Abstract
The release of free fatty acids from rat platelets, triggered by thrombin stimulation, was monitored by high-performance liquid chromatography (HPLC) after precolumn derivatization with monodansylcadaverine (MDC). A rapid filtration procedure was devised for the precise determination of free fatty acids released from aggregated platelets, instead of the conventional method using a stop solution or enzyme reactions. The fatty acids thus collected were derivatized with MDC in the presence of diethyl phosphorocyanidate (DEPC). The simultaneous separation of MDC derivatives of fatty acids was achieved on a reversed-phase TSKgel ODS-80 TM column within 60 min by linear gradient elution, using 0.2 M Tris-HCl buffer (pH 7.8)-methanol (50:50, v/v) and acetonitrile. The MDC derivatives were detected with excitation and emission wavelengths of 340 and 518 nm, respectively. The amounts of liberated fatty acids were in the range from 45.0 pmol for myristoleic acid (C 14:1) to 395.0 pmol for palmitic acid (C 16:0) per 1.9 · 10 7 platelets.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
