C. Operating Conditions, HPLC Method For Quantifying Group 3 and 4 Analytes1
| Operating Parameter | Parameter Value | |
|---|---|---|
| Column | Alltech Altima C18, 150 × 4.6 mm, 5 μm particles | |
| Mobile phase | Component A = 0.05 M Potassium Phosphate Monobasic, pH 3.2 | |
| Component B = Acetonitrile | ||
| Mobile phase gradient | Time (min) | Proportion Component A |
| 0.0 | 70 | |
| 8.0 | 20 | |
| 10.0 | 20 | |
| 14.0 | 70 | |
| 18.0 | 70 | |
| Mobile phase flow rate | 1.0 mL/min | |
| Detection | UV at 215 nm | |
| Sample Size | 60 μL | |
↵1 Group 3 analytes included AN, AD EAB and BAB per Table I. Group 4 analytes included MBOH, EHB and BHB per Table I.