A sensitive and robust HPLC assay with fluorescence detection for the quantification of pomalidomide in human plasma for pharmacokinetic analyses.
Pomalidomide is a second generation IMiD (immunomodulatory agent) that has recently been granted approval by the Food and Drug Administration for treatment of relapsed multiple myeloma after prior treatment with two antimyeloma agents, including lenalidomide and bortezomib. A simple and robust HPLC assay with fluorescence detection for pomalidomide over the range of 1-500ng/mL has been developed for application to pharmacokinetic studies in ongoing clinical trials in various other malignancies. A liquid-liquid extraction from human plasma alone or pre-stabilized with 0.1% HCl was performed, using propyl paraben as the internal standard. From plasma either pre-stabilized with 0.1% HCl or not, the assay was shown to be selective, sensitive, accurate, precise, and have minimal matrix effects (<20%). Pomalidomide was stable in plasma through 4 freeze-thaw cycles (<12% change), in plasma at room temperature for up to 2h for samples not pre-stabilized with 0.1% HCl and up to 8h in samples pre-stabilized with 0.1% HCl, 24h post-preparation at 4°C (<2% change), and showed excellent extraction recovery (∼90%). This is the first reported description of the freeze/thaw and plasma stability of pomalidomide in plasma either pre-stabilized with 0.1% HCl or not. The information presented in this manuscript is important when performing pharmacokinetic analyses. The method was used to analyze clinical pharmacokinetics samples obtained after a 5mg oral dose of pomalidomide. This relatively simple HPLC-FL assay allows a broader range of laboratories to measure pomalidomide for application to clinical pharmacokinetics.
Clinical Pharmacology Program, National Cancer Institute, Bethesda, MD, USA.
Shandiz
S
Shahbazi
Clinical Pharmacology Program, National Cancer Institute, Bethesda, MD, USA.
Cody J
CJ
Peer
HIV/AIDS Malignancy Branch, National Cancer Institute, Bethesda, MD, USA.
Mark N
MN
Polizzotto
HIV/AIDS Malignancy Branch, National Cancer Institute, Bethesda, MD, USA.
Thomas S
TS
Uldrick
Clinical Pharmacology Program, National Cancer Institute, Bethesda, MD, USA.
Jeffrey
J
Roth
HIV/AIDS Malignancy Branch, National Cancer Institute, Bethesda, MD, USA.
Kathleen M
KM
Wyvill
HIV/AIDS Malignancy Branch, National Cancer Institute, Bethesda, MD, USA.
Karen
K
Aleman
Celgene Corporation and Celgene Global Health, Summit, NJ, USA.
Jerome B
JB
Zeldis
HIV/AIDS Malignancy Branch, National Cancer Institute, Bethesda, MD, USA.
Robert
R
Yarchoan
Clinical Pharmacology Program, National Cancer Institute, Bethesda, MD, USA. Electronic address: figgw@helix.nih.gov.
William D
WD
Figg
D001681:Biological Assay / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005453:Fluorescence; D006801:Humans; D059625:Liquid-Liquid Extraction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013792:Thalidomide / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics
D013792:Thalidomide; C467566:pomalidomide
Fluorescence; HPLC; Liquid extraction; Pharmacokinetics; Pomalidomide