This Application Note describes the reverse-phase HPLC method for the analysis of fatty acids using Agilent HPLC systems.
This document details a method for the analysis of lipids, specifically fatty acids, which are important components in the food industry. The text explains that identifying and quantifying fatty acids is complicated by the lack of a strong UV chromophore, necessitating specific detection methods.
The note proposes a solution using a PLRP-S column coupled with an Agilent Evaporative Light Scattering Detector (ELSD). This combination allows for the separation of fatty acids across a wide range of pH levels and eluents, providing a stable baseline for detection. Specific experimental conditions, including column dimensions, eluent composition, and gradient settings, are provided to achieve good separation of various fatty acids.
| Eluent A | 60 mM Acetic acid |
|---|---|
| Eluent B | ACN |
| Eluent C | THF |
| Flow Rate | 0.5 mL/min |
| Detection Neb Temperature | 80 °C |
| Detection Evap Temperature | 70 °C |
The Agilent evaporative light scattering detector is an ideal detector for the analysis of fatty acids.
These columns are robust enough to be stable at pH 1-14.