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

Functions of colour in foods, Q. Functions of Colour in Foods? Let us n...

Q. Functions of Colour in Foods? Let us now look at the different functions of colour in details, which are enumerated herewith: 1. The maturity of many fruits and vegetable

Bty100, 1. (a) Which physicochemical properties need to be regulated by the...

1. (a) Which physicochemical properties need to be regulated by the cell? Define chemical equilibrium and discuss how it is maintained by the cell. (b) Differentiate between facili

Amoeboid tapetum - tapetum, Amoeboid Tapetum - Tapetum It is also know...

Amoeboid Tapetum - Tapetum It is also known as invasive or periplus modial tapetum. This type of tapetum is more prevalent in the monocotyledons (Arum) than in the dicotyledon

How is energy transferred along a food chain?, How is energy transferred al...

How is energy transferred along a food chain? The energy flux beside a food chain is always unidirectional, from the producers to the decomposers.

Plant responses to light-dark cycles, Plant Responses to Light-Dark Cycles ...

Plant Responses to Light-Dark Cycles Based on their requirement for day length (number of hours of light) for flowering plants have been grouped under three major categories,

Illustrate stents, Q. Illustrate Stents? Stents are metallic scaffolds ...

Q. Illustrate Stents? Stents are metallic scaffolds that are deployed within a diseased segment of a coronary artery to establish and then maintain a widely patent lumen. Stent

Define correlation between viscosity and solubility, Correlation between vi...

Correlation between viscosity and solubility Viscosity and consistency of protein systems are the important functional properties in fluid foods, such as  beverages, soups, sau

Salivary analyse works on foods, Explain how salivary amylase works on food...

Explain how salivary amylase works on foods like crackers

Annelida, why is the phylum annelida in trouble?

why is the phylum annelida in trouble?

Evolutionary changes in humans, Evolutionary changes that occurred in human...

Evolutionary changes that occurred in humans are - Development of prominent chin. Increase in cranial capacity. Reduction of brow-ridges. Development of speech. Developme

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