Chromatography and Analytical Techniques

Partition chromatography is method of separation in which the components present in the mixture get distributed more likely into two liquid phases because of differences in partition coefficients. In Partition chromatography, the molecules are separated in between two phases i.e. both stationary phase and mobile phase are in same phase. So molecules get dispersed into either phase preferentially.  The partition chromatography is the basic principle involved in many separation techniques like high performance liquid chromatography and gas chromatography. Partition chromatography is basically understood as a method of separation of solutes utilizing the partition of the solutes between two liquid phases. Paper chromatography and High performance TLC also works on the principle of the same

Ion exchange chromatography (IEX) separates molecules present in the mixture based on differences in their net surface charge. Ion exchange chromatography playing an important role in the separation and purification of biomolecules. Today, IEX is one of the most frequently used separation techniques for purification of amino acids, proteins, peptides, and nucleic acids. Ion Exchange Chromatography depends up on charge-charge interactions between the proteins in the sample and the charges immobilized on the resin of choice. Loading samples in buffers of low ionic strength makes ion exchange chromatography an excellent purification step after hydrophobic interaction chromatography

Molecular exclusion chromatography also called or named as Size exclusion chromatography (SEC) separates molecules depending on their molecular size by filtration through a gel. The gel consists of spherical beads containing pores of a particular size distribution. Separation techniques occurs when molecules present in the mixture, of different sizes are included or removed from the pores within the matrix. Small molecules and large molecules are separated based on entering in to the pores. Consequently, molecules separate based on their size as they pass through the column and are eluted in order of decreasing molecular weight (MW)

Special techniques in analytical chemistry also play an important role in the analysis of many chemical compounds. Physical separation techniques are the most common, least expensive and safest. Component separation is also relatively easy but tends to be more expensive. The specialty chemical and pharmaceutical industries are the primary users of component separation techniques. Component separation is only now being applied to hazardous wastes and many processes are still in the demonstration stage. Chemical separation techniques are the most expensive and pose the greatest potential hazard. In Hydrophobic Interaction Chromatography (HIC) the proteins are separated according to differences in the amount of exposed hydrophobic amino acids

Spectroscopy relates to the scattering an object's light into its component colors (i.e. energies). By performing the spectroscopy of an object, astronomers can find out the physical properties of that object (like temperature, mass, luminosity and composition). Historically spectroscopy was defined as the use of visible light dispersed according to its wavelength. After that this concept was developed greatly to comprise any kind of measurement of a quantity as a function of either wavelength or frequency. The various devices used in Spectrometry are spectrometers, spectrophotometers, spectrographs or spectral analyzers. Absorption of infrared radiation brings about changes in molecular vibrations within molecules and measurements of the ways in which bonds vibrate gives rise to infrared spectroscopy. Mass spectrometry is other analytical technique, which separates the ions according to their mass/charge ratio

Affinity chromatography is one of the important separation techniques based on a specific binding interaction between an immobilized ligand and its binding partner. Affinity chromatography is a type of liquid chromatography that makes use of interactions for the identification and specific analysis of sample components. Examples include antibody and antigen, enzyme and substrate, and enzyme and inhibitor interactions. The degree of purification can be quite high in a purification strategy. This present track provides an knowledge of affinity chromatography of proteins. Immune affinity chromatography is one of the important types of affinity chromatography. Immune affinity chromatography technique is more commonly performed for the identification, quantification, or purification of antigens

Gas chromatography is a generally a word used to give information about the group of analytical separation techniques used to analyze volatile materials in the gas phase. In this chromatography, the components of a sample are dissolved in a solvent and vaporized which results in the separation of analytics by differentiating the sample between two phases. Gas chromatography is one of the important types of chromatography that does not use the mobile phase for reacting with the analytic. If the stationary phase consists of solid adsorbant, termed gas-solid chromatography or a liquid on an inert support, termed gas-liquid chromatography (GLC). Gas Chromatography Mass Spectrometry (GC-MS) is the advanced technique of Gas Chromatography. Gas chromatography can be used in many different fields such as pharmaceutical industry, cosmetology and even environmental toxins

High Performance Liquid Chromatography (HPLC) is different and another type of column chromatography that pumps a sample mixture or analyte in a solvent at high pressure through a column with chromatographic packing material. HPLC has the ability to analyse, and separate compounds that would be present in any sample that can be dissolved in a liquid in trace concentrations. Because of this advantage, HPLC is used in a variety of industrial and scientific applications, such as pharmaceutical industry, environmental, forensic science, and chemicals. High Performance Liquid Chromatography has brought lot of advantages in the department of food analysis and also in the analysis of various fat soluble vitamins. HPLC is also used in DNA fingerprinting and bioinformatics

There has been growing interest around the globe in various change separation techniques, in key areas of physical sorption, distillation, membranes, absorption and heat exchange. These special technologies in chromatography include Reversed-phase chromatography, Normal-phase chromatography, Two-dimensional chromatography and pyrolysis chromatography. In normal-phase chromatography, the stationary phase is exactly opposite to the mobile phase which is nonpolar. Reversed-phase chromatography (RPC) is another liquid chromatography method where the mobile phase is significantly more polar than the stationary phase. Chiral chromatography which definitely means the identification of stereoisomers. Pyrolysis gas chromatography mass spectrometry is another type of chemical analysis in which the sample is heated to decomposition to produce smaller molecules which are analyzed by gas chromatography and detected using mass spectrometry

Chromatography, different method for identifying and separating the components, or solutes, of a mixture on the basis of the relative amounts of each solute distributed between a moving fluid streams, called the mobile phase, and a contiguous stationary phase. Liquid-chromatography is one of the mostly employed and useful techniques in pharmaceutical industry for sample preparation. Another one is electrophoretic separation technique designed for rapid and selective sample analysis. Immune affinity chromatography (IAC) is a different method of Liquid Chromatography in which the stationary phase consists of an antibody or it might include antibody-related reagent. This technique involves a unique sub type of affinity chromatography, in which a biologically related binding agent is taken for the selective separation or analysis of a target compound. Capillary electrophoresis is other method which identifies and distributes ions depending on their electrophoretic mobility with the use of an applied voltage. Compact mass spectrometer is widely used in the food safety, water purity and clinical diagnosis

Chromatography plays an important role in many pharmaceutical industries and also in the chemical and food industry. Environmental testing laboratories generally wants to identify for very small quantities of contaminants such as PCBs in waste oil, and pesticides. The Environmental Protection Agency makes the method of chromatography to test drinking water and to monitor air quality. Pharmaceutical industries use this method both to prepare huge quantities of extremely pure materials, and also to analyze the purified compounds for trace contaminants. These separation techniques like chromatography gain importance in different kinds of companies, different departments like Fuel Industry, biotechnology, biochemical processes, and forensic science. Chromatography is used for quality analyses and checker in the food industry, by identifying and separating, analyzing additives, vitamins, preservatives, proteins, and amino acids. Chromatography like HPLC is used in DNA fingerprinting and bioinformatics

The method of separation is united type of unit procedure in most of the modern pharmaceutical, chemical and other process plants. Newer separation processes, like super critical fluid chromatography, paper chromatography hybridization assay chromatographic separation, etc., are gaining importance in modern days plants as novel separation processes. The Hyphenated Technique is increased and made advanced by uniting of a separation technique and an on-line spectroscopic detection technology. In this session latest update in the advancement of different hyphenated techniques, e.g., LC-FTIR, LC-NMR, CE-MS, GC-MS, Liquid Chromatography MS, etc. in the method of pre-isolation identification of crude extracts or fractions from different natural sources, separation and  identification of natural products, chemotaxonomic studies and DNA fingerprinting are included

The Chromatography industry is showing growth rapidly, with value expected to hit 228 billion dollars by 2016, up from 164 US billion dollars in the year of earlier 2010, growing annual growth of nearly 7%, according to a recent industrial marketing research report. Geographically, global separation technique market has been segmented into four areas namely, North America, some parts of European region, Asia-Pacific and Rest of the World. The global gas chromatography market, valued at $2,583.6 million in earlier 2014, is possibility to march up to $3,605.1 US million by 2019 at a CAGR of 6.9%.This global market report categorizes the market into 3 wide segments, like, Instrumentation, Accessories & Consumables, and Reagents

Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures. Thin-layer chromatography is performed on a sheet of an inert substrate such as glass, plastic, or aluminum foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminum oxide (alumina), or cellulose. This layer of adsorbent is known as the stationary phase.

Thin-layer chromatography can be used to monitor the progress of a reaction, identify compounds present in a given mixture, and determine the purity of a substance. Specific examples of these applications include: analyzing ceramists and fatty acids, detection of pesticides or insecticides in food and water, analyzing the dye composition of fibers in forensics, as saying the radiochemical purity of radiopharmaceuticals, or identification of medicinal plants and their constituents

Paper chromatography is an analytical method used to separate colored chemicals or substances. It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC).

A paper chromatography variant, two-dimensional chromatography, involves using two solvents and rotating the paper 90° in between. This is useful for separating complex mixtures of compounds having similar polarity, for example, amino acids. The setup has three components. The mobile phase is a solution that travels up the stationary phase, due to capillary action. The mobile phase is generally a mixture of non-polar organic solvent, while the stationary phase is polar inorganic solvent water. Here paper is used to support the stationary phase, water. Polar water molecules are held inside the void space of the cellulose network of the host paper. The difference between TLC and paper chromatography is that the stationary phase in TLC is a layer of adsorbent (usually silica gel, or aluminum oxide), and the stationary phase in paper chromatography is less absorbent paper