Automated high-performance liquid chromatographic determination of plasma free fatty acids using on-line derivatization with 9-bromomethylacridine based on micellar phase-transfer catalysis.
The on-line use of micellar phase-transfer catalysis is described for the automated reversed-phase high-performance liquid chromatographic (RP-HPLC) determination of free fatty acids in plasma; minimum manual sample handling is involved. After diluting plasma ten-fold with the aqueous micellar system, which contains 25 mM of the non-ionic surfactant, Arkopal N-130 and 6 mM of the ion-pair agent tetrakis(decyl)ammonium bromide, the reaction of the fatty acids with the fluorophore 9-bromomethylacridine is complete within 5 min at 60 degrees C. Prior to RP-HPLC separation, interfering proteins are removed using an on-line filter and a column-switching unit. More than 100 samples can be injected onto a single pre-column. The detection limit is ca. 300 nM; the precision is better than 3% using an internal standard.
van der Horst
F A
FA
Department of Pharmaceutical Analysis, University of Utrecht, The Netherlands.
Post
M H
MH
Holthuis
J J
JJ
Brinkman
U A
UA
D000166:Acridines; D002623:Chemistry Techniques, Analytical; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D005230:Fatty Acids, Nonesterified / Q000097:blood*; D005453:Fluorescence
D000166:Acridines; D005230:Fatty Acids, Nonesterified; C063815:9-bromomethylacridine