Chromatographic protocols
The aqueous solvent (A) was 0.1% acetic acid in water (v/v) and the organic solvent (B) was 0.1% acetic acid in acetonitrile (v/v). The autosampler chamber was held at 6°C, and columns were held at 25°C for all experiments. The protocol for BPA in vivo is based on that of Yokota et al. (1999).
| Assay | Column | λ | Flow Rate | Elution Program |
|---|---|---|---|---|
| nm | ml/min | |||
| Naproxen in vitro | Zorbax Eclipse XDB-C18 (150 × 4.6 mm, 5µm) | 225 | 1.50 | 30% B from 0 to 5 minutes |
| Linear increase to 70% B from 5 to 5.5 minutes | ||||
| Linear increase to70% B from 5.5 to 10 minutes | ||||
| BPA in vitro | Zorbax Eclipse XDB-C18 (150 × 4.6 mm, 5µm) | 280 | 0.50 | 15% B from 0 to 4.5 minutes |
| Linear increase to 60% B from 4.5 to 5 minutes | ||||
| Linear increase to 60% B from 5 to 16 minutes | ||||
| BPA in vivo | Tosoh TSKgel ODS 80TM (250 × 4.6 mm, 5 µm) | 280 | 1.00 | Isocratic 35% B |