Determination of biologically active low-molecular-mass thiols in human blood

Ivanov, A.R; Nazimov, Igor V.; Baratova, L. A.

doi:10.1016/s0021-9673(00)00713-5

Cited by 111 publications

(39 citation statements)

References 30 publications

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“…Because of its nearly ideal fluorescence properties, BODIPY is the best choice of fluorophore for a fluorescent labeling probe. Most BODIPY-based labeling probes in HPLC fluoresce around 500 nm [9][10][11][12][13][14][15], which has substantial spectral overlap with auto-fluorescence from biological samples. BODIPY can be easily modified to redshift its wavelengths by expanding its conjugated system, and we believe it could be useful to substitute the 3 and 5-methyl groups of 1,3,5,7-tetramethyl-BODIPY (see Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Because most thiol compounds have no fluorescence, an excellent fluorescent labeling probe is desired for use in HPLC-FD. Many fluorescent probes for determination of thiols with HPLC have been reported, including monobromobimane (mBrB) [9,10], 7-fluorobenzo-2-oxa-1,3-diazoloe-4-sulfonamide (SBD-F) dual pumps, CTO-10 column oven, and RF-10AXL detector was used. Separations were performed on a Kromasil C 18 column (150 ×4.6 mm, i.d.…”

Section: Introductionmentioning

confidence: 99%

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A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols

Zhang

Guo

et al. 2014

Anal Bioanal Chem

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A new long-wavelength fluorescent probe, 1,7-dimethyl-3,5-distyryl-8-phenyl-(4'-iodoacetamido)difluoroboradiaza-s-indacene (DMDSPAB-I), was designed and synthesized for thiol labeling in high-performance liquid chromatography (HPLC). The excitation and emission wavelengths of DMDSPAB-I are 620 and 630 nm, respectively, with a high fluorescence quantum yield of 0.557, which is advantageous in preventing interference of intrinsic fluorescence from complex biological matrices and enabling high sensitivity HPLC. Based on DMDSPAB-I, a reversed-phase HPLC method was developed for measuring low-molecular-weight thiols including glutathione, cysteine, homocysteine, N-acetylcysteine, cysteinylglycine, and penicillamine. After the specific reaction of DMDSPAB-I with thiols, baseline separation of all six stable derivatives was achieved through isocratic elution on a C18 column within 25 min, with the limits of detection (signal-to-noise ratio = 3) from 0.24 nmol L(-1) for glutathione to 0.72 nmol L(-1) for penicillamine. The proposed method was validated in part by measuring thiols in blood samples from mice, with recoveries of 95.3-104.3%.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols

Zhang

Guo

et al. 2014

Anal Bioanal Chem

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“…Para a determinação do GSSG, foi seguido o procedimento descrito por Ivanov e cols. (17) que utiliza uma técnica de HPLC com detecção fluorescente prévia de tiois livres, com trifenilfosfina e conjugação posterior com monobromobimano que detecta quantidades de aminotiois de 2 pmol. As extrações de sangue foram feitas, de ambos os grupos, antes do início do programa de atividade física (Medida 1) e ao final da intervenção (Medida 2), por profissionais sanitários especializados.…”

Section: Parâmetros E Indicadores Avaliadosunclassified

Antioxidantes, atividade física e estresse oxidativo em mulheres idosas

Rebelatto

Jiménez

Delgado³

et al. 2008

Rev Bras Med Esporte

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Objetivo: Verificar a influência da suplementação de vitaminas antioxidantes na dieta de mulheres idosas que praticam exercícios físicos regulares, sobre o estresse oxidativo, indicadores da saúde física e risco de enfermidades cardiovasculares (ECV). Método: Foram observados dois grupos (S e C) de mulheres com idades entre 60 e 80 anos participantes de um programa de atividades físicas durante 58 semanas, com freqüência de três vezes por semana e duração de 50 a 55 minutos cada sessão. A dieta habitual do Grupo S (n=36) foi suplementada diariamente com 330 ml de uma bebida antioxidante (Funciona TM ); o Grupo C (n=32) ingeriu água e se caracterizou como controle. Como indicadores do estresse oxidativo foram determinadas as concentrações plasmáticas de glutationa reduzida (GSH) e oxidada (GSSG), calculada a relação molar GSH/GSSG, e identificado o dano oxidativo em lipídios e proteínas. As condições físicas e cardiovasculares foram avaliadas por meio dos parâmetros antropométricos habituais (peso, altura e índice de massa corporal) e da pressão arterial. Resultados: O Grupo C apresentou aumentos significativos do estresse oxidativo, redução da pressão arterial e dos valores médios de indicadores de risco de ECV. O Grupo S teve o estresse oxidativo reduzido significativamente e apresentou incremento dos ganhos cardiovasculares. Não foram identificadas significâncias em relação aos efeitos ergogênicos. Conclusão: Os dados indicam que mulheres idosas que realizam exercícios físicos freqüentes melhoram suas condições físicas e cardiovasculares e que o suplemento dietético continuado de alimentos funcionais antioxidantes podem minimizar os efeitos danosos das espécies reativas de oxigênio.Palavras-chave: atividade física, antioxidantes, estresse oxidativo, idosos, saúde cardiovascular. ABSTRACTObjective: To verify the influence of dietary antioxidant supplementation in older women who regularly practice physical activities, on the occurrence of oxidative stress, physical health and risk of cardiovascular diseases (CVD). Method: Two groups (S and C) of women, with age ranging from 60 to 80 years old, were observed. Both groups took part in a physical activity program for 58 weeks, three times a week, for about 50 to 55 minutes each session. The diet of group S (n=36) was daily supplemented with 330 ml of a functional antioxidant beverage, Funciona TM ; Group C (n=32) ingested water and was used as Control. As oxidative stress indicators, the plasmatic concentrations of reduced (GSH) and oxidized (GSSG) glutathione were determined; the molar GSH/GSSG ratio was calculated, and the oxidative damage in lipids and proteins was evaluated. The physical and cardiovascular conditions were evaluated through routine anthropometric parameters (weight, stature and BMI) and blood pressure. Results: Group C presented significant increases of oxidative stress, reduction in the blood pressure and in the indicators of cardiovascular risks. Group S presented significant reduction of the oxidative stress and increment of the cardiova...

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“…A series of analytical procedures based on high-performance liquid chromatography (HPLC) [6][7][8], gas chromatography-mass spectrometry (GC-MS) [9], enzymatic [10,11], immunoassay [12], molecular imprinting [13], and electrochemical [14] methods have been described to be effective for HCys determination during the past decade. However, tedious analytical procedures and expensive reagents and instrumentation are necessary for such determination methods.…”

Section: Introductionmentioning

confidence: 99%

Use of capillary electrophoresis with chemiluminescence detection for sensitive determination of homocysteine

Zhao

Huang

et al. 2011

J of Separation Science

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A sensitive capillary electrophoresis (CE) method with chemiluminescence (CL) detection was developed for the determination of homocysteine (HCys) in human plasma. In this work, N-(4-aminobutyl)-N-ethylisoluminol was used as tagging reagent to label the analyte for achieving high assay sensitivity. N-(4-Aminobutyl)-N-ethylisoluminol-tagged HCys after CE separation reacted with hydrogen peroxide in the presence of horseradish peroxidase, producing CL emission. Experimental conditions for labeling analyte, CE separation, and CL detection were studied. The CL intensity was proportional to the concentration of HCys in the range of 2.5×10(-8) to 5.0×10(-6) M. Detection limit (S/N=3) was 7.6×10(-9) M. Human plasma samples from healthy donors were analyzed by the presented method. HCys levels were found to be in the range of 9.50-15.3 μM.

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Determination of biologically active low-molecular-mass thiols in human blood

Cited by 111 publications

References 30 publications

A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols

A new BODIPY-based long-wavelength fluorescent probe for chromatographic analysis of low-molecular-weight thiols

Antioxidantes, atividade física e estresse oxidativo em mulheres idosas

Use of capillary electrophoresis with chemiluminescence detection for sensitive determination of homocysteine

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