from the several review articles on the analysis of thiols [1][2][3][4][5] and the many research articles that appear in the literature, the analytical challenge can be even pointed out by a very recent special issue of Journal of Chromatography B (Elsevier) under the characteristic title "Analysis of thiols" (Volume 877, Issue 28, 2009). …”

Section: Introductionmentioning

confidence: 99%

Automated Determination of Pharmaceutically and Biologically Active Thiols by Sequential Injection Analysis: A Review

Tzanavaras

2010

TOCBMJ

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Abstract:The present review article presents and discusses automated sequential injection (SI) based methods for the determination of pharmaceutically and biologically important thiols, namely cysteine, N-acetylcysteine, penicillamine, glutathione and captopril. The review intends to cover a range of ten years of published research on this topic (2000 -2009). The methods are classified according to the detection systems in three categories, namely spectrophotometric, fluorimetric and electroanalytical. The determination principles of the methods are discussed and the main analytical figures of merit are tabulated.

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Determination of endogenous thiols and thiol drugs in urine by HPLC with ultraviolet detection

Cited by 123 publications

References 104 publications

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

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

Characterization of the Cysteine Content in Proteins Utilizing Cysteine Selenylation with 266 nm Ultraviolet Photodissociation (UVPD)

Automated Determination of Pharmaceutically and Biologically Active Thiols by Sequential Injection Analysis: A Review

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