abstractAn analytical methodology based on cloud point extraction (CPE) coupled to Ultra-Fast Liquid Chromatography and electrospray tandem mass spectrometry (UFLC-MS/MS) was developed for analysis ofAbacavir (ABC), Efavirenz (EFV), Lamivudine (3 TC) and Nelfinavir (NFV) in human plasma. It is the firsttime that CPE was used for extraction of antiretrovirals (ARV) from plasma. The effects of relevant physicchemical variables on analytical response of each ARV, including pH, surfactant concentration, equilibration time and temperature, were study and optimized; as well as its coupling to UFLC-ESI-MS/MS.Under optimized conditions, the resulting methodology was as follows: a 500 mL aliquot of humanplasma was diluted with 2 mL deionized water in a 10 mL centrifuge tube. A 500 mL aliquot Triton X-1145% w/v was added and homogenized using a vortex stirrer. The resulting cloudy solution was kept at65 C for 20 min for promoting the condensation of surfactant micelles. Then it was centrifuged at3000 g for 5 min for separation of the surfactant-rich phase. After discarding the aqueous supernatant,400 mL ACN were added to the remaining surfactant rich phase and centrifuged in order to precipitateproteins and separate them. A 150 mL aliquot of the supernatant was transferred to 2 mL vial and furtherdiluted with 400 mL deionized water. A 30 mL aliquot of the so-prepared solution was injected andanalyzed into the UFLC-MS/MS. The method detection limits for ABC, EFV, 3 TC and NFV under optimizedconditions were 31, 77, 57 and 21 ng mL1, respectively. The RSD% for the studied analytes were <15%,except at the LOQ, which were <19%. Recovery values ranged from 81 to 107%. The proposed methodology was successfully applied for the analysis of ABC, EFV, 3 TC and NFV in human plasma within theconcentration range of 43e6816, 125e4992, 81e3248 and 49e7904 ng mL1, respectively. Under optimized working conditions the proposed analytical methodology meets standard requirements of international guidelines, which makes it suitable for pharmacokinetic studies of the four ARV, as well as fortherapeutic monitoring of HIV patients
การแปล กรุณารอสักครู่..