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Vitamin C in Mouse and Human Red Blood Cells: an HPLC Assay.
MedLine Citation:
PMID:  22522059     Owner:  NLM     Status:  Publisher    
Although vitamin C (ascorbate) is present in whole blood, measurements in red blood cells (RBCs) are problematic because of interferences, 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, wildtype mice, and mice unable to synthesize ascorbate. Exogenously added ascorbate was fully recovered even when endogenous RBC ascorbate was below detection threshold (25 nM). Twenty μl of whole blood or 10 μl of packed RBCs were sufficient for assay. RBC ascorbate was stable for 24 hours from whole blood samples at 4(o)C. Processed, stored samples were stable for > one month at -80°C. Unlike other tissues, ascorbate concentrations in human and mouse RBCs were linear in relation to plasma concentrations (R = 0.8, 0.9 respectively). In healthy humans, RBC ascorbate concentrations were 9-57 μM, corresponding to ascorbate plasma concentrations of 15-90 μM. Mouse data were similar. In human blood stored as if for transfusion, initial RBC ascorbate concentrations varied approximately 7 fold, 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.
Hongyan Li; Hongbin Tu; Yaohui Wang; Mark Levine
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-4-18
Journal Detail:
Title:  Analytical biochemistry     Volume:  -     ISSN:  1096-0309     ISO Abbreviation:  -     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-4-23     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Published by Elsevier Inc.
Molecular and Clinical Nutrition Section, Digestive Diseases Branch National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Building 10 Room 4D 52 MSC 1372, Bethesda MD 20892-1372.
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