Workbench software

The cocktail probes and their corresponding CYP-specific metabolites will be quantified in plasma using high-performance liquid chromatography/tandem mass spectrometry method, which was previously fully validated and described in [15] (link). The protein precipitation by acetonitrile will be used for plasma extraction.
Clinical chemistry analysis, including creatinine, AST, ALT, FBS, and HbA1c, will be analyzed, primarily at the Department of Medical Biochemistry at IMAM KHOMEINI hospital complexes.
The plasma levels of proinflammatory cytokines including IL-1β, IL-6, and TNF-α will be quantified by electrochemiluminescence immunoassays using the V-PLEX Proinflammatory Panel 1 Human Kit, QuickPlex SQ120 Imager, and WORKBENCH software (MSD, Rockville, MD). The genetic polymorphism of CYP450 enzymes including CYP2B6, CYP2C9, CYP2C19 and CYP2D6 will be evaluated. Genomic DNA will be extracted from whole blood samples throughout the study in both groups. DNA samples will be used for genetic analyses associated with mentioned CYP450s by using semi-quantitative RT-PCR or other appropriate methods [14 (link),15] (link).

Blood samples for the quantification of proinflammatory cytokines were kept on ice and rapidly sent to the research laboratory to be centrifuged at 1100× g (10 min at 4 °C) within 1 hour of sampling. Plasma was then aliquoted and stored at −80 °C until use. Levels of inflammatory markers INF-γ, IL-1β, IL-6, and TNF-α were quantified by electrochemiluminescence immunoassays using the V-PLEX Proinflammatory Panel 1 Human Kit, QuickPlex SQ120 Imager, and WORKBENCH software (MSD^®, Rockville, MD).