Parent drug or active metabolite?Should the assay focus on the parent compound only, or are active metabolites, prodrug forms, or endogenous-hormone background levels also relevant?
Hormone-axis context?Does the study involve HPA-axis, thyroid-axis, or gonadal-axis feedback loops that require multiple-analyte or time-course profiling?
Peptide or protein stability?For insulin, growth factors, or hematologic biologics, are peptide-level stability, aggregation, or matrix-specific recovery considerations needed?
Steroid metabolite profiling?Is steroid-metabolism context important for interpreting parent-drug exposure and endogenous-hormone suppression?
Glucose or lipid markers?Are glucose metabolism, insulin sensitivity, or lipid pathway biomarkers needed alongside drug concentration data?
Matrix-specific quantification?Do hormone assays or growth factor measurements require method development tailored to plasma, serum, or other specific matrices?