Make: Agilent Model: 1260 Infinity II
Detector: UV
Principle:
HPLC works on the principle of adsorption and partition chromatography.
Separation occurs due to the different degrees of interaction (polarity, ionic strength, hydrophobicity) between the stationary phase and the sample molecules.
Instrumentation
1. Solvent Reservoirs – Store mobile phases (aqueous/organic solvents).
2. Pump – Delivers the mobile phase at high pressure (up to 6000 psi).
3. Injector – Introduces a precise volume of sample.
4. Column – Packed with stationary phase (commonly silica-based C18, C8, etc.).
5. Detector – Detects separated compounds (commonly UV-Vis detector, PDA, Fluorescence).
6. Data System – Computer/software records chromatogram (peak vs retention time).
Uses of HPLC
Pharmaceuticals – Drug assay, impurity profiling, dissolution testing, stability studies.
Food & Beverages – Preservatives, sweeteners, vitamins, caffeine analysis.
Environmental – Water pollutant analysis, pesticide residues.
Forensics – Drug abuse testing, toxicology.
Biotechnology – Protein, peptide, amino acid analysis.
Tags:
Pharmacy