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 what is enzymes, Explain what is Enzymes? Enzymes are organic...

Explain what is Enzymes? Enzymes are organic substances that speed up, or catalyze, a chemical reaction. At a given temperature, molecules have varying amounts of energy, and

How does photoperiodism affect the flowering of some plants, How does photo...

How does photoperiodism affect the flowering of some plants? The Flowering is an easy and typical to observe example of photoperiodism. Most of flowering plants flower only dur

What is p-k reaction, The response produced when an allergen is injected in...

The response produced when an allergen is injected into an individual, who is sensitive is known as P-K reaction.

Why do some physicians advise patients, Why do some physicians advise patie...

Why do some physicians advise patients with a bladder infection to drink a lot of cranberry juice? Answer Cranberry juice contains chemicals that kill viruses and bacteria. The inc

Biological diversity protection, Q. How can a great biological diversity pr...

Q. How can a great biological diversity protect an ecosystem from environmental damage? Why are less biodiverse ecosystems at risk of suffering deep biological harm if submitted to

Plants, In school laboratory, what is usually regarded as evidence that pho...

In school laboratory, what is usually regarded as evidence that photosynthesis has occurred in a plant

Zoonoses disease-leptospirosis, Leptospirosis Leptospirosis is a gener...

Leptospirosis Leptospirosis is a general term that denotes all infections of man and animals caused by spirochaetes of the genus Leptospira. At one time, the infection had bee

How to calculate the net protein utilization (npu), How to calculate the Ne...

How to calculate the Net Protein Utilization (NPU)? Mitchell (1922) introduced the term 'Net Utilization of Dietary Protein' which is a product of digestibility coefficient and

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