Determination of glutathione, cysteine and N-acetylcysteine in rabbit eye tissues using high-performance liquid chromatography and post-column derivatization with 5,5′-dithiobis(2-nitrobenzoic acid)

Nozal, Marı́a J.; Bernal, José; Toribio, Laura; Marinero, P.; Moral, O.; Manzanas, L; Rodriguez, E.

doi:10.1016/s0021-9673(97)00473-1

Cited by 45 publications

(17 citation statements)

References 29 publications

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“…One of the most important roles of the GSH antioxidant system is the detoxification of endogenously generated peroxides. GSH production is considered the first line of defense against oxidative damage and free radical generation, where GSH functions as a scavenger and co-factor in metabolic detoxification (Nozal et al, 1997). GSH levels increased in mice treated orally for 28 consecutive days with p-synephrine (30 and 300 mg/kg) and with C. aurantium extracts (4000 mg/kg corresponding to 300 mg/kg of p-synephrine respectively) (Arbo et al, 2009a).…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, the results of our study showed a significant decrease in male rat GSH status, whereas no alterations were seen in their female counterparts. This fact could be explained as an organism response to oxidative damage, shown by MDA levels, as the function of GSH is to defend against oxidative damage and free radical generation (Nozal et al, 1997).…”

Section: Resultsmentioning

confidence: 99%

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Gender differences in biochemical markers and oxidative stress of rats after 28 days oral exposure to a mixture used for weight loss containing p-synephrine, ephedrine, salicin, and caffeine

Schmitt

Arbo

Lorensi

et al. 2016

Braz. J. Pharm. Sci.

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The association of p-synephrine, ephedrine, salicin, and caffeine in dietary supplements and weight loss products is very common worldwide, even though ephedrine has been prohibited in many countries. The aim of this study was to evaluate a 28-day oral exposure toxicity profile of p-synephrine, ephedrine, salicin, and caffeine mixture (10:4:6:80 w/w respectively) in male and female Wistar rats. Body weight and signs of toxicity, morbidity, and mortality were observed daily. After 28 days, animals were euthanized and blood collected for hematological, biochemical, and oxidative stress evaluation. No clinical signs of toxicity, significant weight loss or deaths occurred, nor were there any significant alterations in hematological parameters. Biochemical and oxidative stress biomarkers showed lipid peroxidation, and hepatic and renal damage (p < 0.05; ANOVA/Bonferroni) in male rats (100 and 150 mg/kg) and a reduction (p < 0.05; ANOVA/Bonferroni) in glutathione (GSH) levels in all male groups. Female groups displayed no indications of oxidative stress or biochemical alterations. The different toxicity profile displayed by male and female rats suggests a hormonal influence on mixture effects. Results demonstrated that the tested mixture can alter oxidative status and promote renal and hepatic damages.Uniterms: p-Synephrine/toxicity profile. Ephedrine/toxicity profile. Salicin/toxicity profile. Caffeine/ toxicity profile. Dietary supplements/evaluation. Weight loss products/evaluation.A associação de p-sinefrina, efedrina, salicina, e cafeína em suplementos alimentares e produtos para perda de peso é muito utilizada em todo o mundo, embora a efedrina tenha sido proibida em muitos países. O objetivo deste estudo foi avaliar o perfil de toxicidade à exposição oral de 28 dias à associação de p-sinefrina, efedrina, salicina e cafeína (na proporção de 10:4:6:80 m/m respectivamente) em ratos Wistar machos e fêmeas. Diariamente, os animais foram observados quanto ao peso corporal, sinais de toxicidade, morbidade e mortalidade. Após 28 dias, os animais foram sacrificados e o sangue coletado para avaliações hematológicas, bioquímicas e de estresse oxidativo. Não se observaram sinais clínicos de toxicidade, tampouco perda significativa de peso, mortes, ou quaisquer alterações significativas nos parâmetros hematológicos. Biomarcadores do estresse oxidativo e bioquímicos mostraram peroxidação lipídica, danos renais e hepáticos (p < 0,05; ANOVA/Bonferroni) em ratos machos (100 e 150 mg/kg) e a redução (p < 0,05; ANOVA/Bonferroni) nos níveis de glutationa reduzida (GSH) em todos os grupos de machos tratados. Nas fêmeas, não houve indícios de estresse oxidativo, nem alterações bioquímicas. O diferente perfil de toxicidade entre os gêneros sugere influência hormonal nos efeitos de mistura administrada. A associação testada pode alterar o estado oxidativo e promover danos renais e hepáticos. Unitermos INTRODUCTIONOverweight and obesity are highly prevalent worldwide. Many consumers are seeking strategies for reducing weight to healt...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Gender differences in biochemical markers and oxidative stress of rats after 28 days oral exposure to a mixture used for weight loss containing p-synephrine, ephedrine, salicin, and caffeine

Schmitt

Arbo

Lorensi

et al. 2016

Braz. J. Pharm. Sci.

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“…A range of methods have been developed for determination of thiols, including electrochemical methods such as cathodic stripping voltammetry and polarography [37][38][39][40][41][42][43], capillary electrophoresis (CE) equipped with electrochemical detection [44], laser-induced fluorescence detection [45] or photodiode array [46,47], and HPLC with UV/vis detection [48,49], electrochemical detection [50][51][52], fluorescence detections [3,6,28,[53][54][55][56][57][58][59][60][61][62][63][64][65][66][67] or inductively coupled plasma-mass spectrometry (ICP-MS) and electrospray-mass spectrometry (ES-MS) [68][69][70][71][72][73][74][75][76][77][78][79][80]. These methods were selective and ...…”

Section: Introductionmentioning

confidence: 99%

Determination and characterization of cysteine, glutathione and phytochelatins (PC2–6) in Lolium perenne L. exposed to Cd stress under ambient and elevated carbon dioxide using HPLC with fluorescence detection

Tang

Jia

et al. 2011

Journal of Chromatography B

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“…It acts as a free radical scavenger and participates in drug detoxification and amino acid transport in the γ-glutamyl cycle (Nozal et al, 1997). Free radicals and reactive oxygen/nitrogen species in excess lead to the oxidative damage of macromolecules (Kozer et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

Quantification of reduced glutathione by HPLC‐UV in erythrocytes of hemodialysis patients

Garcia

Schott

Charão

et al. 2008

Biomedical Chromatography

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Reduced glutathione (GSH) is a well-known multifunctional antioxidant. Its depletion is linked to a number of pathologies, such as renal insufficiency. Feasible methodologies in clinical chemistry are vital. Therefore a methodology for GSH quantification was optimized and validated by HPLC-UV. Important aspects such as acid deproteinization and GSH stability were established. The erythrocytes were hemolyzed, deproteinized, derivatized with 5,5-dithio-bis (2-nitrobenzoic) acid and analyzed using HPLC, on an RP18 gradient elution, lambda=330 nm. The method was applied to hemodialysis patients (n=75) compared with healthy subjects (n=40). The assay was linear from 0.5 to 3.0 mm (r2>0.99). The intra- and inter-run reproducibilities were obtained with CV%<10%. The accuracy (bias %) ranged from 1.32 to -6.38%, and the recovery was >94%. The derivatized sample was stable for 60 days at -20 degrees C. The GSH levels in hemodialysis patients showed a significant increase compared with healthy subjects (p<0.05) and an inverse correlation with age (r=-0.286; p=0.013) was found. This method used UV detection, reduction of the phosphate concentration in the mobile phase and effective protein removal with trichloroacetic acid. The method proved to be reproducible, precise, accurate and stable. Thus, it can be suggested for routine laboratory tests for the verification of physiopathological conditions.

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Determination of glutathione, cysteine and N-acetylcysteine in rabbit eye tissues using high-performance liquid chromatography and post-column derivatization with 5,5′-dithiobis(2-nitrobenzoic acid)

Cited by 45 publications

References 29 publications

Gender differences in biochemical markers and oxidative stress of rats after 28 days oral exposure to a mixture used for weight loss containing p-synephrine, ephedrine, salicin, and caffeine

Gender differences in biochemical markers and oxidative stress of rats after 28 days oral exposure to a mixture used for weight loss containing p-synephrine, ephedrine, salicin, and caffeine

Determination and characterization of cysteine, glutathione and phytochelatins (PC2–6) in Lolium perenne L. exposed to Cd stress under ambient and elevated carbon dioxide using HPLC with fluorescence detection

Quantification of reduced glutathione by HPLC‐UV in erythrocytes of hemodialysis patients

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