An internal standard method for the unattended high-performance liquid chromatographic analysis of ascorbic acid in blood components

Kutnink, Mark A.; Hawkes, Wayne Chris; Schaus, Ellen E.; Omaye, Stanley T.

doi:10.1016/0003-2697(87)90594-x

Cited by 163 publications

(73 citation statements)

References 8 publications

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“…Shimadzu Scientific 4.2 Class-VP software was used to integrate peak areas. Ascorbate and urate were measured by paired-ion reversed-phase HPLC coupled with electrochemical detection (28).…”

Section: Methodsmentioning

confidence: 99%

Increased atherosclerosis in hyperlipidemic mice deficient in α-tocopherol transfer protein and vitamin E

Terasawa

Ladha

Leonard

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

224

156

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Although lipid peroxidation in the subendothelial space has been hypothesized to play a central role in atherogenesis, the role of vitamin E in preventing lipid peroxidation and lesion development remains uncertain. Here we show that in atherosclerosis-susceptible apolipoprotein E knockout mice, vitamin E deficiency caused by disruption of the ␣-tocopherol transfer protein gene (Ttpa) increased the severity of atherosclerotic lesions in the proximal aorta. The increase was associated with increased levels of isoprostanes, a marker of lipid peroxidation, in aortic tissue. These results show that vitamin E deficiency promotes atherosclerosis in a susceptible setting and support the hypothesis that lipid peroxidation contributes to lesion development. Ttpa ؊/؊ mice are a genetic model of vitamin E deficiency and should be valuable for studying other diseases in which oxidative stress is thought to play a role.antioxidants O xidative modification of lipoproteins (e.g., low-density lipoproteins) has been hypothesized to play a key role in the pathogenesis of atherosclerosis (1, 2). Because vitamin E is the most potent lipid-soluble antioxidant normally found on lipoproteins in the plasma, there is strong interest in the relationship between vitamin E levels and the development of atherosclerosis. In animal models and human clinical trials, studies of the effects of vitamin E supplementation on atherosclerosis have yielded conf licting results (3-8), and little is known about the effects of vitamin E deficiency on atherosclerosis development (9).The major form of vitamin E in human plasma and tissues is ␣-tocopherol (10). ␣-Tocopherol enrichment of plasma and tissues is mediated by the ␣-tocopherol transfer protein (␣-TTP), a cytosolic lipid-transfer protein expressed in the liver (11)(12)(13)(14). Although the mechanism is unknown (15), ␣-TTP is believed to selectively transfer ␣-tocopherol from lipoproteins taken up by hepatocytes via the endocytic pathway to newly secreted lipoproteins, which facilitate its delivery to peripheral tissues (12). Humans with ␣-TTP gene defects have extremely low plasma ␣-tocopherol concentrations and develop severe neurodegenerative disease unless they are treated with high doses of vitamin E (16-18).To investigate the relationship between vitamin E deficiency and atherosclerosis, we used gene targeting to disrupt the mouse ␣-TTP gene (Ttpa) and generate a genetic model of vitamin E deficiency. We then crossed the ␣-TTP-deficient mice (Ttpa Ϫ/Ϫ ) mice with apolipoprotein (apo) E knockout (Apoe Ϫ/Ϫ ) mice (19), which spontaneously develop atherosclerosis on a chow diet (20,21). This enabled us to generate Apoe Ϫ/Ϫ mice with different Ttpa genotypes (ϩ͞ϩ, ϩ͞Ϫ, and Ϫ͞Ϫ) to test the hypothesis that ␣-TTP and vitamin E deficiency increase atherosclerosis in a susceptible setting. Materials and MethodsGeneration of ␣-TTP Knockout Mice. A 14-kb 129͞Sv genomic clone containing the Ttpa gene was isolated and subcloned into pBSSKII. A sequence replacement vector was constructed by PCR amplifica...

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Section: Methodsmentioning

confidence: 99%

Increased atherosclerosis in hyperlipidemic mice deficient in α-tocopherol transfer protein and vitamin E

Terasawa

Ladha

Leonard

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

224

156

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“…DG samples were homogenized in metaphosphoric acid 10% (w/v) according to Kutnink et al (1987). Commercially available AH − (Sigma, St Louis, USA) was used as standard.…”

Section: Determination Of the Content Of Ahtmentioning

confidence: 99%

Exposure to excess dissolved iron in vivo affects oxidative status in the bivalve Mya arenaria

González

Abele

Puntarulo

2010

Comparative Biochemistry and Physiology Part C: Toxicology &

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The effect of in vivo Fe exposure on the oxidative metabolism of the bivalve Mya arenaria was studied. Fe was supplemented in natural seawater and resulted in a significant increase in the total Fe content in the bivalve digestive gland (DG) between 9 to 17 days of exposure. Mortality of treated animals increased drastically after day 18. Oxidative stress conditions were characterized in DG through assessment of the generation of reactive oxygen species (ROS) and ascorbyl radical (A•) content. Both parameters were affected following a biphasic profile showing significant increases by days 2 and 9 of Fe exposure. The content of 2-thiobarbituric acid reactive substances (TBARS) was significantly increased over control values by days 2, 9 and 17 of treatment. The labile Fe pool (LIP) in isolated DG was elevated over control values by day 7, and maintained this increase until day 17 of Fe exposure. The content of NO, assessed by EPR spin trapping, was 60% lower in DG of animals exposed for 2 days to Fe than in control values, with no further changes. The biphasic profile of oxidative stress response to Fe exposure in DG suggests that at early stages of Fe supplementation the cellular control mechanisms, such as CAT activity, were operative to limit oxidative damage, but further Fe exposure overwhelmed these abilities. Moreover, the second phase could be understood as the consequence of the exhaustion of cellular protective systems that could also involve NO.

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“…For the vitamin C analysis, 100 ml of serum was mixed with 100 ml of 5% metaphosphoric acid and centrifuged for 10 min at 3000 Â g. Then, a supernatant was taken and filtered using a 0.45 mm syringe filter. The sample of 10 ml was used for the HPLC analysis (Kutnink et al, 1987). The serum vitamin E level was measured by the Bieri method (Bieri et al, 1979).…”

Section: Serum Vitamin Measurementmentioning

confidence: 99%

Influence of maternal serum levels of vitamins C and E during the second trimester on birth weight and length

et al. 2004

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Objective: It has been known that maternal oxidative stress during pregnancy plays an important role in fetal growth. However, the association between antioxidant vitamin levels and birth outcomes is not conclusive. We investigated the relationship between maternal serum levels of vitamins C and E during the second trimester and birth weight and length. Design: Prospective cohort study. Setting: Outpatient-clinic of obstetrics, Ewha Womans University Hospital, South Korea. Subjects and methods: The study subjects were 239 healthy, pregnant women who visited an obstetric clinic for antenatal care, and their singleton live births, in Seoul, Korea, between August 2001 and March 2003. We measured the levels of vitamins C and E in maternal serum during the period 24-28 gestational weeks. Each woman was interviewed for dietary intake by trained interviewers during the second trimester. Results: The serum concentration of maternal vitamin C during the second trimester was significantly associated with birth weight and length in the group of full-term deliveries. An increase of 1 mg/ml in the serum vitamin C level increased the birth weight by 27.2 g and the birth length by 0.17 cm. When we considered the levels of vitamins C and E together in the relationship with birth weight and length, we found that the heaviest birth weight and the longest birth length belonged to the group of upper vitamin C/ upper vitamin E. However, dietary intake estimated by 24-h recall method was not a predictor of the levels of serum vitamins C and E. Conclusion: We found that maternal serum vitamin C levels during the second trimester were positively correlated with birth weight and length in full-term babies. We also found that birth weight and length were highest when the levels of both vitamins C and E were high. Our results indicate the importance of antioxidant nutrient balance for pregnant women who are exposed to various oxidants through food, drinking water, or inhaled air.

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An internal standard method for the unattended high-performance liquid chromatographic analysis of ascorbic acid in blood components

Cited by 163 publications

References 8 publications

Increased atherosclerosis in hyperlipidemic mice deficient in α-tocopherol transfer protein and vitamin E

Increased atherosclerosis in hyperlipidemic mice deficient in α-tocopherol transfer protein and vitamin E

Exposure to excess dissolved iron in vivo affects oxidative status in the bivalve Mya arenaria

Influence of maternal serum levels of vitamins C and E during the second trimester on birth weight and length

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