Simultaneous determination of decitabine and vorinostat (Suberoylanalide hydroxamic acid, SAHA) by liquid chromatography tandem mass spectrometry for clinical studies.
A reverse-phase high-performance liquid chromatography method with electrospray ionization and detection by tandem mass spectrometry is described for the simultaneous quantitative determination of decitabine (5-aza-2'-deoxycytidine) and vorinostat (Suberoylanalide hydroxamic acid, SAHA) in human plasma. The method involves a simple acetonitrile precipitation step and centrifugation followed by injection of the supernatant onto a C18 150mmx2.1mm I.D., 3microm HPLC column at 36 degrees C. Separation of decitabine, SAHA and their respective internal standards was achieved with a gradient elution and detection was via the mass spectrometer operated in selected reaction monitoring mode. The method was within the defined validation parameters for linearity, repeatability, reproducibility and stability. The limit of detection was determined as 1.0 and 0.125ngml(-1) and lower limits of quantitation were 10 and 1ngml(-1) for decitabine and SAHA, respectively. Effects of sample preparation on stability were also evaluated in human plasma. For clinical sample handling tetrahydrouridine, an inhibitor of cytidine deaminase was found to help prevent decitabine degradation. The method is currently being used in clinical pharmacokinetic studies for the evaluation of decitabine and SAHA combination therapies.
Patel
Katan
K
Cancer Research UK. kpatel@staffmail.ed.ac.uk
Guichard
Sylvie M
SM
Jodrell
Duncan I
DI
D000970:Antineoplastic Agents / Q000032:analysis* / Q000097:blood; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000032:analysis; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D000077209:Decitabine; D005615:Freezing; D006801:Humans; D006877:Hydroxamic Acids / Q000032:analysis* / Q000097:blood; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions; D053719:Tandem Mass Spectrometry; D000077337:Vorinostat
D000970:Antineoplastic Agents; D006877:Hydroxamic Acids; D012996:Solutions; D000077337:Vorinostat; D000077209:Decitabine; D001374:Azacitidine