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Histamine Retention
LC-MS Compatible Method
Click here to view
printable Application Sheet
Figures:
- Old red wine, TIC.
- Old red wine, EIC (112.0869 m/z). Extra peak is likely an isomer of histamine.
- Histamine dihydrochloride standard, EIC (112.0869 m/z). 50% isopropanol in solvent A.
- Histamine dihydrochloride standard, EIC (112.0869 m/z). No isopropanol in solvent A.
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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: | 50% DI H2O/ 50% 2-propanol/ 0.1% formic acid (Figures A–C)
DI H2O/ 0.1% formic acid (Figure D) |
| B: | Acetonitrile/ 0.1% formic acid (All Figures) |
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| Gradient |
| time (min.) |
%B |
time (min.) |
%B |
| 0 |
80 |
7 |
10 |
| 5 |
10 |
8 |
80 |
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| Post Time |
5 min (can be reduced for standards) |
| Injection Vol. |
1 microL |
| Flow rate |
0.4 mL/min. |
| Detection |
ESI – POS - Agilent 6210 MSD TOF mass spectrometer |
| Sample |
Old Red Wine: Sample was filtered with 0.45 microm nylon syringe filter (MicroSolv Tech Corp) and
diluted 1:5 with 50:50 solvent A:B mixture.
Standard: 1 mg/mL Histamine dihydrochloride stock solution in 80/20 DI H2O/MeOH diluted 1:100 with 50/50
solvent A/solvent B diluent. |
| Peak |
Histamine (112.0869 m/z) |
| t0 |
0.9 min |
Discussion
This method for red wine analysis is simple, fast, robust and does not require derivatization like many other HPLC methods. When isopropanol was
not used in the A solvent, the histamine peak was highly symmetrical in the standard (Compare Fig. C & D) but distorted in the red wine extract
due to matrix effects. Addition of 50% of isopropanol to solvent A greatly improved peak shape for analysis of wine samples (Fig. B). Please note
that histamine retention increased when isopropanol was in solvent A, since DI water is the strongest solvent and substituting it for isopropanol
reduces eluting power of the mobile phase.
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