The use of smaller particle-size columns can increase resolution and peak height, while decreasing run time.
This document presents an application note by Robert Ricker regarding chromatographic separation techniques. It highlights how transferring separations to columns with smaller particle sizes (specifically 3.5 µm) can result in increased resolution and peak height compared to larger particle sizes (5 µm).
| Column Type | ZORBAX 300SB-C8 Columns |
|---|---|
| Mobile Phase A | 95% ACN : 5% H2O, 0.1% TFA |
| Mobile Phase B | 5% ACN : 95% H2O, 0.085% TFA |
| Injection Volume | 10pL |
| Flow Rate | 1 mL/min |
| Column Temperature | 35°C |
| Detection Method | UV (245 nm) |
The document lists three column sizes: 4.6 x 50 mm, 4.6 x 150 mm, and 4.6 x 250 mm, all with a particle size of either 3.5 um or 5 um.
The method uses Mobile Phase A as 95% ACN with 5% H2O and 0.1% TFA, and Mobile Phase B as 5% ACN with 95% H2O and 0.085% TFA.
The procedure requires an injection of 10 uL at a flow rate of 1 mL/min with the column maintained at 35°C.