Microdose clinical trial: quantitative determination of nicardipine and prediction of metabolites in human plasma.
A sample treatment procedure and high-sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for quantitative determination of nicardipine in human plasma were developed for a microdose clinical trial with nicardipine, a non-radioisotope labeled drug. The calibration curve was linear in the range of 1-500 pg/mL using 1 mL of plasma. Analytical method validation for the clinical dose, for which the calibration curve was linear in the range of 0.2-100 ng/mL using 20 microL of plasma, was also conducted. Each method was successfully applied to making determinations in plasma using LC/MS/MS after administration of a microdose (100 microg) and clinical dose (20 mg) to each of six healthy volunteers. We tested new approaches in the search for metabolites in plasma after microdosing. In vitro metabolites of nicardipine were characterized using linear ion trap-fourier transform ion cyclotron resonance mass spectrometry (LIT-FTICRMS) and the nine metabolites predicted to be in plasma were analyzed using LC/MS/MS. There is a strong possibility that analysis of metabolites by LC/MS/MS may advance to utilization in microdose clinical trials with non-radioisotope labeled drugs.
Yamane
Naoe
N
Nishiwaki Laboratory, JCL Bioassay Corporation, Hyogo, Japan. n.yamane@jclbio.com
Takami
Tomonori
T
Tozuka
Zenzaburo
Z
Sugiyama
Yuichi
Y
Yamazaki
Akira
A
Kumagai
Yuji
Y
D000959:Antihypertensive Agents / Q000097:blood / Q000378:metabolism*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical; D004355:Drug Stability; D006801:Humans; D008297:Male; D013058:Mass Spectrometry / Q000379:methods; D008862:Microsomes, Liver / Q000378:metabolism*; D009529:Nicardipine / Q000097:blood / Q000378:metabolism*; D004364:Pharmaceutical Preparations / Q000097:blood / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods
D000959:Antihypertensive Agents; D004364:Pharmaceutical Preparations; D009529:Nicardipine