Fluorometric determination of total vitamin C in whole blood by high-performance liquid chromatography with pre-column derivatization.
Author(s): Speek AJ, Schrijver J, Schreurs WH
Publication: J Chromatogr, 1984, Vol. 305, Page 53-60
PubMed ID: 6707154 PubMed Review Paper? No
Suggested by: ISBER
Purpose of Paper
The purpose of this paper was to determine the stability of vitamin C and its quinoxaline derivative in whole blood specimens.
Conclusion of Paper
In whole blood, stabilized with ethyleneglycolbis-(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) and glutathione, vitamin C was stable at -20 degrees C for 8 days, but degradation was noted as early as 1 day when specimens were stored at 4 or 22 degrees C. The highly fluorescent quinoxaline derivative of vitamin C, 3-(1,2-dihydroxyethyl)furo[3,4-b] quinoxaline-l-one (DFQ), was stable in the dark at 4 or -20 degrees C for at least 1 day, but decreased rapidly when stored at 22 degrees C. The rate of decrease was greater when the specimen was stored in the light at 22 degrees instead of the dark.
Studies
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Study Purpose
The purpose of this study was to determine the stability of spiked vitamin C and its quinoxaline derivative in whole blood specimens.
Summary of Findings:
In whole blood, stabilized with EGTA and glutathione, vitamin C was stable at -20 degrees C for 8 days, but degradation was noted as early as 1 day when specimens were stored at 4 or 22 degrees C. For blood specimens, an oxidation time of at least 25 minutes was necessary to achieve maximum conversion of vitamin C to dehydro-l-ascorbic acid (DHAA) and then a reaction time of 2 minutes or more was required for conversion to DFQ. DFQ was stable in the dark at 4 or -20 degrees C for at least 1 day, but decreased rapidly when stored at 22 degrees C. The rate of decrease was greater when the specimen was stored in the light at 22 degrees instead of the dark.
Biospecimens
Preservative Types
- Other Preservative
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Small molecule HPLC Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Storage Storage temperature 4 degrees C
22 degrees C
-20 degrees C
Storage Storage duration 0 days
1 days
2 days
3 days
4 days
6 days
8 days
9 days
10 days
Storage Storage conditions In the dark
Exposed to light
Biospecimen Preservation Fixative additive/buffer EGTA-glutathione