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Analysis of Ascorbic Acid (AA)
Fast and Easy Methodology
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printable Application Sheet
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Notes: The function of ascorbic acid in living organisms is very complex. AA vitamin has a protective
role against the harmful effects of free radicals, and its presence is necessary for the function of
numerous enzymes. Ascorbic acid is a powerful reducing compound and the ratio of ascorbic acid and
dehydroascorbic acid could be a biomarker of oxidative-reductive processes. It was reported in the
literature1 that in dementia patients, ascorbic acid and dehydroascorbic acid levels are different than
in healthy people. In the plasma of dementia patients, very low ascorbic acid levels were found (ca.
30% of the healthy control). Ascorbic acid level is an important factor to determine the protection of
the central nervous system against free radicals.
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Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-7.5P/td>
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| Dimensions |
4.6 x 75 mm |
| Solvents |
A: DI water/10 mM amm acetate
B: 90% acetonitrile/10% DI water/10 mM amm acetate |
| Gradient |
| Time (min) |
%B |
| 2 min |
hold 100% B |
| 5.0 min |
to 50% B |
| 10 min |
hold 50% B |
| 10.1 min |
to 100% B |
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| Post time |
5 min |
| Flow rate |
0.5 mL/min. |
| Injection |
5 microL |
| Samples |
Ascorbic Acid (AA) (0.2 mg in DI water)
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| Detection |
UV 254 nm |
Discussion
A robust and rapid HPLC–UV detection method was developed for the accurate determination of ascorbic acid (AA).
This very fast method can be used for very sensitive analysis of AA in human plasma. It is known that ascorbic
acid is not stable in plasma samples and therefore it is recommended that the samples for analysis of this
vitamin be kept at -70°C or -196°C.
Also, use of this column can be used in conjunction with LC-MS for analysis of AA in cerebrospinal fluid.
1. Ascorbic acid in cerebrospinal fluid — a possible protection against free radicals in the
brain Archives of Gerontology and Geriatrics, Volume 21, Issue 1, July-August 1995, Pages 43-48 Judit
Barabás, Erzsébet Nagy, István Degrell.
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