Selection depends on analyte polarity, solubility, and required
sensitivity.
λ-max Determination
Wavelength where compound shows maximum absorbance.
Mobile Phase Optimization
Adjust solvent ratio to achieve proper separation.
Column Selection (C18 / C8)
Stationary phase selection based on compound nature.
Flow Rate &
Injection Volume
Optimized to improve resolution and peak shape.
Quantitative determination of active compound.
Ensures system performance before analysis.
Closeness to true value.
Repeatability and reproducibility.
Response proportional to concentration.
No interference from impurities.
Method stability under small changes.
Detection and quantification limits.
Stability of prepared solution over time.
Forced degradation under stress conditions.
Complete validation report with calculations, graphs, and chromatograms.
