HPLC
High-Performance Liquid Chromatography (HPLC) is a powerful analysis method used in chemistry, biochemistry and pharma. It uses high-pressure liquid flow to elision, label and quantitate elements in a sample through a column filled with stationary phase. HPLC is highly resolvable, sensitive and fast, and it's the key to the purification of dense mixtures. Learn more about pharmaceutical analysis.
Importance of HPLC in the Pharmaceutical Industry
For the pharmaceutical industry, HPLC is indispensable. It's used to investigate the structure of drugs and to confirm purity, but also to validate safety, efficacy and quality of medicines. HPLC allows the separation and quantification of active components and contaminants in drugs in detail which aids drug makers in quality control and stability studies. Further, HPLC is also an integral part of drug development where new drug synthesis, formulation optimization and quality assessment of final products are monitored by HPLC.
Fundamentals of HPLC
Basic Principles of HPLC
HPLC is a highly sophisticated process that separates, analyzes and purifies difficult mixtures. This is the basic logic of HPLC — the differential interactions of the stationary and mobile phases, to separate components in a sample. The sample in HPLC is carried through a chromatography column filled with the stationary phase, while the mobile phase moves the sample through the column. Material is pulled apart by hydrophobicity, polarity, etc. and it moves at various speeds to separate.
Key Components of an HPLC System
These are the primary parts of an HPLC system: solvent reservoirs, pumps, sample injectors, columns, detectors and processing units. All these pieces are necessary for samples to be separated and analysed.
Components of an HPLC System
Types of HPLC
Normal-Phase HPLC: Uses polar stationary and non-polar mobile phases. It's commonly used for the separation of non-polar and weakly polar elements.
Reversed-Phase HPLC: Uses non-polar stationary and polar mobile phases. This is a good way to dissociate polar molecules.
Ion Exchange Chromatography (IEC): Separates charged molecules by ion exchange. It's used extensively to study biomolecules.
Size Exclusion Chromatography (SEC): Separates molecules by molecular size, and can be used for protein, polysaccharide, and other macromolecules.
Each type of HPLC has its applications and pros and cons. For instance, normal phase HPLC is usually used for non-polar compound separation and reversed phase HPLC is widely used in pharma analysis and environmental monitoring.
Because of its high resolution, sensitivity and generalizability, HPLC is widely used in chemistry, biology, pharmaceuticals, and environmental studies.