Determination of (4‐(1,3‐dioxo‐1,3‐dihydro‐2H‐isoindol‐ 2‐yl)‐N ′‐[(4‐ethoxyphenyl) methylidene] benzohydrazide, a novel antiinflammatory agent, in biological fluids by UPLC‐MS/MS: Assay development, validation and in vitro metabolic stability study
Ezzeldin, Muzaffar Iqbal, Mashooq A. Bhat, Mohammad Raish, Essam . 2017
A thalidomide analog, (4‐(1,3‐dioxo‐1,3‐dihydro‐2H‐isoindol‐2‐yl)‐N′‐[(4‐ethoxyphenyl)
methylidene] benzohydrazide), has been identified as a promising broad‐spectrum anti‐inflammatory
agent in previous study. In this study, a sensitive and selective UPLC‐MS/MS assay
was developed and validated for its determination in rat plasma samples. The chromatographic
separation was performed on an Aquity BEH C18 column using mobile phase comprising
of acetonitrile and 10mM ammonium acetate in the ratio of 85: 15, at flow rate of 0.3 mL/min.
The detection and quantification were performed in positive multiple reaction monitoring mode
by parent to daughter ion transition of 414.06 ˃ 148.05 for analyte and 411.18 ˃ 191.07 for
internal standard (risperidone), respectively using electrospray ionization source. The sample
extraction process consisted of liquid–liquid extraction method using diethyl ether as the
extracting solvent. The assay was validated by following FDA guidelines and all parameters were
found to be within acceptable limits. The linearity was between 10.1 and 2500 ng/mL and the
lower limit of quantification was 10.1 ng/mL. The reported results indicate that the assay could
meet the requirement for analysis of this compound in amounts expected to the present in actual
samples. Further, in vitro metabolic stability study was performed in rat liver microsomes by using
the validated assay.
A thalidomide analog, (4‐(1,3‐dioxo‐1,3‐dihydro‐2H‐isoindol‐2‐yl)‐N′‐[(4‐ethoxyphenyl)
methylidene] benzohydrazide), has been identified as a promising broad‐spectrum anti‐inflammatory…
A new series of 3,4,5-trimethoxyphenyl bearing
pyrazole (4a–g) and pyrazolo[3,4-d]pyridazine (5a–g)
scaffolds were synthesized in good yield. The newly synthesized
…