Validation of an analytical method for a potent antitumor agent, TZT-1027, in plasma using liquid chromatography-mass spectrometry.
A sensitive and specific analytical method for a potent antitumor agent, TZT-1027, in plasma has been developed using liquid chromatography-mass spectrometry (LC-MS) with [2H4]TZT-1027 as an internal standard (I.S.). A plasma sample was purified by solid-phase extraction on a C18 cartridge, followed by solvent extraction with diethyl ether. The extract was then injected into the LC-MS system. Chromatography was carried out on a C18 reversed-phase column using acetonitrile-0.05% trifluoroacetic acid (TFA) (55:45) as a mobile phase. Mass spectrometric analysis was performed in atmospheric pressure chemical ionization (APCI) mode with positive ion detection, and the protonated molecular ions ([M+H]+) of TZT-1027 and I.S. were monitored to allow quantitation. The method was applied to the determination of TZT-1027 in human, monkey, dog, rat and mouse plasma. As far as the sample preparation was concerned, good recoveries (73.5-99.1%) were obtained. The calibration curves were linear over the range of 0.25-100 ng per 1 ml of human, dog and rat plasma, per 0.5 ml of monkey plasma, and per 0.1 ml of mouse plasma. From the intra- and inter-day accuracy and precision, the present method satisfies the accepted criteria for bioanalytical method validation. TZT-1027 was stable when stored below -15 degrees C for 6 months in human plasma and for 3 weeks in plasma from other species. TZT-1027 was also stable in plasma through at least three freeze-thaw cycles.
Ochiai
H
H
Pharmacokinetics Research Department, Teikoku Hormone Mfg. Co. Ltd., Kawasaki, Kanagawa, Japan.
Mori
H
H
Murata
H
H
Seki
T
T
Araki
K
K
Kawabe
Y
Y
Miyazaki
K
K
Tsukamoto
K
K
Iwamura
S
S
D000818:Animals; D000970:Antineoplastic Agents / Q000097:blood*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009842:Oligopeptides / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000970:Antineoplastic Agents; D009842:Oligopeptides; C105791:soblidotin