Basics of HPLC:
It is now the most versatile and widely used technique by chemists for the separation, qualitative identification and quantitative determination of species in a variety of organic, inorganic, biological and other complex materials. It is a type of elution chromatography while the sample, a mixture of solutes, is within a liquid solvent and mobile phase. The method is also known through other synonyms like as high resolution chromatography, speed chromatography and high efficiency chromatography and is considered as the most sensitive method along with continuous main developments. HPLC is able to separate macromolecules and ionic species, polymeric materials, labile natural products, and a huge variety of other high molecular weight polyfunctional groups. HPLC separations are based on specific interactions between sample molecules with both the stationary and mobile phases. A large variety of stationary phases available in HPLC allow a great variety of selective interactions causing better separations.
Objectives
After studying this Unit, you should be able to
- Elaborates the meaning of high performance liquid chromatography,
- Distinguish between classical liquid chromatography and HPLC,
- Explain the basic principle and working of HPLC,
- Define various components of instrumentation including stationary and mobile phases,
- Define characteristics of stationary phases in various modes including bonded phase,
- Understand the solvent delivery system, features of mobile phase and elution gradient,
- Know various detectors used in HPLC,
- Know about versatility and advantages of HPLC,
- Understand about various interfaces while using mass spectrometer as detector, and
- Know about applications of HPLC for the analysis of a variety of solutes.