Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

Solichová, Dagmar; Korecká, Lucie; Svobodová, Iveta; Musil, F; Bláha, V.; Zdánský, P; Zadák, Z

doi:10.1007/s00216-003-1886-1

Cited by 10 publications

(14 citation statements)

References 7 publications

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“…The level of a-tocopherol in the erythrocyte membrane was analysed using a modified HPLC method previously published by Solichová et al (32). A Chromolith Performance RP-18e monolithic column, 10034.6 mm (Merck) with the same sample pretreatment that was applied prior to HPLC analysis was used for the chromatographic separation.…”

Section: Methodsmentioning

confidence: 99%

The Effect of LDL-Apheresis and Rheohaemapheresis Treatment on Vitamin E

Solichová

Bláha

Aufartová

et al. 2015

J Nutr Sci Vitaminol

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A complete change in lifestyle, a strict diet and intensive combined pharmacotherapy (HMG CoA reductase inhibitors are the most important components) are necessary for patients with familial hypercholesterolemia (FH) and provide excellent results in most patients. Unfortunately, these measures are not sufficient for all patients (1-6). Homozygous FH patients usually fail to show any response to pharmacotherapy. LDL-apheresis has become the standard treatment for individuals homozygous for FH (7-9). In homozygous FH, statins provide a range of responses, decreasing the plasma LDL anywhere from 0% to 48%, with an average of approximately 15% for receptor-negative individuals and approximately 26% for receptor-defective individuals (5). LDL-apheresis could be offered to treat symptomatic coronary heart disease in individuals who are heterozygous FH and whose LDL remains high (.160 mg/ dL, 4.2 mmol/L) or decreases by ,40% in response to maximal medical treatment (1).Term LDL-apheresis was originally used in 1981 by Stoffel et al. (10) as an immunoadsorption elimination extracorporeal method (10). Nowadays, "LDL-apheresis" or "lipoprotein apheresis" is used for all types of extracorporeal eliminations of cholesterol. In our study we used a modification of the Stoffel and Borberg LDLapheresis-immunoadsorption method and also double filtration plasmapheresis (the primary filter is replaced by a centrifugal separator), which additionally eliminates fibrinogen and has an influence on rheological properties. For our study we will use for the first mentioned method the term "LDL-apheresis" and for the second mentioned method the term "rheohaemapheresis."LDL-apheresis is an invasive and relatively expensive but safe procedure (11, 12); it decreases cholesterol radically but it is also troubling to the patient's standard AS CR v.v.i., Olomouc, Czech Republic (Received June 26, 2014) Summary Lipid apheresis (extracorporeal lipoprotein elimination) is administered to patients with familial hypercholesterolemia who fail to respond to standard therapy. The nature of the treatment process raises the suspicion that it decreases not only cholesterol but also antioxidants. A group of 12 patients (average age 47617 y, 4 homozygous and 8 heterozygous individuals) with familial hypercholesterolemia treated by LDL-apheresis or rheohaemapheresis for 3-12 y was included in the study. In addition to cholesterol and triacylglycerol levels, vitamin E and vitamin A and also other markers of antioxidant activity were investigated. Nevertheless, the most important determined parameter was the vitamin E/cholesterol ratio in serum and lipoproteins. The results indicate that both extracorporeal elimination methods are effective and suitable ways to treat severe familial hypercholesterolemia, as the LDL fraction of cholesterol decreased by approximately 77% and 66% following LDL-apheresis and rheohaemapheresis, respectively. In addition, the serum vitamin E decreased by 54% and 57% and the decrease of the serum vitamin A was approximately 20%...

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Section: Methodsmentioning

confidence: 99%

The Effect of LDL-Apheresis and Rheohaemapheresis Treatment on Vitamin E

Solichová

Bláha

Aufartová

et al. 2015

J Nutr Sci Vitaminol

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“…After the separation of the plasma, the erythrocytes were washed three times with 0.9% sodium chloride solution containing 0.01% butylated hydroxyl-toluene and subsequently diluted 1 : 1 (v : v) with the same solution ( 19 ). The samples were then stored in liquid nitrogen at Ϫ 170˚C until analysis.…”

Section: Methodsmentioning

confidence: 99%

“…After centrifugation (1,900 ϫ g , 10 min, 0˚C) an aliquot (2,000 L) of the clean extract was separated and evaporated in a concentrator (AD5301, Eppendorf, Hamburg, Germany) at 45˚C. The residue was dissolved in 400 L of methanol and analysed by reversed-phase HPLC using internal standard calibration ( 19 ).…”

Section: Methodsmentioning

confidence: 99%

Erythrocyte Alpha-Tocopherol in Breast Cancer Patients Treated with Letrozol

Melichar

Plíšek

Hyšpler

et al. 2010

J Nutr Sci Vitaminol

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SummaryIn an earlier study, we have observed an increase of alpha-tocopherol in breast cancer patients treated with third-generation aromatase inhibitors that was related to tamoxifen withdrawal. We report here the results of measurement of alpha-tocopherol in erythrocytes and alpha-tocopherol/cholesterol ratios in patients treated with letrozol. Alpha-tocopherol in lipoprotein fractions and erythrocytes was determined by high-performance liquid chromatography in 124 post-menopausal women with breast cancer treated with letrozol immediately before the start of treatment as well as 2 and 4 mo later. After a transient decrease after 2 mo of letrozol therapy, erythrocyte alpha-tocopherol concentrations returned to pre-treatment levels 4 mo after the start of treatment. Apart from lower cholesterol in patients pre-treated with tamoxifen, no significant differences were observed at baseline between patients previously treated with tamoxifen and patients who had no prior tamoxifen in any of the other parameters investigated, but the transient decrease of erythrocyte alpha-tocopherol was observed only in patients previously treated with tamoxifen. Alpha-tocopherol content of lipoprotein fractions was significantly increased 4 mo after the start of therapy, but this increase was evident mostly in patients not treated earlier with tamoxifen. In conclusion, only minor changes of alpha-tocopherol, including a transient decrease of alpha-tocopherol in erythrocyte membranes, and an increase of alphatocopherol in lipoprotein fractions were observed during the first 4 mo of letrozol therapy.

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“…The method used in this study for the analysis of vitamin A (retinol) and vitamin E (α-tocopherol) in serum and lipoprotein fractions was modified from the method previously published by Urbanek et al 20 . The level of α-tocopherol in erythrocyte membranes was analysed by the modified HPLC method previously published by Solichova et al 21 .…”

Section: Analytical Determination Of Retinol and α-Tocopherol In Serumentioning

confidence: 99%

Blood levels of antioxidants during age-related macular degeneration treatment by rheohaemapheresis

Aufartová¹,

Bláha²,

Kasalová³

et al. 2015

BIOMED PAP

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Aims. Rheohaemapheresis treatment influences rheological markers and most likely improves metabolism in affected retinal areas, resulting not only in absorption of soft drusen but also reduction or complete disappearance of drusenoid retinal pigment epithelium detachments. However, the character of the treatment process has raised suspicion that there is a decrease not only in cholesterol but also in antioxidants, such as vitamin E and vitamin A. Methods. Twenty-three patients with the progressive dry form of age-related macular degeneration were each treated with 8 procedures of rheohaemapheresis. We measured levels of vitamin E (α-tocopherol), the vitamin E/cholesterol ratio in serum and lipoproteins (VLDL, LDL, HDL). Vitamin E in erythrocyte membrane and serum vitamin A (retinol) were also measured. These parameters were determined before and after rheohaemapheresis. Erythrocyte superoxide dismutase, erythrocyte glutathione peroxidase and serum malondialdehyde were analysed as markers of antioxidant activity and lipid peroxidation, respectively. Results. In serum, the VLDL and LDL fraction ratios of vitamin E/cholesterol increased significantly. Additionally, the HDL fraction ratio showed an increase but this was not statistically significant. The patients showed no clinical signs of vitamin E deficiency, and their serum concentrations of vitamin E did not differ from normal values. The results show that rheohaemapheresis in addition to causing a significant reduction in atherogenic LDL cholesterol, may have favourable additive anti-atherogenic effects due to a relative increase in the content of vitamin E in the lipoprotein fractions.

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Development and validation of HPLC method for the determination of α-tocopherol in human erythrocytes for clinical applications

Cited by 10 publications

References 7 publications

The Effect of LDL-Apheresis and Rheohaemapheresis Treatment on Vitamin E

The Effect of LDL-Apheresis and Rheohaemapheresis Treatment on Vitamin E

Erythrocyte Alpha-Tocopherol in Breast Cancer Patients Treated with Letrozol

Blood levels of antioxidants during age-related macular degeneration treatment by rheohaemapheresis

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