Direct liquid chromatography method for retinol, α- and γ-tocopherols in rat plasma

Rupérez, Javier; Mach, Martin; Barbas, Coral

doi:10.1016/j.jchromb.2003.10.016

Cited by 29 publications

(25 citation statements)

References 16 publications

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“…The CR-6 detection limit in plasma samples was 10 ng/mL. In good agreement with data of Ruperez et al (2004), the recovery from plasma was close to 100%.…”

Section: Determination Of Cr-6 Concentration In Plasma and Brainsupporting

confidence: 84%

“…At either 15 mins or 4 h after CR-6 administration, animals were anesthetized and perfused through the heart with saline to remove blood from brain vessels. The method used for CR-6 extraction from biological samples and detection was an adaptation of a method for detection of vitamins by direct liquid chromatography (Ruperez et al, 2004). Briefly, plasma (50 mL) and brain tissue (200 mg) were deproteinized in acetone (150 mL for plasma and 750 mL for brain tissue) and the samples were sonicated for 10 secs (plasma) or 20 secs (brain tissue) with an ultrasound microprobe (Labsonic M; Sartorius AG, Goettingen, Germany), followed by centrifugation for 4 mins at 12,000 g (Centrifuge 5415 D; Eppendorf, Hamburg, Germany).…”

Section: Determination Of Cr-6 Concentration In Plasma and Brainmentioning

confidence: 99%

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Antioxidant CR-6 Protects against Reperfusion Injury after a Transient Episode of Focal Brain Ischemia in Rats

Pérez-Asensio

Rosa

Jiménez‐Altayó

et al. 2009

J Cereb Blood Flow Metab

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Oxidative and nitrosative stress are targets for intervention after ischemia/reperfusion. The aim of this study was to explore the effect of CR-6, a vitamin-E analogue that is antioxidant and scavenger of nitrogen-reactive species. Sprague-Dawley rats had the middle cerebral artery (MCA) occluded either for 90 mins or permanently. Cortical perfusion was continuously monitored by laser-Doppler flowmetry. CR-6 (100 mg/kg) was administered orally either at 2 and 8 h after MCA occlusion, or at 2 h only. Infarct volume, neurological deficit, and signs of reperfusion injury were evaluated. CR-6 was detected in plasma and brain by HPLC. CR-6 reduced glutathione consumption in the ischemic brain and superoxide generation in the isolated MCA. CR-6 decreased infarct volume and attenuated the neurological deficit at 1 and 7 days after ischemia/reperfusion, but not after permanent ischemia. Immediately after reperfusion, cortical blood flow values returned to their baseline ( ± 20%) in several animals, whereas others showed hyper-perfusion ( > 20% of baseline). Reactive hyperemia was associated with adverse events such as increased cortical BBB leakage, edema, protein nitrotyrosination, COX-2 expression, and neutrophil accumulation; and with a poorer outcome, and CR-6 attenuated these effects. In conclusion, oral CR-6 administration after transient ischemia protects the brain from reperfusion injury.

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“…The CR-6 detection limit in plasma samples was 10 ng/mL. In good agreement with data of Ruperez et al (2004), the recovery from plasma was close to 100%.…”

Section: Determination Of Cr-6 Concentration In Plasma and Brainsupporting

confidence: 84%

Section: Determination Of Cr-6 Concentration In Plasma and Brainmentioning

confidence: 99%

Antioxidant CR-6 Protects against Reperfusion Injury after a Transient Episode of Focal Brain Ischemia in Rats

Pérez-Asensio

Rosa

Jiménez‐Altayó

et al. 2009

J Cereb Blood Flow Metab

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“…Plasma (baseline samples from preinfusion period) concentration of retinol and α-tocopherol were determined using HPLC (Hitachi, U-2010, Tomy, EX-125, Taitec, DTU-1C Shizuoka, Japan) according the method described by Rupérez et al (2004) with slight difference in UV detection. The UV detection was performed at 325 nm excitation and 470 nm emissions for up to 6 min for retinol detection, and fluorescence was employed with excitation at 295 nm and emission at 330 nm for α-and γ-tocopherol for 6 to 13 min.…”

Section: Analysesmentioning

confidence: 99%

Responses of plasma glucose metabolism to exogenous insulin infusion in sheep-fed forage herb plantain and exposed to heat

Al-Mamun

Shibuya

Kajita

et al. 2017

Animal

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The use of herbal plants as traditional medicines has a century long history. Plantain (Plantago lanceolata L.) is a perennial herb containing bioactive components with free radical scavenging activities. An isotope dilution technique using [U-13 C]glucose was conducted to determine the effect of plantain on the responses of plasma glucose metabolism to exogenous insulin infusion in sheep. Six crossbred sheep (three wethers and three ewes; mean initial BW = 40 ± 2 kg) were fed either a mixed hay of orchardgrass ( Dactylis glomerata) and reed canarygrass ( Phalaris arundinacea) (MH-diet) or mixed hay and fresh plantain (1 : 1 ratio, dry matter basis, PL-diet) and exposed to a thermoneutral (TN, 20°C; 70% relative humidity (RH)) environment or a heat exposure (HE, 30°C; 70% RH) for 5 days using a crossover design for two 23-day periods. The isotope dilution was conducted on days 18 and 23 of the experimental period during TN and HE, respectively. Plasma concentration of α-tocopherol was greater ( P < 0.0001) for the PL-diet than the MH-diet and remained comparable between environmental treatments. Plasma glucose concentration before isotope dilution technique was reduced for sheep ( P = 0.05) during HE compared with TN and remained comparable between diets. Plasma glucose turnover rate during the preinfusion period of insulin did not differ ( P = 0.10) between dietary treatments and between environments ( P = 0.65). The response of plasma glucose utilization to exogenous insulin administration was lower ( P = 0.04) for the PL-diet than the MH-diet. Under present experimental conditions, the plantain group was found to be resistant to the effects of insulin infusion.Keywords: glucose metabolism, plantain, insulin, heat stress, sheep Implications Plantain (Plantago lanceolata L.) is a perennial herb having bioactive components. The present experiment evaluated the effect of plantain on the responses of plasma glucose metabolism to exogenous insulin in heat-exposed sheep. Plasma glucose turnover rate (TR) was affected neither by feeding plantain nor exposure to a hot environment (30°C, 70% relative humidity (RH)). However, the response of plasma glucose utilization to exogenous insulin infusion was reduced in sheep when 50% of the mixed hay of orchardgrass (Dactylis glomerata) and reed canarygrass (Phalaris arundinacea) diet was replaced by fresh plantain.

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“…Although normal phase methods have been employed for the separation of some compounds [9][10][11][12][13], the vast majority of high performance liquid chromatography based fat soluble vitamin assay methods have employed reversed phase chromatography to facilitate analyte separation . Ultraviolet detection of these compounds has been most common in the literature [42][43][44][45][46][47], but fluorescence detection has also been used in the detection of tocopherol and retinol [32][33][34][35][36][37][38][39][40][41][42]. The use of fluorescence detection has allowed for various compounds to be analyzed at low limits of quantification and detection in plasma-human [32][33][34][35][36][37]40] or otherwise [39] as an alternative to ultraviolet detection methods.…”

Section: Introductionmentioning

confidence: 99%

Fluorometric Determination of Vitamin Constituents in Human Plasma Using Ultra Performance Liquid Chromatography

Bell¹

2012

J Chromatogr Sep Tech

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Purpose: A rapid, selective and sensitive ultra performance liquid chromatography method has been developed for the detection and quantification of several vitamins in human plasma.Methods: Alpha tocopherol, gamma tocopherol, and retinol are assayed using fluorescence detection. Excitation/emission wavelengths are 295 nm/330 nm and 325 nm/470 nm for the analysis of both tocopherols and retinol, respectively. Retinol acetate is employed as the internal standard. The reversed phase method incorporates gradient elution with a mobile phase consisting of methanol and acetonitrile. Separation of vitamin compounds is achieved using a bridged ethyl-hybrid C18 column. Results:The retention times for retinol, retinol acetate, gamma tocopherol and alpha tocopherol are 1.6 minutes, 1.8 minutes, 3.9 minutes and 4.3 minutes, respectively. The limits of quantification for retinol, gamma tocopherol and alpha tocopherol, were 0.02 μg/ml, 0.02 μg/ml, and 0.1 μg/ml, respectively. Conclusion:The assay method is suitable for the analysis of these vitamins in human plasma following the ingestion of foods fortified with these fat soluble vitamins.

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Direct liquid chromatography method for retinol, α- and γ-tocopherols in rat plasma

Cited by 29 publications

References 16 publications

Antioxidant CR-6 Protects against Reperfusion Injury after a Transient Episode of Focal Brain Ischemia in Rats

Antioxidant CR-6 Protects against Reperfusion Injury after a Transient Episode of Focal Brain Ischemia in Rats

Responses of plasma glucose metabolism to exogenous insulin infusion in sheep-fed forage herb plantain and exposed to heat

Fluorometric Determination of Vitamin Constituents in Human Plasma Using Ultra Performance Liquid Chromatography

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