Novel stationary phasis in HPLC for the analysis of biologically active compounds for clinical applications-determination of retinoic acid

All-trans retinoic acid (ATRA) and 13-cis-retinoic acid (13-cis RA) are naturally occurring retinoids. All-trans-retinoic acid plays multiple roles in reproduction, embryogenesis, cellular differention, vision and immune response and is also used therapeutically in acute promyelocytic leukemia. Tocopherols are major serum antioxidants. Both retinoids and tocopherols are studied as chemopreventive and therapeutic agents in cancer patients. In the present study, a simple and rapid HPLC method was developed for simultaneous determination of fat-soluble vitamins and retinoic acids (retinol, all-trans retinoic acid, 13-cis retinoic acid, gamma- and alpha-tocopherol) in human serum using monolithic technology. The monolithic column Chromolith Performance RP-18e (100 mm × 4.6 mm) was operated at ambient temperature. Mobile phase (MF) consisted of mixture Acetonitrile (ACN) : 1% ammonium acetate in water (AMC) at pH 7.0 starting with ACN : AMC 98:2 (column pre- treatment) at flow rate 2ml/min following 4,2 minutes ACN:AMC 95:5 at the flow rate 1.5 ml/min than 3 minutes flow rate 3,2ml/min with the same mobile phase. Detection and identification was performed using photodiode array detector. Retinol, 13-cis- and all-trans- retinoic acid were monitored at 325 nm. Alpha- and gamma-tocopherol were carried out..