SOLID-PHASE EXTRACTION COMBINED WITH UHPLC-MS/MS METHOD FOR DETERMINATION OF REMIFENTANIL IN HUMAN WHOLE BLOOD

A sensitive and reliable method have been developed and validated using solid phase extraction (SPE) combined with ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to determine remifentanil in human blood. Quantification was performed by external standard calibration (r = 0.9991, n = 5). Remifentanil was separated on an ACQUITY UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 mu m) and analyzed in positive-ion electrospray-ionization (ESI+) mode. The mobile phase was methanol and water with a gradient elution program. The total run time was 4.5 min and injection volume was 5 mu L. Limit of detection (LOD) and limit of quantitation (LOQ) were 0.20 ng/mL and 0.60 ng/mL, respectively. Remifentanil was eluted at 1.89 min. Recoveries of remifentanil ranged between 91.88%-93.79%. The intra-day and the inter-day precision (RSD) for remifentanil were 2.51%-3.48% and 2.76%-3.78%, respectively. The performance of the method was successfully verified for the determination of remifentanil in human blood.