2012 American Society for Nutrition

Effects of vitamin C supplementation on blood pressure: a meta-analysis of randomized controlled trials

1. Stephen P Juraschek, 2. Eliseo Guallar, 3. Lawrence J Appel, and 4. Edgar R Miller III

Abstract
Background: In observational studies, increased vitamin C intake, vitamin C supplementation, and higher blood concentrations of vitamin C are associated with lower blood pressure (BP). However, evidence for blood pressurelowering effects of vitamin C in clinical trials is inconsistent. Objective: The objective was to conduct a systematic review and meta-analysis of clinical trials that examined the effects of vitamin C supplementation on BP. Design: We searched Medline, EMBASE, and Central databases from 1966 to 2011. Prespecified inclusion criteria were as follows: 1) use of a randomized controlled trial design; 2) trial reported effects on systolic BP (SBP) or diastolic BP (DBP) or both; 3) trial used oral vitamin C and concurrent control groups; and 4) trial had a minimum duration of 2 wk. BP effects were pooled by random-effects models, with trials weighted by inverse variance. Results: Twenty-nine trials met eligibility criteria for the primary analysis. The median dose was 500 mg/d, the median duration was 8 wk, and trial sizes ranged from 10 to 120 participants. The pooled changes in SBP and DBP were 3.84 mm Hg (95% CI: 5.29, 2.38 mm Hg; P < 0.01) and 1.48 mm Hg (95% CI: 2.86, 0.10 mm Hg; P = 0.04), respectively. In trials in hypertensive participants, corresponding reductions in SBP and DBP were 4.85 mm Hg (P < 0.01) and 1.67 mm Hg (P = 0.17). After the inclusion of 9 trials with imputed BP effects, BP effects were attenuated but remained significant. Conclusions: In short-term trials, vitamin C supplementation reduced SBP and DBP. Long-term trials on the effects of vitamin C supplementation on BP and clinical events are needed.

Elmer
J Endocrinol Metab 2012;2(2):66-71

An NHANES Analysis of 2005 - 2006 Data Examining the Relationship Between Diabetes Mellitus and Vitamin C Ingestion
Daniel Fischmana, Venkata Subhash Gorrepatib, c, Pramil Cheriyatha

Abstract Background: Diabetes mellitus (DM) is one of the most common diseases afflicting the United States (U.S.) population. Vitamin C (Ascorbic acid) is considered to be one of the most potent anti-oxidants present in the body. Thus, we present the results of our epidemiologic analysis of whether Vitamin C ingestion is related to the development of diabetes mellitus. Methods: For the purpose of our study, we examined National Health and Nutrition Examination Survey (NHANES) data collected between 2005 and 2006. Of 10,348 participants who had data collected during this two-year period, 4979 did not meet any exclusion criteria, and were included in our data analysis. We performed a multivariate logistic regression to find out whether plasma levels of vitamin C were correlated with the development of diabetes. Results: The unadjusted odds ratios of having diabetes in the four quartiles of Vitamin C starting from lowest to highest were 1, 0.87 (95% CI of 0.67 - 1.13), 0.62 (95% CI of 0.50 - 0.78) and 0.45 (95% CI of 0.359 - 0.557), respectively. When the analysis was adjusted for risk factors the odds ratios still showed a dose-response relation with odds ratios of 1, 0.79 (95% CI of 0.61.04), 0.58 (95% CI of 0.45-0.76) and 0.53 (95% CI of 0.43-0.63), respectively. Conclusions: Our study supports the hypothesis that higher plasma levels of Vitamin C levels are protective against the development of Diabetes Mellitus. Given the limitations of our study, a prospective, randomized study looking at Vitamin C ingestion to reach predefined serum levels is warranted to further investigate the necessary logistics of Vitamin C use in the prevention of chronic diseases such as diabetes.

European Journal of Nutrition 2012, DOI: 10.1007/s00394-012-0359-8Online First

Vitamin C-related nutrientnutrient and nutrientgene interactions that modify folate status
Mark Lucock, Zo Yates, Lyndell Boyd, Charlotte Naylor, Jeong-Hwa Choi, Xiaowei Ng, Virginia Skinner, Ron Wai, Jeremy Kho and Sa Tang, et al.

Abstract
Purpose
Folate-related nutrientnutrient and nutrientgene interactions modify disease risk; we therefore examined synergistic relationships between dietary folic acid, vitamin C and variant folate genes with respect to red cell folate status.

Methods
Two hundred and twelve subjects were examined using chemiluminescent immunoassay, PCR and food frequency questionnaire to determine red cell and serum folate, 14 folate gene polymorphisms, dietary folate (natural and synthetic) and vitamin C.

Results

When examined independently, synthetic PteGlu correlates best with red cell folate at higher levels of intake (p = 0.0102), while natural 5CH3-H4-PteGlun correlates best with red cell folate at lower levels of intake (p = 0.0035). However, dietary vitamin C and 5CH3-H4PteGlun interact synergistically to correlate with red cell folate at higher levels of intake (p = 0.0005). No interaction between dietary vitamin C and PteGlu was observed. This natural nutrientnutrient interaction may provide an alternative to synthetic PteGlu supplementation that is now linked to adverse phenomena/health outcomes. On its own, vitamin C also correlates with red cell folate (p = 0.0150) and is strongly influenced by genetic variation in TS, MTHFR and MSR, genes critical for DNA and methionine biosynthesis that underpin erythropoiesis. Similarly, dietary vitamin C and 5CH3-H4-PteGlun act synergistically to modify red cell folate status according to variation in folate genes: of note, heterozygosity for 2R3R-TS (p = 0.0181), SHMT (p = 0.0046) and all three MTHFR SNPs (p = 0.0023, 0.0015 and 0.0239 for G1793A, C677T and A1298C variants, respectively) promote a significant association with red cell folate. Again, all these genes are critical for nucleic acid biosynthesis. Folate variants with the strongest independent effect on folate status were C677T-MTHFR (p = 0.0004) and G1793A-MTHFR (p = 0.0173).

Conclusions
5CH3-H4-PteGlun assimilation and variant folate gene expression products may be critically dependent on dietary vitamin C.

Jordan Journal of Pharmaceutical Sciences, Vol 5, No 1 (2012)

Mega-dose : Is it Harmful to the Liver? Biochemical and Histological Study in Rats

Khalid Abdul-Razzak, Mohammed Yacoub, Wael Hananeh, Shaymaa Arif

Abstract
Aim: To investigate if mega-doses of vitamin C would have deleterious effects on the liver in an animal model. Methods: A mega-dose of vitamin C (1000 mg/kg/day) was administered by oral gavage to male Wistar rats for 60 days. Both biochemical and histopathological measures were undertaken. Results: The results showed that a mega-dose of vitamin C significantly elevated lipid peroxidation and transaminase activity level in addition to the significant suppression of antioxidant enzymes activities. These results were consistent with the presence of histological lesions. Conclusion: A mega-dose of vitamin C is not safe and can cause liver injury.

Intake of vegetables, fruits, carotenoids and vitamins C and E and pancreatic cancer risk in The Netherlands Cohort Study
1. Mirjam M. Heinen1,,*, 2. Bas A.J. Verhage2, 3. R. Alexandra Goldbohm3,

4. Piet A. van den Brandt2

International Journal of Cancer

Volume 130, Issue 1, pages 147158, 1 January 2012

Abstract
Epidemiological data investigating the relation between fruit and vegetable consumption and pancreatic cancer risk have shown inconsistent results so far. Most case-control studies observed an inverse association with total fruit and vegetable consumption, whereas results from most cohort studies have largely been null. We examined prospectively the relation between pancreatic cancer risk and intake of vegetables, fruits, carotenoids and vitamins C and E. The Netherlands Cohort Study consisted of 120,852 men and women who completed a questionnaire at baseline in 1986, including a validated 150-item food-frequency questionnaire. After 16.3 years of follow-up, 423 cases were available for analysis. Total vegetable and total fruit consumption were not associated with pancreatic cancer risk (highest vs. lowest quintile, multivariable-adjusted hazard rate ratio = 1.23, 95% confidence interval: 0.86-1.75 and multivariable-adjusted hazard rate ratio = 0.90, 95% confidence interval: 0.66-1.24, respectively). Also, for cooked vegetables, raw vegetables and vegetables and fruits classified into subgroups, no associations were observed. Dietary carotenoids, vitamin C and E intake and supplements containing vitamin C or E were not associated with pancreatic cancer risk. The results were not modified by sex, smoking status and body mass index. In conclusion, we observed no association between a high consumption of vegetables and fruits and pancreatic cancer risk in this large cohort study, which is in agreement with previous prospective studies. Furthermore, we observed no association between the intake of carotenoids, vitamins and vitamin supplements and pancreatic cancer risk. Pancreatic cancer is the 5th leading cause of death in Europe and 4th in the United States.1, 2 Pancreatic cancer is diagnosed most often at advanced stages and patients diagnosed with pancreatic cancer have a 5-year survival rate of 6% or less.2, 3 So far, cigarette smoking, diabetes mellitus and body fatness are identified as risk factors.46 Fruits and vegetables contain numerous substances with potential anticarcinogenic activity (including vitamins, carotenoids and Allium compounds)7 and could therefore play a role in prevention of pancreatic cancer. Potential mechanisms of action include antioxidant protection against free-radical damage to DNA, enhancing immune function and inhibiting insulin-like growth factor (IGF) binding to IGF-receptors.4, 7 In addition, short-term animal experiments suggest that beta-carotene and the vitamins C and E hinder the development of preneoplastic lesions in both rat and hamster pancreas,8 but long-term studies demonstrated this inhibiting effect only for beta-carotene and vitamin C and only in rat pancreas.8, 9

Epidemiological data have shown inconsistent results so far. Most case-control studies have observed an inverse association with total fruit and vegetable consumption.1015 Among specific subgroups of vegetables, the most consistent association has been found for cruciferous or Brassica vegetables.11, 12, 15 On the other hand, results from most cohort studies have largely been null.16 22 The results from the Multiethnic Cohort Study suggests that vegetables may afford some protection against pancreatic cancer in high-risk subgroups, namely current smokers and overweight/obese persons.18 Regarding the relation between antioxidant intake and pancreatic cancer risk, intake of vitamin C and beta-carotene have been investigated most often, showing both inverse associations11, 14, 23 and no association.15, 19, 20, 24, 25 Data on use of vitamin supplements has been very sparse;10, 15, 20, 24 of these studies only one observed an inverse association with vitamin C supplement use.15 A recent Expert Panel Report found only suggestive evidence that fruits protect against pancreatic cancer, whereas the evidence was inconclusive for vegetables and vitamin C.4 In our study, we investigated the association between pancreatic cancer risk and the overall consumption of vegetables and fruits and consumption of subgroups of vegetables and fruits in a large prospective cohort study in The Netherlands. In addition, we investigated the relation between pancreatic cancer risk and dietary carotenoids, vitamins C and E and supplements containing vitamin C and E.

Statistical analysis
Analyses were performed for total vegetable consumption, total fruit consumption, consumption of vegetables and fruits combined, cooked and raw vegetables, vegetables categorized in subgroups (Brassica vegetables, cooked and raw leafy vegetables, Allium vegetables and legumes), total fruit, citrus fruit and consumption of individual vegetables and fruits as listed in the questionnaire. Total vegetable consumption is the summed total for all vegetables mentioned in the questionnaire and in the open-ended question, excluding dried pulses. Dried pulses were considered only in the analysis of legumes. The composition of each vegetable and fruit group is given in the Appendix section. In addition, analyses were performed for tomato/vegetable juice, processed orange/grapefruit juice and other juices. Other exposure variables were the carotenoids alpha-carotene, beta-carotene, lutein + zeaxanthin, lycopene and beta-cryptoxanthin, the vitamins C and E and use of any supplementincluding multivitaminscontaining vitamin C or E. Participants were categorized according to quintile of intake of relevant food groups or nutrients (with the lowest quintile of intake regarded as the reference group), depending on the sex-specific distribution in the subcohort. For vitamin C, however, the validation study had pointed out that quintiles 2 and 3 and quintiles 4 and 5 could not be distinguished;

therefore, we reduced vitamin C intake to three categories.36 Participants were categorized as users or nonusers of supplements containing vitamin C or E. Continuous variables were constructed as well. For vegetables and fruits an increment of 25 g/day was used based on data of the pilot study. This increment corresponds to a consumption frequency of approximately once per week for cooked vegetables. Age- and sex-adjusted and multivariable-adjusted hazard rate ratios (HRs) and corresponding 95% confidence intervals (95% CIs) were estimated by using Cox proportional hazards models. The total person-years at risk, estimated from the subcohort, were used in the analyses.38 Standard errors were estimated by using a robust covariance matrix estimator to account for increased variance due to sampling from the cohort.39 All analyses were conducted for both sexes combined and separately for men and women. Based on literature, the following variables were considered as potential confounders: age, sex, smoking, body mass index (BMI), intake of energy, coffee and alcohol, total red meat consumption, level of education, nonoccupational physical activity, family history of pancreatic cancer, history of diabetes mellitus, hypertension, gall-stones, cholecystectomy and gastric ulcer. These potential confounding variables were added to the multivariable-adjusted model if they (i) were associated with the disease and with total vegetable and total fruit intake and (ii) changed the age- and sex-adjusted regression coefficients by at least 10 percent (using a backwards stepwise procedure). This resulted in a multivariable-adjusted model including age at baseline (years), sex, cigarette smoking (current smoking: yes/no; number of cigarettes smoked per day; number of years of smoking), BMI (kg/m2), intake of energy (kcal/day), coffee (number of cups/day) and alcohol (g/day), total red meat consumption (g/day), family history of pancreatic cancer (yes/no) and history of diabetes mellitus (yes/no). The independent contribution of each vegetable subgroup was assessed by an analysis in which all vegetable subgroups were included in the model simultaneously. For analysis on antioxidant intake, the independent contribution of each specific vitamin and carotenoid to the risk of pancreatic cancer was assessed by an analysis in which all these were included in the model simultaneously. In additional analyses, the HRs were adjusted for total vegetable and fruit intake. For each analysis, trends were evaluated with the Wald test by assigning participants the median value for each level of the categorical exposure variable among the subcohort members and this variable was entered as a continuous term in the Cox regression model. To permit comparison, we restricted age-adjusted analyses to subjects included in multivariable-adjusted analyses (e.g., with no missing values on confounding variables), which left 3,937 subcohort members (1,930 men and 2,007 women) and 448 exocrine pancreatic cancer cases (240 men and 208 women) for analysis on fruit consumption and on

intake of carotenoids, vitamins and vitamin supplements. For the analysis on vegetable consumption, 3,734 subcohort members and 428 exocrine pancreatic cancer cases were available. The proportional hazards assumption, which was tested using the scaled Schoenfeld residuals,40 was violated for many of the exposure variables. Because early symptoms of disease before diagnosis could have influenced the results, the early cases (diagnosed within 2 years after baseline) were excluded; this resolved our problem of assumptions being violated. Therefore, all analyses were done excluding the first 2 years of follow-up: 69 subcohort members and 25 pancreatic cancer cases were excluded for the analysis on fruits, juices, carotenoids and vitamin intake and use of vitamin supplements. For the analysis on vegetable intake, 64 subcohort members and 22 pancreatic cancer cases were excluded. In our study, the overall analyses were performed on all pancreatic cancer cases. We restricted additional analyses to MCPC cases to create a group with a higher degree of diagnostic certainty of pancreatic cancer, which was shown to be important in previous studies.41, 42 We also conducted analyses separately for current, former and never smokers to determine whether smoking modifies the association of total vegetable and total fruit intake with risk of pancreatic cancer. Also, analyses for total vegetable and fruit intake were conducted stratified by BMI level (cutoff: 25 kg/m2) to examine whether fruit and vegetable intake may differentially affect those at higher risk. In addition, interactions on a multiplicative scale of total vegetable and total fruit consumption with smoking status and BMI were tested. All analyses were performed using the STATA statistical software package (intercooled STATA, version 9; Stata Corporation, College Station, TX). All p values were based on two-sided tests and considered statistically significant if <0.05.

Discussion
In our study no association was observed between pancreatic cancer risk and consumption of vegetables, fruits and juices. We also observed no association between pancreatic cancer risk and the intake of carotenoids, vitamins and vitamin supplements. These results are in agreement with other cohort studies. Results were not modified by sex, smoking status and BMI. Inverse associations have been observed with vegetable intake in several case-control studies. The vegetable items reported in these studies were diverse, including total vegetable plus fruit,12, 13 total vegetables,11, 12, 15Brassica vegetables,11, 13, 15 dark green leafy vegetables,12Allium vegetables,12 carrots,12, 13 and raw vegetables.11, 13, 15 Also for fruit items inverse associations have been reported for several types of fruit, including total fruit,10, 11, 13 citrus fruit,13 oranges,14 and bananas.14 Cohort studies on the other hand, have mainly reported null associations.1622 Only a few prospective studies observed

protective effects of vegetables and fruits on pancreatic cancer, showing inverse associations for total fruit in Japanese men (not in women),43 cabbage consumption among Swedish women,44 vegetarian protein products, beans, lentils and peas and dried fruit intake among Adventists.45 However, two of these studies had low case numbers (<150).44, 45 The Multiethnic Cohort Study observed an increased pancreatic cancer risk for high intake of fruit; this was, however, most apparent among nonsmokers and no association has been found for citrus fruit.46 For fruit juices mixed results have been observed as well, showing inverse associations12, 43 and no associations.11, 21 The Multiethnic Cohort Study found no association between total vegetable intake and pancreatic cancer risk overall; they found, however, some protection against pancreatic cancer in high-risk subgroups (i.e., current smokers and overweight/obese persons). They observed a significantly inverse association with dark green vegetable consumption among current smokers.18 They also observed an inverse association with total vegetables in overweight/obese persons (25 kg/m2). We, as well as others,21, 47 did not observe such findings. For carotenoids and vitamins, case-control studies observed inverse associations for betacarotene,11, 23 lycopene (in men),25 vitamin C,11, 14, 23 and E;14, 23 cohort studies reported only null findings on these carotenoids and vitamin intake.19, 20 Other carotenoids, such as alpha-carotene, lutein plus zeaxanthin and beta-cryptoxanthin, have only been investigated in case-control studies so far,15, 25 showing no associations with pancreatic cancer risk. Inconsistencies could have occurred because case-control studies are prone to more biases compared to cohort studies, including recall bias; risk estimates might be either exaggerated or underestimated because dietary intake is assessed in cases after diagnosis. Also selection bias is a problem due to high and rapid fatality rates of pancreatic cancer cases. Differential misclassification of the exposure could also have occurred due to the need to use next-of-kin interviews in case-control studies because pancreatic cancer is rapidly fatal. In addition, for several studies no dietary information was available on individual vegetables or fruits16, 17, 22 and some studies had small sample sizes (n cases <150).19, 22, 44, 45 In our study, an extensive list of vegetables and fruits was assessed. On the other hand, dietary assessment is liable to error and may have resulted in misclassification of exposure. Vegetables are generally considered as food items that are not very easy to assess in FFQs, particularly if portion sizes have to be estimated. We have intended to minimize the amount of uninformative data. Subjects with incomplete or inconsistent dietary data and, specifically, those participants who appeared not to have understood how to answer the questions on

vegetable consumption, were excluded. If misclassification has occurred, we expect this to be nondifferential and risk estimates will be most likely biased towards the null value. In the NLCS validation study, the correlation coefficient between the 9-day diet record and the FFQ for total vegetable consumption was 0.38.36 This correlation is low, but comparable to the figure reported for other prospective studies.43, 44, 48 One of the reasons for the low correlation could be that our study population may have been too homogeneous regarding intake and therefore may have yielded too little contrast between highest and lowest quintile of total vegetable consumption to detect differences in pancreatic cancer risk. Because of individual preferences, however, contrast in consumption frequency of many specific vegetables as well as for fruit is much higher. Although we cannot entirely exclude the possibility that the absence of protective effects of vegetables and fruits on pancreatic cancer is due to measurement error or too little contrast in our data, this can not be unique for our study and it does not explain why especially case-control studies observed protective associations. A large European cohort studywhich also did not observe an association between pancreatic cancer risk and fruit and vegetable consumptionhas a wider range of fruit and vegetable intake compared to other prospective cohort studies, caused by inclusion of participants from Northern to Southern Europeans countries.21 However, their range of fruit and vegetable intake was comparable to ours. Strengths of our study include the possibility to further restrict the analyses to MCPC cases only42 and the large sample size. Differential follow-up is unlikely to have made a material contribution to our findings, as completeness of follow-up was high.30 The prospective design avoided recall bias and the need to use next-of-kin respondents. In conclusion, we observed no association between pancreatic cancer risk and a high consumption of vegetables and fruits in the NLCS, which is in agreement with previous prospective studies. Furthermore, we observed no association between the intake of alphacarotene, beta-carotene, lutein plus zeaxanthin, beta-cryptoxanthin, lycopene and vitamins C and E and pancreatic cancer risk.
Vitamin C in mouse and human red blood cells: An HPLC assay Hongyan Li1, Hongbin Tu, 1, , Yaohui Wang, Mark Levine, Molecular and Clinical Nutrition Section, Digestive Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-1372, USAAvailable online 19 April 2012. --------------------------------------------------------------------------------

Abstract Although vitamin C (ascorbate) is present in whole blood, measurements in red blood cells (RBCs) are problematic because of interference, instability, limited sensitivity, and sample volume requirements. We describe a new technique using HPLC with coulometric electrochemical detection for ascorbate measurement in RBCs of humans, wild-type mice, and mice unable to synthesize ascorbate. Exogenously added ascorbate was fully recovered even when endogenous RBC ascorbate was below the detection threshold (25 nM). Twenty microliters of whole blood or 10 l of packed RBCs was sufficient for assay. RBC ascorbate was stable for 24 h from whole-blood samples at 4 C. Processed, stored samples were stable for >1 month at 80 C. Unlike other tissues, ascorbate concentrations in human and mouse RBCs were linear in relation to plasma concentrations (R = 0.8 and 0.9, respectively). In healthy humans, RBC ascorbate concentrations were 957 M, corresponding to ascorbate plasma concentrations of 1590 M. Mouse data were similar. In human blood stored as if for transfusion, initial RBC ascorbate concentrations varied approximately sevenfold and decreased 50% after 6 weeks of storage under clinical conditions. With this assay, it becomes possible for the first time to characterize ascorbate function in relation to endogenous concentrations in RBCs.
Oxidative Stress in Applied Basic Research and Clinical Practice, 2012, Part 5, 373-385, DOI: 10.1007/978-1-61779-397-4_18

Antioxidants, Anorexia/Cachexia, and Oxidative Stress in Patients with Advanced-Stage Cancer

Giovanni Mantovani, Clelia Madeddu and Antonio Macci

Abstract Cancer cachexia is increasingly becoming a critical component in the comprehensive approach to cancer patients influencing morbidity, mortality, and quality of life. Therefore, its pathophysiology and the main contributing factors have been investigated with the aim of developing effective therapies. Reported findings indicate that increased production of reactive oxygen species and reduced activity of antioxidant enzymes contribute to development of anorexia and cachexia in cancer. Oxidative stress, almost always accompanying cancer cachexia, may be counteracted by effective antioxidant treatments: in this review, the most relevant recent clinical approaches addressing this target are reported. Fairly advanced clinical data on efficacy of and antioxidants in advanced cancer patients are promising, but the best way to administer and combine them with other agents, the optimal dose, and timing remain uncertain. However, because cachexia is a multifactorial syndrome, therapeutic approaches targeting a single contributing factor may be inadequate: targeting oxidative stress only one determinant is addressed, thereby limiting clinical

efficacy. Therefore, antioxidants should be included in developing a therapeutic approach for cachectic cancer patients, however, they cannot encompass all symptoms of cancer cachexia. Recent evidence seems to confirm that the treatment of cancer cachexia, a multifactorial syndrome, is more likely to yield success with a multitargeted approach. A Critical Review of Vitamin C for the Prevention of Age-Related Cognitive Decline and Alzheimer's Disease Journal Journal of Alzheimer's Disease Issue Volume 29, Number 4 / 2012 , Pages 711-726 Authors Fiona E. Harrison1 Abstract Antioxidants in the diet have long been thought to confer some level of protection against the oxidative damage that is involved in the pathology of Alzheimer's disease as well as general cognitive decline in normal aging. Nevertheless, support for this hypothesis in the literature is equivocal. In the case of vitamin C (ascorbic acid) in particular, lack of consideration of some of the specific features of vitamin C metabolism has led to studies in which classification of participants according to vitamin C status is inaccurate, and the absence of critical information precludes the drawing of appropriate conclusions. Vitamin C levels in plasma are not always reported, and estimated daily intake from food diaries may not be accurate or reflect actual plasma values. The ability to transport ingested vitamin C from the intestines into blood is limited by the saturable sodium-dependent vitamin C transporter (SVCT1) and thus very high intakes and the use of supplements are often erroneously considered to be of greater benefit that they really are. The current review documents differences among the studies in terms of vitamin C status of participants. Overall, there is a large body of evidence that maintaining healthy vitamin C levels can have a protective function against age-related cognitive decline and Alzheimer's disease, but avoiding vitamin C deficiency is likely to be more beneficial than taking supplements on top of a normal, healthy diet.

Bratisl Lek Listy. 2012;113(3):135-8.

Does vitamin C prevent the effects of high dose dexmedetomidine on rat erythrocyte deformability?
Kurtipek O, Comu FM, Ozturk L, Alkan M, Pampal K, Arslan M.

Source
Gazi University Medical University, Department of Anesthesiology and Reanimation, Ankara, Turkey.

Abstract
PURPOSE: Dexmedetomidine is an anesthetic agent frequently used for sedation at the intensive care units and during general anesthesia. The purpose of our study was to investigate whether vitamin C prevents the effect of high dose dexmedetomidine on erythrocyte deformability in rats. METHODS: The study was performed on 21 male rats, with 7 rats in each study groups and the control group. The rats in the study groups were treated with intraperitoneal dexmedetomidine (10 g/kg) and intraperitoneal dexmedetomidine plus Vitamin C (ascorbic acid) (100 mg/kg ascorbic acid administered 1 hour before administration of 10 g/kg dexmedetomidine), respectively. Intraperitoneal physiological saline was administered in the control group. Erythrocyte packs were prepared using heparinized total blood samples. Deformability measurements were done by erythrocyte suspensions in phosphate buffered saline (PBS) buffer. A constant flow filtrometer system was used to measure erythrocyte deformability and the relative resistance was calculated. RESULTS: Erythrocyte deformability was significantly higher in dexmedetomidine group than in control and vitamin C plus dexmedetomidine groups (p=0.003, p=0.013, respectively). Erythrocyte deformability indexes were found similar in the control group and in the vitamin C plus dexmedetomidine group (p=0.383). CONCLUSIONS: High dose dexmedetomidine may cause functional deterioration in blood flow and tissue perfusion with negative effects in erythrocyte deformability. Vitamin C supplementation seems to reverse those negative effects and variations in erythrocyte deformability. However, our preliminary results should be confirmed in wider serious of experimental and clinical trials (Fig. 1, Ref. 27). Water Soluble Vitamins Subcellular Biochemistry, 2012, Volume 56, 67-83, DOI: 10.1007/978-94-007-2199-9_5

Vitamin C in Sepsis
John X. Wilson and F. Wu

Abstract Bacterial bloodstream infection causes septic syndromes that range from systemic inflammatory response syndrome (SIRS) and encephalopathy to severe sepsis and septic shock. Microvascular dysfunction, comprising impaired capillary blood flow and arteriolar responsiveness, precedes multiple organ failure. Vitamin C (ascorbate) levels are low in critically ill patients. The impact of ascorbate administered orally is moderate because of its limited bioavailability. However, intravenous injection of ascorbate raises plasma and tissue concentrations of the vitamin and may decrease morbidity. In animal models of polymicrobial sepsis, intravenous ascorbate injection restores microvascular function and increases survival. The protection of capillary blood flow and

arteriolar responsiveness by ascorbate may be mediated by inhibition of oxidative stress, modulation of intracellular signaling pathways, and maintenance of homeostatic levels of nitric oxide. Ascorbate scavenges reactive oxygen species (ROS) and also inhibits the NADPH oxidase that synthesizes superoxide in microvascular endothelial cells. The resulting changes in redox-sensitive signaling pathways may diminish endothelial expression of inducible nitric oxide synthase (iNOS), tissue factor and adhesion molecules. Ascorbate also regulates nitric oxide concentration by releasing nitric oxide from adducts and by acting through tetrahydrobiopterin (BH4) to stimulate endothelial nitric oxide synthase (eNOS). Therefore, it may be possible to improve microvascular function in sepsis by using intravenous vitamin C as an adjunct therapy.
Water Soluble Vitamins Subcellular Biochemistry, 2012, Volume 56, 85-103, DOI: 10.1007/978-94-007-2199-9_6

Vitamin C Transport and Its Role in the Central Nervous System

James M. May

Abstract Vitamin C, or ascorbic acid, is important as an antioxidant and participates in numerous cellular functions. Although it circulates in plasma in micromolar concentrations, it reaches millimolar concentrations in most tissues. These high ascorbate cellular concentrations are thought to be generated and maintained by the SVCT2 (Slc23a2), a specific transporter for ascorbate. The vitamin is also readily recycled from its oxidized forms inside cells. Neurons in the central nervous system (CNS) contain some of the highest ascorbic acid concentrations of mammalian tissues. Intracellular ascorbate serves several functions in the CNS, including antioxidant protection, peptide amidation, myelin formation, synaptic potentiation, and protection against glutamate toxicity. The importance of the SVCT2 for CNS function is supported by the finding that its targeted deletion in mice causes widespread cerebral hemorrhage and death on post-natal day 1. Neuronal ascorbate content as maintained by this protein also has relevance for human disease, since ascorbate supplements decrease infarct size in ischemia-reperfusion injury models of stroke, and since ascorbate may protect neurons from the oxidant damage associated with neurodegenerative diseases such as Alzheimers, Parkinsons, and Huntingtons. The aim of this review is to assess the role of the SVCT2 in regulating neuronal ascorbate homeostasis and the extent to which ascorbate affects brain function and antioxidant defenses in the CNS. Immune Netw. 2012 February; 12(1): 1826. Published online 2012 February 29. 10.4110/in.2012.12.1.18 PMCID: PMC3329599

The Analysis of Vitamin C Concentration in Organs of Gulo-/Mice Upon Vitamin C Withdrawal

Hyemin Kim,1, Seyeon Bae,1, Yeonsil Yu,1 Yejin Kim,1 Hang-Rae Kim,1 Young-il Hwang,1 Jae Seung Kang, 1,2 and Wang Jae Lee 1 Author information Article notes Copyright and License information Go to:

Abstract
Background
Vitamin C is an essential nutrient for maintaining human life. Vitamin C insufficiency in the plasma is closely related with the development of scurvy. However, in vivo kinetics of vitamin C regarding its storage and consumption is still largely unknown.

Methods
We used Gulo-/- mice, which cannot synthesize vitamin C like human. Vitamin C level in plasma and organs from Gulo-/- mice was examined, and it compared with the level of wildtype mice during 5 weeks.

Results
The significant weight loss of Gulo-/- mice was shown at 3 weeks after vitamin C withdrawal. However, there was no differences between wild-type and vitamin C-supplemented Gulo-/mice (3.3 g/L in drinking water). The concentration of vitamin C in plasma and organs was significantly decreased at 1 week after vitamin C withdrawal. Vitamin C is preferentially deposited in adrenal gland, lymph node, lung, and brain. There were no significant changes in the numbers and CD4/CD8 ratio of splenocytes in Gulo-/- mice with vitamin C withdrawal for 4 weeks. And the architecture of spleen in Gulo-/- mice was disrupted at 5 weeks after vitamin C withdrawal.

Conclusion
The vitamin C level of Gulo-/- mice was considerably decreased from 1 week after vitamin C withdrawal. Vitamin C is preferentially stored in some organs such as brain, adrenal gland and lung.

Selective Separation of Vitamin C by Reactive Extraction

Alexandra Cristina Blaga* and Teodor Malutan Faculty of Chemical Engineering and Environmental Protection, Department of Organic, Biochemical and Food Engineering, Gheorghe Asachi Technical University, D. Mangeron 71, 700050 Iasi, Romania
J. Chem. Eng. Data, 2012, 57 (2), pp 431435

Abstract

Selective separation of vitamin C from the mixture with 2-ketogluconic acid (the main byproduct from the fermentation process) by reactive extraction using Amberlite LA-2 dissolved in dichloromethane has been analyzed. The mechanism of the interfacial reaction pointed out that, indifferent of the pH value, only one molecule of vitamin C or 2-ketogluconic acid and one of extractant react at the interface, the resulting interfacial product being a hydrophobic ammonium salt. The obtained results indicated that it is possible to separate vitamin C by modifying the aqueous phase pH.

Vaginal vitamin C tablets effective for bacterial vaginosis

1. S Holt

Volume 17, Issue 1, pages 7172, March 2012

Abstract

Petersen EE, Genet M, Caserini M, Palmieri R. Efficacy of vitamin C vaginal tablets in the treatment of bacterial vaginosis: a randomised, double blind, placebo controlled clinical trial. Arzneimittelforschung 2011; 61: 2605.

Aim
To evaluate the efficacy and safety of vaginally administered vitamin C in women with bacterial vaginosis.

Design
Randomised, double-blind, placebo-controlled trial with two parallel arms.

Setting
Outpatient departments across 15 sites in Germany.

Participants
The trial recruited 277 women, aged 18 years or older, presenting with bacterial vaginosis and at least three of the following signs: white discharge that smoothly coats the vaginal walls, pH of vaginal fluid >4.5, a fishy odour of vaginal discharge before or after addition of 10% KOH, or the presence of clue cells on microscopic examination

Intervention
Participants administered silicon-coated vitamin C (250 mg) or placebo tablets, vaginally, once daily for 6 days.

Main outcome measures

The primary endpoint was the bacterial vaginosis cure rate; defined as the recovery of all inclusion criteria

Main results
In the intention-to-treat (ITT) population, cure was achieved by 55.3% of patients with Vit. C (n=141) and by 25.7% of patients with placebo (n=136). The between-group difference was 29.6% (P<0.001).

Authors' conclusion
The results support an effective and safe use of silicon-coated vitamin C vaginal tablets in the management of bacterial

Commentary
Bacterial vaginosis affects up to one-third of post-menopausal women, often recurs and is usually treated with topical antibiotics. In terms of complementary and alternative therapies, a Cochrane review did not find probiotics to be a useful treatment. Previous studies have suggested that vaginal vitamin C may be a useful treatment, with a possible mechanism being that it increases local acidification, thereby making up for the decrease in hydrogen peroxide that often results from a reduction in the number of lactobacilli present. This study was a well-powered, well reported, double-blind, parallel group, RCT. The strengths of the study were the placebo-controlled, double-blind design, the solid criteria for the diagnosis of the disease [i.e. the presence of at least three of four signs of bacterial vaginosis (Amsel criteria)], and the presence of few weaknesses. As well as efficacy, the study also demonstrated the safety of the treatment, with few adverse events reported and little difference observed between the active and placebo groups. Based on the results of this study, vitamin C can be recommended for bacterial vaginosis, particularly for women in the first trimester of pregnancy for whom some topical antibiotics are contra-indicated. Future studies could assess whether a combination of vitamin C and topical antibiotics further increases the cure rate; head-to-head studies could also compare cure rates with conventional treatment.
Current Microbiology Volume 64, Number 5 (2012), 457-462, DOI: 10.1007/s00284-012-0094-7

Ciprofloxacin-Induced Antibacterial Activity is Reversed by Vitamin E and Vitamin C

Majed M. Masadeh, Nizar M. Mhaidat, Karem H. Alzoubi, Sayer I. Al-Azzam and Ashraf I. Shaweesh

Abstract In the present study, we investigated the possible involvement of oxidative stress in ciprofloxacininduced cytotoxicity against several reference bacteria including Pseudomonas aeruginosa ATCC 27853, Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 29213, and clinical isolate of methicillin-resistant Staphylococcus aureus (MRSA). Oxidative stress was assessed by measurement of hydrogen peroxide generation using a FACScan flow cytometer. The antibacterial activity of ciprofloxacin was assessed using the disk diffusion method and by measuring the minimum inhibitory concentration (MIC). Ciprofloxacin induced a dose-dependent antibacterial activity against all bacteria where the highest tested concentration was 100 ug/ml. Results revealed that E. coli cells were highly sensitive to ciprofloxacin (MIC = 0.21 g/mL 0.087), P. aeruginosa and S. aureus cells were intermediately sensitive (MIC = 5.40 g/mL 0.14; MIC = 3.42 g/mL 0.377, respectively), and MRSA cells were highly resistant (MIC = 16.76 g/mL 2.1). Pretreatment of E. coli cells with either vitamin E or vitamin C has significantly protected cells against ciprofloxacin-induced cytotoxicity. These results indicate the possible antagonistic properties for vitamins C or E when they are used concurrently with ciprofloxacin.
Journal of Solution Chemistry Volume 41, Number 2 (2012), 351-366, DOI: 10.1007/s10953-012-9791-x

Conformation and Thermodynamic Properties of the Binding of Vitamin C to Human Serum Albumin
Yue Zhang, Zi-Qiang Xu, Xiao-Rong Liu, Zu-De Qi, Feng-Lei Jiang and Yi Liu

Abstract The binding of vitamin C, L-ascorbic acid (AsA), with human serum albumin (HSA) was investigated by various spectroscopic techniques under simulated physiological conditions. The fluorescence quenching constants (K SV) at four different temperatures (292, 298, 304, and 310 K) were obtained. The thermodynamic parameters H and S were calculated to be 6.02 kJmol1 and 84.55 Jmol1K1 using the vant Hoff equation. Additional experiments to determine the stoichiometry (n) were carried out using isothermal titration calorimetry (ITC) and cyclic voltammetry (CV). The distance, r, between AsA and the tryptophan residues of HSA was calculated to be 3.7 nm according to Frsters non-radiation energy transfer theory. The effect of AsA on the conformation of HSA was studied by means of three dimensional fluorescence spectra and CD spectra. The results indicate that

the presence of AsA resulted in a slight change of the HSA secondary structure. The effect of common ions on the binding of AsA to HSA was also examined.

Effect of vitamins C and E on antioxidant status of breastcancer patients undergoing chemotherapy

1. 2. 3. 4. 5. 6. 7. 8. N. Suhail MSc1, N. Bilal MSc1, H. Y. Khan MSc1, S. Hasan MSc1, S. Sharma PhD2, F. Khan MS3, T. Mansoor MS FICS3, N. Banu PhD1

Journal of Clinical Pharmacy and Therapeutics

Volume 37, Issue 1, pages 2226, February 2012

Summary
1. 2. 3. 4. 5. 6. 7. Top of page Abstract What is Known and Objective Methods Results and Discussion What is New and Conclusion References
What is known and Objective: Reactive oxygen/nitrogen species generated by antineoplastic agents are prime suspects for the toxic side-effects of acute or chronic chemotherapy. The present study was undertaken to test whether vitamins C and E (VCE) supplementation protect against some of the harmful effects of commonly used anticancer drugs in breast-cancer patients. Methods: In a randomized 5-month study, the activity of various antioxidant enzymes (superoxide dismutase, catalase, glutathione-S-transferase and glutathione reductase) and the levels of malondialdehyde and reduced glutathione were measured in forty untreated breastcancer patients (stage II) and compared with those of healthy controls. The degree of DNA damage was also assessed in the peripheral lymphocytes of the patients by alkaline single cell

gel electrophoresis. The untreated patients were then randomly assigned to either treatment with chemotherapy alone (5-fluorouracil 500 mg/m2 i.v. day 1, doxorubicin 50 mg/m2 i.v. day 1 and cyclophosphamide 500 mg/m2 i.v. day 1, every 3 weeks for six cycles) or to the same chemotherapy regimen supplemented with VCE (vitamin C 500 mg tablet and vitamin E 400 mg gelatin capsule). On completion of the treatments, both the groups were studied again for the levels of the markers measured prior to treatment. Results and Discussion: The untreated group showed significantly lower levels of antioxidant enzymes (P < 0001) and reduced glutathione (P < 0001), and more extensive lipid peroxidation (P < 0001) and DNA damage than healthy controls. Similar but less pronounced patterns were observed in the patients receiving chemotherapy alone. The group of patients receiving VCE supplementation had all the marker levels moving towards normal values. Activities of superoxide dismutase, catalase, glutathione-S-transferase and glutathione reductase, and the levels of reduced glutathione were significantly increased (P < 001) while, the levels of malondialdehyde and DNA damage were significantly (P < 001) reduced in the VCE supplemented group relative to those of patients receiving chemotherapy alone as well as relative to the pretreatment levels. What is new and Conclusion: Co-administration of VCE restored antioxidant status, lowered by the presence of breast-cancer and chemotherapy. DNA damage was also reduced by VCE. The results suggest that VCE should be useful in protecting against chemotherapyrelated side-effects and a randomized control trial to evaluate the effectiveness of VCE in breast-cancer patients using clinical outcomes would be appropriate.

Determination of Trace Vitamin C by Ion-Pair HPLC with UV Detection in Calcium Gluconate and Vitamin C Compound Oral Solution
1. Liandong Hu*, 2. Li Li, 3. Zhaoliang Luo, 4. Jianxue Yang and 5. Wei Liu J Chromatographic Science (2012) 50 (2): 102-107.

Abstract
A sensitive and specific reversed-phase high-performance liquid chromatographic method with ultraviolet detection was developed for the determination of vitamin C, using tetrabutylammonium hydroxide as an ion-pair reagent in a compound oral solution containing 100 mg/mL calcium gluconate and 1.25 mg/mL vitamin C. The aqueous phase contained 0.005

mol/L tetrabutylammonium hydroxide and the mobile phase consisted of a mixture of the aqueous phasemethanol (80:20, v/v, pH 6.0 adjusted by phosphoric acid). The linearity, sensitivity and specificity, accuracy, and stability of the procedure were evaluated. The calibration curves for vitamin C were linear in the range of 10.0100.0 g/mL. The percentage coefficient of variation of the quantitative analysis of the vitamin C in the products analysis was within 5%. The method was successfully applied to determine the stability of vitamin C in the compound oral solution. It was found that the vitamin C peak was symmetrical and the column efficiency was high. The method is simple and suitable for stability testing of a low concentration of vitamin C preparation.

Dietary supplementation with vitamin E and C attenuates dexamethasone-induced glucose intolerance in rats
1. Deon B. Williams1, 2. Zhongxiao Wan1, 3. Bruce C. Frier1, 4. Rhonda C. Bell1, 5. Catherine J. Field1, and 6. David C. Wright2 American Journal of Physiology 2012, 01 Jan

Abstract
Glucocorticoid excess induces marked insulin resistance and glucose intolerance. A recent study has shown that antioxidants prevent dexamethasone (DEX)-induced insulin resistance in cultured adipocytes. The purpose of this investigation was to examine the effects of dietary vitamin E and C (Vit E/C) supplementation on DEX-induced glucose intolerance in rats. We hypothesized that feeding rats a diet supplemented with Vit E/C would improve glucose tolerance and restore insulin signaling in skeletal muscle, adipose, and liver and prevent alterations in AMPK signaling in these tissues. Male Wistar rats received either a control or Vit E/C-supplemented diet (0.5 g/kg diet each of L-ascorbate and DL-all rac-alpha-tocopherol) for 9 days prior to, and during, 5 days of daily DEX treatment (subcutaneous injections 0.8 mg/g body wt). DEX treatment resulted in increases in the glucose and insulin area under the curve (AUC) during an intraperitoneal glucose tolerance test. The glucose, but not insulin, AUC was lowered with Vit E/C supplementation. Improvements in glucose tolerance occurred independent of a restoration of PKB phosphorylation in tissues of rats stimulated with an intraperitoneal injection of insulin but were associated with increases in AMPK signaling in muscle and reductions in AMPK signaling and the expression of fatty acid oxidation enzymes in liver. There were no differences in mitochondrial enzymes in

triceps muscles between groups. This study is the first to report that dietary Vit E/C supplementation can partially prevent DEX-induced glucose intolerance in rats.

LWT - Food Science and Technology

Volume 47, Issue 2, July 2012, Pages 443449

An improved sample preparation method for quantification of ascorbic acid and dehydroascorbic acid by HPLC
Kranthi K. Chebrolua, G.K. Jayaprakashaa, Kil Sun Yooa, John L. Jifona, b,
,

Bhimanagouda S. Patil

Abstract
Ascorbic acid (AA) and dehydroascorbic acid (DHA) are reduced and oxidized forms of vitaminC, which are ubiquitously found in various fruits and vegetables. The present study has evaluated and optimized various factors responsible for AA and DHA stability in grapefruit samples. Furthermore, the optimized method was used to quantify these compounds in different fruits and vegetables. The AA stability in the samples was evaluated by extracting grapefruit juice using 1, 3 and 5 g/100 mL metaphosphoric acid (MPA) and trichloro acetic acid (TCA). The AA levels were stable in grapefruit samples extracted with 1, 3 and 5 g/100 mL MPA, whereas TCA extracts showed degradation in 48 h. Among the three reducing agents studied, tris(2-carboxy ethyl) phosphine hydrochloride (TCEP) has efficiently converted DHA at all concentrations and the samples were stable for 48 h at 2.5 mmol/L TCEP. Lower pH favored complete conversion of DHA by TCEP than dithiothreitol. Among various fruits and vegetables analyzed, the highest levels of AA (260.1 mg/100 g) were observed in guava and DHA (58.6 mg/100 g) in parsley samples. The current optimized method prevents the degradation of AA and DHA from fruit and vegetable samples stored at room temperature for two days.

Phytochemistry
Volume 75, March 2012, Pages 4149

Oxidation of dehydroascorbic acid and 2,3-diketogulonate under plant apoplastic conditions

Harriet T. Parsons1, Stephen C. Fry

Abstract
The rate of L-ascorbate catabolism in plants often correlates positively with the rate of cell expansion. The reason for this correlation is difficult to explore because of our incomplete knowledge of ascorbate catabolism pathways. These involve enzymic and/or non-enzymic oxidation to dehydroascorbic acid (DHA), which may then be hydrolysed to 2,3-diketogulonate (DKG). Both DHA and DKG were susceptible to further oxidation under conditions of pH and H2O2 concentration comparable with the plant apoplast. The kinetics of their oxidation and the identity of some of the products have been investigated here. DHA, whether added in pure form or generated in situ by ascorbate oxidation, was oxidised non-enzymically to yield, almost simultaneously, a monoanion (cyclic-oxalyl-threonate; cOxT) and a dianion (oxalylthreonate; OxT). The monoanion was resistant to periodate oxidation, showing that it was not oxalic threonic anhydride. The OxT population was shown to be an interconverting mixture of 3-OxT and 4-OxT, differing in pKa. The 3-OxT appeared to be formed earlier than 4-OxT, but the latter predominated at equilibrium. DKG was oxidised by H 2O2 to two partially characterised products, one of which was itself further oxidised by H2O2 to yield threonate. The possible occurrence of these reactions in the apoplast in vivo and the biological roles of vitaminC catabolites are discussed.

Graphical abstract
During vitaminC catabolism, H2O2 non-enzymically oxidised dehydroascorbic acid to yield cyclic-oxalyl-threonate and, simultaneously, an oxalyl-threonate (OxT) pool composed of an interconverting mixture of 3-OxT and 4-OxT. Other oxidation products were partially characterised.

Highlights
Catabolism of vitaminC non-enzymically under conditions mimicking the apoplast. Dehydroascorbic acid + H2O2 yield two interconvertible oxalyl-threonate (OxT) isomers. 3OxT and 4-OxT differ in acidity and are separable by paper electrophoresis.

Diketogulonate + H2O2 yield two partially characterised acidic products. Possible signalling or defence roles of these metabolites await exploration.

Abbreviations
cOxT, cyclic oxalyl threonate; cOxTL, cyclic oxalyl threonolactone; DHA, dehydroascorbic acid; DKG, 2,3-diketogulonate; HVPE, high-voltage paper electrophoresis;

mOG, electrophoretic mobility relative to that of orange G ( mOG = 1.0) and glucose (mOG = 0.0),
corrected for electro-endo-osmosis; OG, orange G; OxA, oxalate; OxT, oxalyl threonate; ThrO, threonate

Figures and tables from this article:

Fig. 1. Proposed structures and interconversion of ascorbate metabolites Ascorbate (top left) is oxidised nonenzymically or by ascorbate oxidase to yield dehydroascorbic acid, which is shown both in its conventional structure and as the more realistic bicyclic structure adopted in aq. soln (Kerber, 2008). DHA is further oxidised by H2O2 nonenzymically (and possibly enzymically in vivo), in a reaction that cleaves the bond marked . The product is a highly reactive intermediate proposed (Parsons et al., 2011) to be cyclic-2,3-O-oxalyl-L-threonolactone (cOxTL; centre structure), which is subject to any of the three hydrolytic reactions shown, selected stochastically. If not rapidly oxidised, DHA can be hydrolysed to diketogulonate (DKG), which is shown only in its conventional structure, although this too is likely to be hydrated at the di-keto group in aq. soln. Oxalic threonic anhydride is a hypothetical product, not found; shows where periodate would cleave this structure. The CC bond originating from C-1 and C-2 of

ascorbate is in bold; the carbon derived from C-6 of ascorbate is indicated by .

Development and Validation of a Spectrophotometric Method for Quantification of Total Glucosinolates in Cruciferous Vegetables
Cynthia M. Gallaher, Daniel D. Gallaher, and Sabrina Peterson*
J. Agric. Food Chem., 2012, 60 (6), pp 13581362

Abstract
Given their putative role in chemoprevention, validated methods are needed for quantification of total glucosinolates. Based on the colorimetric reaction of ferricyanide with 1-thioglucose, released by alkaline hydrolysis of glucosinolates, we developed a simple and sensitive method for spectrophotometric quantification of total glucosinolates in cruciferous vegetables. Lyophilized and ground vegetables are extracted with 80% boiling methanol. Extracted glucosinolates are isolated using a strong anion exchange column and then hydrolyzed with 2 N NaOH to release 1-thioglucose. Ferricyanide is added, and the decrease in absorbance is measured at 420 nm, with final values adjusted for background. Recovery of internal standard (sinigrin) was 107%. Intra- and interassay coefficients of variation were 5.4% and 15.8%, respectively. Dose response was linear with sinigrin and amount of plant material extracted (R2 0.99). Using sinigrin, the lower limit of quantification was 0.6 mg. This straightforward method may be an alternative to timeconsuming and costly chromatographic methods.

Pharmaceutical Chemistry Journal Volume 45, Number 7 (2011), 440-443,

Assay of ascorbic acid, thiamine, riboflavin, nicotinamide, and pyridoxine in hexavit by HPLC
S. Yu. Garmonov, I. A. Salakhov, G. R. Nurislamova, R. N. Ismailova, . A. Irtuganova and V. F. Sopin

Abstract
A method for the assay of ascorbic acid, thiamine, riboflavin, nicotinamide, and pyridoxine in Hexavit was developed using HPLC. The conditions for chromatographic separation of vitamin mixtures with gradient elution in acidic medium on a reverse phase Discovery RP Amide C16 column were established. This method was verified by analysis of industrial vitamin formulations.

Development and validation of a rapid HPLC method for the determination of ascorbic acid, phenylephrine, paracetamol and caffeine using a monolithic column
Petra Koblov , Hana Sklenov , Ivana Brabcov and Petr Solich
Analitycal Methods, 2012, Advance Article This article reports a fast and simple liquid chromatographic method for determination of ascorbic acid, phenylephrine, paracetamol and caffeine. Salicylic acid was used as internal standard. The analytes were successfully separated in less than 5 min by isocratic elution using monolithic column, Onyx Monolithic C18 (100 4.6 mm), with mobile phase composed of acetonitrile and phosphate buffer (pH 6.50) (10 : 90, v/v) at a flow rate of 1.0 mL min1 and 25 C, sample volume was 10 L. Detection was observed at two wavelengths 210 nm (phenylephrine, paracetamol and salicylic acid) and 235 nm (ascorbic acid and caffeine). The optimized method was applied for the determination of the analytes in pharmaceutical formulation Coldrex tablets (Smithkline Beecham Consumer Healthcare, United Kingdom) commonly used in virosis treatment.

Talanta Volume 84, Issue 3, 15 May 2011, Pages 789801

A new HPLC method for the simultaneous determination of ascorbicacid and aminothiols in human plasma and erythrocytes using electrochemical detection
Abad Khana, Muhammad I. Khana, Zafar Iqbala, Yasar Shaha, Lateef Ahmada, Shabnam Nazira, David G. Watsonb, Jamshaid Ali Khana, Fazli Nasira, Abbas Khana,
, ,

Ismaila

Abstract
A new, simple, economical and validated high-performance liquid chromatography linked with electrochemical detector (HPLCECD) method has been developed and optimized for different experimental parameters to analyze the most common monothiols and disulfide (cystine, cysteine, homocysteine, methionine, reduced (GSH) and oxidized glutathione (GSSG)) and ascorbicacid present in human plasma and erythrocytes using dopamine as internal standard (IS). Complete separation of all the targets analytes and IS at 35 C on Discovery HS C18 RP column (250 mm 4.6 mm, 5 m) was achieved using 0.05% TFA:methanol (97:3, v/v) as a mobile phase pumped at the rate of 0.6 ml min1 using electrochemical detector in DC mode at the detector potential of 900 mV. The limits of detection (3 S/N) and limits of quantification (10 S/N) of the studied compounds were evaluated using dilution method. The proposed method was validated according to standard guidelines and optimization of various experimental parameters and chromatographic conditions was carried out. The optimized and validated HPLCECD method was successfully applied for the determination of the abovementioned compounds in human plasma and erythrocytes. The method will be quite suitable for the determination of plasma and erythrocyte profile of ascorbicacid and aminothiols in oxidative stress and other basic research studies.

Vitamin C Deficiency Activates the Purine Nucleotide Cycle in Zebrafish* Jay S. Kirkwood, Katie M. Lebold, Cristobal L. Miranda, Charlotte L. Wright, Galen W. Miller, Robert L. Tanguay, Carrie L. Barton, Maret G. Traber and Jan F. Stevens,1 February 3, 2012 The Journal of Biological Chemistry, 287, 3833-3841. Capsule Background: We investigated the effects of vitamin C status on the metabolome of adult zebrafish.

Results: Levels of inosine monophosphate (IMP) and AMP deaminase (AMPD) activity were enhanced in vitamin C-deficient zebrafish. Conclusion: Vitamin C deficiency activates the purine nucleotide cycle in zebrafish. Significance: The link between vitamin C deficiency and elevated AMPD activity is relevant to metabolic diseases. Abstract Vitamin C (ascorbic acid, AA) is a cofactor for many important enzymatic reactions and a powerful antioxidant. AA provides protection against oxidative stress by acting as a scavenger of reactive oxygen species, either directly or indirectly by recycling of the lipid-soluble antioxidant, -tocopherol (vitamin E). Only a few species, including humans, guinea pigs, and zebrafish, cannot synthesize AA. Using an untargeted metabolomics approach, we examined the effects of -tocopherol and AA deficiency on the metabolic profiles of adult zebrafish. We found that AA deficiency, compared with subsequent AA repletion, led to oxidative stress (using malondialdehyde production as an index) and to major increases in the metabolites of the purine nucleotide cycle (PNC): IMP, adenylosuccinate, and AMP. The PNC acts as a temporary purine nucleotide reservoir to keep AMP levels low during times of high ATP utilization or impaired oxidative phosphorylation. The PNC promotes ATP regeneration by converting excess AMP into IMP, thereby driving forward the myokinase reaction (2ADP AMP + ATP). On the basis of this finding, we investigated the activity of AMP deaminase, the enzyme that irreversibly deaminates AMP to form IMP. We found a 47% increase in AMP deaminase activity in the AA-deficient zebrafish, complementary to the 44-fold increase in IMP concentration. These results suggest that vitamin C is crucial for the maintenance of cellular energy metabolism. - Mass Spectrometry (MS)MetabolismMetabolomicsPurineVitamin CZebrafish

Carbohydrate Polymers Volume 88, Issue 3, 15 April 2012, Pages 10871092

Size-controlled self-aggregated N-acyl chitosan nanoparticles as a vitaminC carrier

Youngjin Choa, Jun Tae Kimb,

Hyun Jin Parka

Abstract
N-acyl chitosans with various acyl chain lengths were synthesized to improve their hydrophobicity and stability in delivery system. N-acyl chitosan nanoparticles were fabricated by self-aggregation method and vitaminC was loaded into the particles. Particle sizes were ranged from 444 nm to 487 nm with various acyl chain lengths reduced to particle size 216 288 nm with vitaminC loading. VitaminC in the N-acyl chitosan nanoparticles may cross-link and pull the particle wall resulting in reducing the particle size. N-acyl chitosan nanoparticles were characterized using FTIR, zeta potential, size analyzer, scanning electron microscope. The loading efficiency of vitaminC on N-acyl chitosan nanoparticles ranged 5567% and increased the loading efficiency of vitaminC too, with N-acyl chain length. VitaminC in N-acyl chitosan nanoparticles showed the controlled release properties at pH 1.3 and pH 7.4. Release rate of vitaminC load is reduced with increasing the length of acyl side chain.

Highlights
N-acyl modified chitosan nanoparticles are synthesized and characterized. They have the satisfactory vitaminC loading and sustained release behavior. N-acyl chitosan nanoparticles also are more stable in acidic environment. These nanoparticles showed great potential as a carrier of bioactive compounds.

Fig. 1. The chemical structures of (a) N-acyl chitosans and (b) vitamin C.

Journal of Analytical Chemistry Volume 67, Number 1 (2012), 14-20,

Silicon-titanium xerogels: Synthesis and application to the determination of ascorbic acid and polyphenoles
E. I. Morosanova, M. V. Belyakov and Yu. A. Zolotov

Abstract

Titanium and silicon-titanium xerogels have been obtained and it has been demonstrated that titanium(IV) incorporated into the xerogel matrix can participate in complexation reactions with ascorbic, gallic, ferulic, and caffeic acids and also rutin, quercetin, and dihydroquercetin present in aqueous solutions. In the case of dihydroquercetin 1 min of contact has been sufficient for achieving the equilibrium; the other compounds required 7 10 min. The stability constants of titanium(IV) complexes incorporated into xerogels reduced in the order dihydroquercetin > gallic acid > ascorbic acid > quercetin > caffeic acid > rutin. The procedures have been developed for solid-phase spectrophotometric determination of ascorbic, gallic, ferulic, caffeic acid as well as rutin, quercetin, and dihydroquercetin. The accuracy of the procedure has been examined by the standard addition method. The procedures for the determination of dihydroquercetin and ascorbic acid have been applied to the analysis of pharmaceutical preparations. Biosens Bioelectron. 2011 Jul 15;26(11):4326-30. Epub 2011 Apr 22.

Electrochemical impedance spectroscopy sensor for ascorbic acid based on copper(I) catalyzed click chemistry.
Qiu S, Gao S, Liu Q, Lin Z, Qiu B, Chen G.

Source
MOE Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection Technology for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, Fujian 350002, China.

Abstract
Copper(I) species can be acquired from the reduction of copper(II) by ascorbic acid (AA) in situ, and which in turn quantitative catalyze the azides and alkynes cycloaddition reaction. In this study, propargyl-functionalized ferrocene (propargyl-functionalized Fc) has been modified on the electrode through reacting with azide terminal modified Au electrode via copper(I) catalyzed azides and alkynes cycloaddition (CuAAC) reaction. The electrochemical impedance spectroscopy (EIS) measurement has been applied to test the electron transfer resistance of the Au electrode before and after click reaction. The changes of the fractional surface coverage () with different AA concentrations are characterized. It is found that the value has a linear response to the logarithm of AA concentration in the range of 5.0 pmol/L to 1.0 nmol/L with the detection limits of 2.6 pmol/L. The sensor shows a good stability and selectivity. And it has been successfully applied to the AA detection in the real samples (urine) with satisfactory results.