Analysis of Fatty Acids using the LC-4000 Series Rapid Separation HPLC

Download PDF August 18, 2022


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.


Pump:PU-4185Column: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-4060Eluent :0.1%TFA/Acetonitrile (20/80)
Detector:RI-4035Flow 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)


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.

About the Author

Mr. Akao, is the Applications Group Manager at JASCO Corporation, with many years at the company working on applications in electronic and vibrational spectroscopy, and is an expert in IR and Raman spectroscopy.