HPLC‐APCI‐MS Determination of Free Fatty Acids in Tibet Folk Medicine Lomatogonium rotatum with Fluorescence Detection and Mass Spectrometric Identification

Abstract

A simple and sensitive method for the determination of free fatty acids (FFAs) using acridone‐9‐ethyl‐p‐toluenesulfonate (AETS) as a fluorescence derivatization reagent by high performance liquid chromatography (HPLC) has been developed. Free fatty acid derivatives were separated on an Eclipse XDB‐C₈ column with a good baseline resolution and detected with the fluorescence of which excitation and emission wavelengths of derivatives were set at λ_ex=404 and λ_em=440 nm, respectively. Identification of 19 fatty acid derivatives was carried out by online post‐column mass spectrometry with an atmospheric pressure chemical ionization (APCI) source under positive‐ion detection mode. Nineteen FFAs from the extract of Lomatogonium rotatum are sensitively determined. The results indicate that the plant Lomatogonium rotatum is enriched with an abundance of FFAs and FFAs of higher contents, which mainly focus on even carbon atoms, C₁₄, C₁₆, and C₁₈. The validation of the method including linearity, repeatability, and detection limits was examined. Most linear correlation coefficients for fatty acid derivatives are >0.9989, and detection limits (at signal‐to‐noise of 3:1) are 12.3–43.7 fmol. The relative standard deviations (RSDs) of the peak areas and retention times for 19 FFAs standards are <2.24% and 0.45%, respectively. The established method is rapid and reproducible for the separation determination of FFAs from the extract of Lomatogonium rotatum with satisfactory results.

Keywords:

• HPLC‐APCI‐MS
• FFAs
• AETS
• Fluorescence detection
• Derivatization
• Lomatogonium rotatum

Acknowledgments

Support for this research was provided by the Chinese Academy of Sciences (the Talent Cultivation Plan “Hope of the West China” and the Knowledge Innovation Program).