Define about the column chromatography, Biology

Assignment Help:

Define about the Column chromatography?

In column chromatography, a tube is filled with the material constituting the stationary phase, plus a solvent (mobile phase). The general adsorbent (stationary phase) used are alumina, silica gel, activated carbon, calcium phosphate and hydroxyapatite. The substances to be separated are dissolved in the smallest possible volume of a suitable solvent and applied on the top of the stationary phase and allowed to enter the column. The chromatogram is then developed by flowing a solvent (the mobile phase) through the column. As different substances move through the column, they separate and appear in the effluent when particular volumes of liquid have passed through the column. The liquid leaving the column (the eluent) is usually collected as discrete fractions, using an automatic collector.

The separated components are then identified by testing aliquots of each fraction by one or a combination of the following methods viz. colorimetry, UV absorption, fluorimetry, scintillation counting, refractive index, spectral diode array system, electrochemical detection, radioimmunoassay and enzyme immunoassay. When automatic fraction collector is used the recorder automatically draws each peak and the area of each peak is proportional to the amount of sample component present in it. The simplest form of column chromatography is adsorption chromatography. Separation of components by this method depends upon differences both in their degree of adsorption by the adsorbent and solubility in the solvent used for separation. These physiochemical factors are governed by the molecular structure of the compound.


Related Discussions:- Define about the column chromatography

Explain carbon and oxygen aspect of using phase rule, Explain carbon and ox...

Explain carbon and oxygen aspect of using phase rule? Consider a system containing solid carbon (graphite) and a gaseous mixture of O 2 , CO, and CO 2 . There are four species

Two traits are involved in a genetic cross, In his experiments, Mendel note...

In his experiments, Mendel noted that when two traits are involved in a genetic cross, they are inherited independently of each other. The reason for this is that A. genes on the s

Where does the water resorbed after glomerular filtration, Q. Where does mo...

Q. Where does most of the water resorbed after glomerular filtration go? What are the other substances resorbed by the nephron tubules? Only 0.5 to 1% of the glomerular filtrat

Why is genetic homozygosity dangerous, Why is genetic homozygosity dangerou...

Why is genetic homozygosity dangerous to a nearly extinct species? Genetic homozygosity leaves no variation for natural selection to act on. Thus, a new disease could wipe

Plate tectonic theory, Plate Tectonic Is a Theory of Geology Which describe...

Plate Tectonic Is a Theory of Geology Which describes the large scale motion of earth's lithosphere. The theory builds on the older theory of continental drift from the first half

Discuss about human ears, Ears We have a pair of ears. Each ear has thr...

Ears We have a pair of ears. Each ear has three parts - external, middle and inner ear. The function of the middle ear is transmission of sound waves from external to inner ear

The mirrows reach or grayling zone, The mirrows reach or grayling zone ...

The mirrows reach or grayling zone This zone has a less steep gradient, than the trout beck. The river here still flows swiftly, though the current is not fast and conditions

Metabolic reactions in the hmp pathway, Metabolic Reactions  in the HMP Pa...

Metabolic Reactions  in the HMP Pathway The hexose monophosphate pathway is responsible for  the generation of a substantialfraction of the cytoplasmic NADPH required  for bios

Give three examples of reflex actions, Give three examples of reflex action...

Give three examples of reflex actions. Examples of reflex actions are alter in size of the pupil of the eye in response to light intensity, blinking in response to foreign par

Wetlands and estuaries, Wetlands and estuaries are transitional biomes. Lan...

Wetlands and estuaries are transitional biomes. Land that remains flooded either part of the year or permanently with fresh or salt water is known as wetland. The internationally a

Write Your Message!

Captcha
Free Assignment Quote

Assured A++ Grade

Get guaranteed satisfaction & time on delivery in every assignment order you paid with us! We ensure premium quality solution document along with free turntin report!

All rights reserved! Copyrights ©2019-2020 ExpertsMind IT Educational Pvt Ltd