TY - JOUR
T1 - Rapid and simultaneous determination of efavirenz, 8-hydroxyefavirenz, and 8,14-dihydroxyefavirenz using LC-MS-MS in human plasma and application to pharmacokinetics in healthy volunteers
AU - Kim, Kwon Bok
AU - Kim, Hyunmi
AU - Jiang, Fen
AU - Yeo, Chang Woo
AU - Bae, Soo Kyung
AU - Desta, Zeruesenay
AU - Shin, Jae Gook
AU - Liu, Kwang Hyeon
PY - 2011/2
Y1 - 2011/2
N2 - We developed a rapid and sensitive method for determining efavirenz, 8-hydroxyefavirenz, and 8,14-dihydroxyefavirenz in human plasma simultaneously using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Three compounds and ritonavir, an internal standard, were extracted from plasma using ethyl acetate in the presence of 0.1 M sodium carbonate after incubation of β-glucuronidase (500 U). After drying the organic layer, the residue was reconstituted in mobile phase (acetonitrile:20 mM ammonium acetate, 90:10, v/v) and injected onto a reversed-phase C18 column. The isocratic mobile phase was eluted at 0.2 mL min-1. The ion transitions monitored in multiple reaction-monitoring mode were m/z 314 → 244, 330 → 258, 346 → 262, and 721 → 296 for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The retention time is 1.93, 1.70, 1.52, and 1.82 min for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The coefficients of variation of the assay precision were less than 10.7%, and the accuracy was 90-111%. The lower limits of quantification (LLOQ) were 5 ng mL-1 for efavirenz and 8-hydroxyefavirenz. This method was used to measure the plasma concentrations of efavirenz and its metabolites from healthy volunteers after a single 600 mg oral dose of efavirenz. This analytical method is a very rapid, sensitive, and accurate to determine the pharmacokinetic profiles of efavirenz including its metabolites.
AB - We developed a rapid and sensitive method for determining efavirenz, 8-hydroxyefavirenz, and 8,14-dihydroxyefavirenz in human plasma simultaneously using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Three compounds and ritonavir, an internal standard, were extracted from plasma using ethyl acetate in the presence of 0.1 M sodium carbonate after incubation of β-glucuronidase (500 U). After drying the organic layer, the residue was reconstituted in mobile phase (acetonitrile:20 mM ammonium acetate, 90:10, v/v) and injected onto a reversed-phase C18 column. The isocratic mobile phase was eluted at 0.2 mL min-1. The ion transitions monitored in multiple reaction-monitoring mode were m/z 314 → 244, 330 → 258, 346 → 262, and 721 → 296 for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The retention time is 1.93, 1.70, 1.52, and 1.82 min for efavirenz, 8-hydroxyefavirenz, 8,14-dihydroxyefavirenz, and ritonavir, respectively. The coefficients of variation of the assay precision were less than 10.7%, and the accuracy was 90-111%. The lower limits of quantification (LLOQ) were 5 ng mL-1 for efavirenz and 8-hydroxyefavirenz. This method was used to measure the plasma concentrations of efavirenz and its metabolites from healthy volunteers after a single 600 mg oral dose of efavirenz. This analytical method is a very rapid, sensitive, and accurate to determine the pharmacokinetic profiles of efavirenz including its metabolites.
KW - 8,14-Dihydroxyefavirenz
KW - 8-Hydroxyefavirenz
KW - Column liquid chromatography-tandem mass spectrometry
KW - Efavirenz
KW - Human plasma
UR - http://www.scopus.com/inward/record.url?scp=79951770887&partnerID=8YFLogxK
U2 - 10.1007/s10337-010-1882-5
DO - 10.1007/s10337-010-1882-5
M3 - Article
AN - SCOPUS:79951770887
SN - 0009-5893
VL - 73
SP - 263
EP - 271
JO - Chromatographia
JF - Chromatographia
IS - 3-4
ER -