Quantitative analysis of ES-285, an investigational marine anticancer drug, in human, mouse, rat, and dog plasma using coupled liquid chromatography and tandem mass spectrometry.
A method was developed for the quantitative analysis of the novel anticancer agent ES-285 (spisulosine; free base) in human, mouse, rat, and dog plasma using high-performance liquid chromatography/electrospray ionization tandem mass spectrometry in order to support pre-clinical and clinical studies with the drug. Sample preparation was carried out by protein precipitation with acetonitrile, containing isotopically labeled (d(3)) ES-285 as internal standard. Aliquots of 10 micro l of the supernatant were injected directly on to an Inertsil ODS-3 column (50 x 2.0 mm i.d., 5 micro m). Elution was carried out using methanol-10 mM ammonium formate (pH 4) in water (80 : 20, v/v) pumped at a flow-rate of 0.2 ml min(-1) with a run time of 8 min. Multiple reaction monitoring chromatograms obtained on an API365 triple-quadrupole mass spectrometer were used for quantification. The lower limit of quantitation (LLOQ) was 10 ng ml(-1) in human, mouse, rat, and dog plasma and the linear dynamic range extended to 500 ng ml(-1). A full validation of the method was performed in human plasma, and partial validations were performed in mouse, rat and dog plasma. Accuracies and precisions were <20% at the LLOQ concentration and <15% for all other concentrations in all matrices. ES-285 was stable during all steps of the assay. Thus far this method has been used successfully to analyze over 500 samples in pre-clinical trials, and will be implemented in the planned clinical phase I studies.
Stokvis
E
E
Department of Pharmacy and Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066 EC Amsterdam, The Netherlands.
Nan-Offeringa
L
L
Rosing
H
H
López-Lázaro
L
L
Aceña
J L
JL
Miranda
E
E
Lyubimov
A
A
Levine
B S
BS
D'Aleo
C
C
Schellens
J H M
JH
Beijnen
J H
JH
D000473:Alkanes / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D004355:Drug Stability; D015507:Drugs, Investigational / Q000032:analysis*; D006801:Humans; D007553:Isotope Labeling; D008055:Lipids / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D011786:Quality Control; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000473:Alkanes; D000970:Antineoplastic Agents; D015507:Drugs, Investigational; D008055:Lipids; C408363:spisulosine