Development of a LC-MS method for quantification of FK-3000 and its application to in vivo pharmacokinetic study in drug development.
FK-3000 can inhibit proliferation of carcinomas and arrest the growth of carcinoma cells through cytotoxic (apoptosis induction) and cytostatic (cell cycle arrest) effects. A rapid and sensitive assay was developed and validated using liquid chromatography-mass spectrometry (LC-MS) for FK-3000 in rat plasma. FK-3000 was extracted with ethyl acetate from rat plasma samples, and the residue containing the FK-3000 was dried in a gentle stream of nitrogen and reconstituted with acetonitrile. The FK-3000 was quantified using high-performance liquid chromatography (HPLC; Waters Alliance 2695) with a reversed phase Gemini column (3 mm × 150 mm, 5 μm; Phenomenex, USA) and a Waters Micromass ZQ detector. FK-3000 and phenazine, an internal standard (IS), were analyzed by selected ion monitoring (SIM) at m/z transitions of 418.45 and 256, respectively. A lower limit of quantification (LLOQ) of 10 ng/mL was observed, with a linear dynamic range from 10 to 10,000 ng/mL (R>0.999). The accuracy, precision, recovery, matrix effects, and stability of the assay were deemed acceptable according to the FDA guidance for industry (bioanalytical method validation). The FK-3000 concentration was measured in plasma samples up to 6 h following FK-3000 administration at an oral dose of 20 mg/kg. The findings indicate that the assay method is suitable for routine pharmacokinetic (PK) studies of FK-3000 in rats.
Korea Institute of Toxicology, Yuseong, Daejeon 305-343, Republic of Korea.
Jong-Hwa
JH
Lee
Yoon-Jee
YJ
Chae
Kyeong-Ryoon
KR
Lee
Sung-Hoon
SH
Ahn
Joung-Wook
JW
Seo
Qing-Ri
QR
Jin
Young-Ah
YA
Woo
Gye-Won
GW
Lee
Soon-Chang
SC
Cho
Sung-Won
SW
Kwon
Dae-Hun
DH
Park
D000085:Acetates / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D000284:Administration, Oral; D000470:Alkaloids / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000592:standards; D056148:Chromatography, Reverse-Phase / Q000592:standards; D004355:Drug Stability; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000592:standards
D000085:Acetates; D000097:Acetonitriles; D000470:Alkaloids; D000972:Antineoplastic Agents, Phytogenic; C442856:FK3000; C007650:ethyl acetate; C032159:acetonitrile