Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography

Kuśmierek, Krzysztof; Głowacki, Rafał; Bald, Edward

doi:10.1007/s00216-006-0454-x

Cited by 130 publications

(88 citation statements)

References 20 publications

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“…Average total CysGly content in plasma and urine was 29.99 ± 7.98 and 2.09 ± 1.26 μmol L −1 , respectively. These results are consistent with those reported earlier [9,20,25,26], and this indicates that elaborated assay can be successfully used in large populations. It is safe for the operator and environmentally friendly, meeting the needs of the biological sample analysis.…”

Section: Real Sample Analysissupporting

confidence: 92%

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: Real Sample Analysissupporting

confidence: 92%

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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“…The integration windows for each of the scan methods are shown in Table 1, and the LODs of The absolute LODs for the thiols were in the sub-picomole range, and they were better than the LC-UV methods (11,12,33), most of the MS based detection methods (5,(34)(35)(36)(37)(38)(39), and the results via fluorescence measurement (40,41).…”

Section: Figures Of Merit For the Analytical Protocolmentioning

confidence: 99%

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Rao

Xiang

Bramanti

et al. 2010

J. Agric. Food Chem.

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A liquid chromatography method with mass spectrometric detection has been developed for the simultaneous determination of six thiols in the sulfur metabolic pathway, including cysteine (Cys), homocysteine (HCys), glutathione (GSH), cysteinyl-glycine (Cys-Gly), γ-glutamyl-cysteine (Glu-Cys), and s-adenosyl-homocysteine (AdoHcy).Tris(2-carboxyethyl) phosphine (TCEP) was used as the reducing reagent and p-hydroxymercurybenzoate (PHMB) as the derivatization reagent. Thiols were extracted from three mg of yeast using water in an ultrasonic bath. The absolute detection limits for the compounds studied were in the sub-picomole range. It was found that AdoHcy, Cys, GSH, Cys-Gly, Glu-Cys, and very little HCys were present in the selenium enriched yeast sample studied, and GSH, Glu-Cys, very little AdoHcy, Cys, and Cys-Gly were present in three bakery yeasts.

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“…1,5 Thus, pursuing an a sensitive, reliable, and convenient approach for trace analyzing L-cysteine is of great significance in biological, medical, and clinical studies and attract considerable interest in recent years. Various methods have been reported for the determination of L-cysteine, such as flow injection, 6 electrochemistry, 7-13 high-performance liquid chromatography, 14,15 and capillary zone electrophoresis.…”

Section: Introductionmentioning

confidence: 99%

Highly Sensitive and Naked Eye Dual-readout Method for ʟ-Cysteine Detection Based on the NSET of Fluorophore Functionalized Gold Nanoparticles

Fu¹,

Liu²,

Wu³

et al. 2014

Bulletin of the Korean Chemical Society

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A simple, highly sensitive and selective method based on the rhodamine B-covered gold nanoparticle with dual-readout (colorimetric and fluorometric) detection for L-cysteine is proposed. A mechanism is that citratestabilized AuNPs were modified with RB by electrostatic interaction, which enables the nanometal surface energy transfer (NSET) from the RB to the AuNPs, quenching the fluorescence. In the presence of L-cysteine, it was used as a competitor in the NSET by the strongly Au-S bonding to release RB from the Au surface and recover the fluorescence, and the red-to-purple color change quickly, which was monitored simply by the naked eye. Under the optimum conditions, the detection limit is as low as 10 nM. The method possessed the advantages of simplicity, rapidity and sensitivity at the same time. The method was also successfully applied to the determination of L-cysteine in human urine samples, and the results were satisfying.

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Analysis of urine for cysteine, cysteinylglycine, and homocysteine by high-performance liquid chromatography

Cited by 130 publications

References 20 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

Determination of Thiols in Yeast by HPLC Coupled with LTQ-Orbitrap Mass Spectrometry after Derivatization with p-(Hydroxymercuri)benzoate

Highly Sensitive and Naked Eye Dual-readout Method for ʟ-Cysteine Detection Based on the NSET of Fluorophore Functionalized Gold Nanoparticles

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