Simple microHPLC-UV method for the simultaneous determination of retinol and á-tocopherol in human plasma. Application to intrahepatic cholestasis of pregnancy

MARTINEFSKI M.,; SCIOSCIA S.,; CONTIN M.; SAMASSA P; LUCANGIOLI S.,; TRIPODI V.

ANALYTICAL METHODS

Royal sociaty of Chemistry

Año: 2014 vol. 6 p. 3365 - 3369

1759-9660

Abstract: a) Objectives: The development of a simple, rapid, sensitive, and accurate HPLC-UV method for the simultaneous determination of retinol and á-tocopherol in human plasma, suitable for the evaluation of these vitamins in pregnant women suffering intrahepatic cholestasis of pregnancy (ICP). b) Design and Methods: Chromatographic separation of vitamins was achieved with a microcolumn BDS HYPERSIL C18 (100 mm x 2.1mm, 2.4 ìm) using a C18 guard column. The developed and validated HPLC method proposed in this study was applied in the assessment of plasmatic concentrations of retinol and á-tocopherol in normal pregnancy (n=37) and in pregnancy complicated with ICP (n=19). c) Results: The validation of the method showed an excellent linearity for retinol and á-tocopherol (r2: 0.998 and 0.997), high sensitivity (LOD: 0.02 and 0.11ìM, LOQ: 0.07 and 0.36 ìM), high recovery (96.0%-104.0% and 99.3%-102.0%), respectively. The method requires low sample volume and all procedure of the sample preparation and analytical determination take less than 20 min to be completed. The evaluation of the results in ICP patients showed that á-tocopherol is significantly diminished compared to normal pregnant women (13.1±2.9 vs 22.4±2.2 µM, p