High Performance Liquid Chromatography - Pace ….pptVIP

High Performance Liquid Chromatography Chem. 331 Introduction HPLC is a form of liquid chromatography used to separate compounds that are dissolved in solution. HPLC instruments consist of a reservoir of mobile phase, a pump, an injector, a separation column, and a detector. Compounds are separated by injecting a sample mixture onto the column. The different component in the mixture pass through the column at differentiates due to differences in their partition behavior between the mobile phase and the stationary phase. The mobile phase must be degassed to eliminate the formation of air bubbles. HPLC system FOUR TYPES OF LIQUID CHROMATOGRAPHY Partition chromatography Adsorption, or liquid-solid chromatography Ion exchange chromatography Size exclusion, or gel, chromatography COMPOSITION OF A LIQUID CHROMATOGRAPH SYSTEM Solvent Solvent Delivery System (Pump) Injector Sample Column Detectors (Diode Array) Waste Collector Recorder (Data Collection) Picture of HPLC instrument Uses of HPLC This technique is used for chemistry and biochemistry research analyzing complex mixtures, purifying chemical compounds, developing processes for synthesizing chemical compounds, isolating natural products, or predicting physical properties. It is also used in quality control to ensure the purity of raw materials, to control and improve process yields, to quantify assays of final products, or to evaluate product stability and monitor degradation. In addition, it is used for analyzing air and water pollutants, for monitoring materials that may jeopardize occupational safety or health, and for monitoring pesticide levels in the environment. Federal and state regulatory agencies use HPLC to survey food and drug products, for identifying confiscated narcotics or to check for adherence to label claims. HPLC Chromatograph injectors The function of the injector is to place the sample into the high-pressure flow in as narrow volume as possible so that the sample