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Antioxidants in Human Urine
Trans-3-hydroxycinnamic acid (3-HCA) known antioxidant analyzed
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printable Application Sheet
Figure B: EIC – extracted ion chromatogram of selected compound and corresponding spectrum.
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Notes: Hydroxycinnamic acids exhibit antioxidative activity and are present in plants. It is important to determine the
antioxidant capacity of plant extracts by measuring the concentration levels of hydroxicinnamic acids before and after
enzymatic treatments. The determination of the concentration of 3HCA is essential when determining protective effects
against free radical-induced damage.
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Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-15P-2 |
| Dimensions |
2.1 x 150 mm |
| Solvents |
A: DI water + 0.1% formic acid
B: acetonitrile + 0.1% formic acid |
| Mobile Phase |
Gradient t0 = 1.44 min
| Time |
%B |
Time |
%B |
| 0.0 |
95 |
13.1 |
50 |
| 0.2 |
95 |
14.0 |
50 |
| 12.0 |
75 |
14.1 |
95 |
| 13.0 |
75 |
20.0 |
95 |
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| Flow Rate |
0.4 mL/min. |
| Sample |
Human urine - after simple extraction |
| Peak |
Trans-3-hydroxycinnamic acid 165.05405
m/z (M+H)+, RT = 10.92 min |
| Detection |
ESI – pos - Agilent 6210 MSD TOF mass spectrometer |
Discussion
Polar metabolites are reproducibly retained using an MS friendly, ANP (inverse gradient) mobile phase. The sample preparation
required is very easy and cost effective ($3 per sample to $0.35 per sample of urine) compared to ordinary methods. The sample is
diluted with the ANP solvents and ready to inject in 5-10 minutes. With this method, investigating metabolite excretion of laboratory
animals or determining endogenous urinary biomarkers or working on phenolic composition of plant extracts is simple and easy.
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