High-throughput quantitation of nefazodone and its metabolites in human plasma by high flow direct-injection LC-MS/MS.
A rapid, selective and sensitive high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS) method coupled with high flow direct-injection on-line extraction has been developed and validated for the simultaneous quantitation of nefazodone and its three active metabolites, hydroxynefazodone, triazole-dione (BMS-180492) and m-chlorophyenylpiperazine (mCPP) in human plasma. The method utilized d7-nefazodone, d7-hydroxynefazodone, d4-BMS-180492 and d4-mCPP as internal standards (IS). The plasma samples were injected into the LC-MS/MS system after simply adding the internal standard solution and centrifuging. The required extraction and chromatographic separation of the analytes were achieved on an Oasis HLB column (on-line extraction column, 1 mm x 50 mm, 30 microm) and a conventional Luna C8 column (analytical column, 4.6 mm x 50 mm, 5 microm). Detection was by positive ion electrospray tandem mass spectrometry. The total analysis run time for each sample was 2 min, which included the time needed for on-line extraction, chromatographic separation and LC-MS/MS analysis. The assay was validated for each analyte and the concentrations ranged from 2.0 to 500 ng/ml for nefazodone, hydroxynefazodone and mCPP and from 4.0 to 1000 ng/ml for BMS-180492, respectively. The assay was used for the high-throughput sample analysis of thousands of pharmacokinetic study samples and was proven to be rapid, accurate, precise, sensitive, specific and rugged.
Mao
Yan
Y
Bioanalytical and Discovery Analytical Sciences, Pharmaceutical Candidate Optimization, Bristol-Myers Squibb Pharmaceutical Research Institute, P.O. Box 191, New Brunswick, NJ 08903-0191, USA.
Huang
Mike-Qingtao
MQ
Xia
Yuan-Qing
YQ
Jemal
Mohammed
M
D018687:Antidepressive Agents, Second-Generation / Q000032:analysis* / Q000097:blood* / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D015394:Molecular Structure; D010879:Piperazines / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors; D014230:Triazoles / Q000032:analysis* / Q000097:blood* / Q000737:chemistry / Q000378:metabolism / Q000493:pharmacokinetics
D018687:Antidepressive Agents, Second-Generation; D010879:Piperazines; D014230:Triazoles; C406672:triazole-dione; C092333:hydroxynefazodone; C051752:nefazodone; C015068:1-(3-chlorophenyl)piperazine