Simultaneous determination of albumin and low-molecular-mass thiols in plasma by HPLC with UV detection

Borowczyk, Kamila; Monika, Wyszczelska-Rokiel; Kubalczyk, Paweł; Głowacki, Rafał

doi:10.1016/j.jchromb.2014.12.032

Cited by 34 publications

(44 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present study, for simultaneous determination of total Cys and CysGly, tris(2-carboxyethyl)phosphine hydrochloride (TCEP), which reacts with all disulfides at room temperature, was used. Our earlier study has shown that TCEP reduces all dimer and mixed disulfides, efficiently making them accessible to derivatization reagent [9,17,19]. In the present methodology, for signal enhancement and labile sulfhydryl group protection, we exploited CMQT, a highly reactive and thiol specific UV derivatization reagent [18].…”

Section: Sample Preparationmentioning

confidence: 99%

“…The derivatization scheme takes advantage of great susceptibility of the quinolinium molecule at position 2 to nucleophilic displacement and the high nucleo-philicity of the thiol group. It has been earlier shown that the reaction is completed within 1-5 min at room temperature [9,18]. In general, derivatization of Cys and CysGly is useful to increase their hydrophobicity, detectability in UV region, as well as chemical stability.…”

Section: Sample Preparationmentioning

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%

“…2-Chloro-1-methylquinolinium tetrafluoroborate (CMQT) was previously used for the determination of biologically important thiols in different matrices including plasma [9,13], defined media [12], saliva [16], plant tissues [17], and urine [20]. All of these methods allow determination of thiols in the only one matrix.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

Głowacki¹,

Stachniuk²,

Borowczyk³

2016

Acta Chromatographica

View full text Add to dashboard Cite

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.

show abstract

Section: Sample Preparationmentioning

confidence: 99%

Section: Sample Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

View full text Add to dashboard Cite

show abstract

“…[1][2][3] It plays a crucial role in maintaining the osmotic pressure of the blood compartment, transporting hormones, binding fatty acid and buffering blood pH. 4 The concentration of serum albumin in body fluids is almost considered a reliable sign of health. The normal concentration of HSA in serum is in the range of 35 -55 g/L and is approximately 30 mg/L in urine.…”

Section: Introductionmentioning

confidence: 99%

A Near Infrared Fluorescent Probe for Sensitive Determination of Human Serum Albumin

Zeng

Zhu

et al. 2016

ANAL. SCI.

View full text Add to dashboard Cite

A fluorescent probe 1 has been successfully developed to determine human serum albumin (HSA). Probe 1 expresses a dramatic fluorescence enhancement to HSA without interference from other amino acids. Under the optimal conditions, the calibration graphs are linear over the range of 0 -13.3 μg/mL with the limit of determination of 0.61 μg/mL. Thus, this probe shows high sensitivity and selectivity to HSA.

show abstract

Application of simultaneous separation and derivatization for the determination of α‐lipoic acid in urine samples by high‐performance liquid chromatography with spectrofluorimetric detection

Borowczyk

Olejarz

Chwatko

2019

Biomedical Chromatography

View full text Add to dashboard Cite

To help to clarify therapeutic functions of lipoic acid (LA) in biochemical and clinical practice we have elaborated a fast, simple and accurate HPLC method enabling determination of LA in human urine. The proposed analytical approach includes reduction of LA with tris(2‐carboxyethyl)phosphine and simultaneous separation and derivatization of the analyte with butylamine and o‐phthaldialdehyde followed by spectrofluorimetric detection at λex = 340 nm and λem = 440 nm. The assay was performed using gradient elution and the mobile phase containing 0.0025 mol L−1 o‐phthaldialdehyde in 0.0025 mol L−1 NaOH and acetonitrile. Linearity of the detector response for LA was observed in the range of 0.3–8 μmol L−1. Limits of detection and quantification for LA in urine samples were 0.02 and 0.03 μmol L−1, respectively. The total analysis time, including sample work‐up, was <20 min. The analytical procedure was successfully applied to analysis of real urine samples delivered from six healthy volunteers who received a single 100 mg dose of LA.

show abstract

Simultaneous determination of albumin and low-molecular-mass thiols in plasma by HPLC with UV detection

Cited by 34 publications

References 38 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

A Near Infrared Fluorescent Probe for Sensitive Determination of Human Serum Albumin

Application of simultaneous separation and derivatization for the determination of α‐lipoic acid in urine samples by high‐performance liquid chromatography with spectrofluorimetric detection

Contact Info

Product

Resources

About