Table 1.
Derivatization of standards and samples.
| Standards (from 76 to 608 nM) | MDA-Hb | |
|---|---|---|
| Sample volume | 100 μL TMP | 100 μL |
| Derivatization | 100 μL DAN 5.8 mM in HCl 2.4 N + internal standard | |
|
| ||
| Mixing, incubation for 180 min at 37°C | ||
|
| ||
| Hb precipitation | 100 μL MPA 10%, mixing, centrifugation 3 min, 15000 × g | |
| 200 μL supernatant for the next step | ||
| Stabilization at pH 2.0* | 100 μL (NaOH 1 N + KH2PO4 300 mM pH 2.0) | |
*At every new preparation of both reagents, the volumes are adjusted to obtain pH 2.0 with a fixed volume of 300 μL (DAN: diaminonaphthalene; MDA: malondialdehyde; MPA: metaphosphoric acid; TMP: tetramethoxypropane).