Introduction
An analytical HPLC system using microparticulate packing materials requires a lower system volume, higher operating pressure and high speed response. Rapid separation HPLC (RHPLC) is likely to become the new standard for high performance analysis, covering measurement in a wide range of applications from conventional HPLC to some types of UHPLC. The following is an analysis report for the measurement of fatty acids using a Unifinepak C18 column with the RHPLC system including a refractive index detector (RI-4035) for a semi-micro scale measurement.
Experimental
| Equipment | Conditions | ||
|---|---|---|---|
| Pump: | PU-4185 | Column: | A) Unifinepak C18 (4.6 mmI.D. x 150 mmL, 5 μm) |
| Pump option: | Degasser unit | B) Unifinepak C18 (2.0 mmI.D. x 150 mmL, 1.9 μm) | |
| Autosampler: | AS-4150 | C) Unifinepak C18 (2.0 mmI.D. x 30 mmL, 1.9 μm) | |
| Column oven: | CO-4060 | Eluent : | 0.1%TFA/Acetonitrile (20/80) |
| Detector: | RI-4035 | Flow rate: | A; 1.2 mL/min, B-C; 0.6 mL/min |
| Column temp.: | 40ºC | ||
| Injection volume: | A; 10 μL, B-C; 2 μL | ||
| Standard Sample: | Fatty acids mixture (1 mg/mL per component) |
Results
Fig. 1 Chromatograms of fatty acids were measured using the LC-4000 series RHPLC system with conventional HPLC and sub 2 μm UHPLC columns. The RI-4035 refractive index detector is optimized for lower flow rates and allows the acquisition of chromatograms with improved peak shape and a higher resolution separation. Unifinepak offers a common packing material in a range of particle sizes, which is independent of the measurement method, such as column size.
