AN INTRODUCTION TO CHROMATOGRAPHY

ByLubna Ahmed

Shantel HernandezKeisha Napper

Dayimwende OuedraogoChristine Pennington

Sunny Schemery

WHAT IS CHROMATOGRAPHY?

Chromatography is a method used for the separation of a mixture of compounds into its individual components

The purpose is to detect or quantitate a particular component to be assayed in a pure form.

Chromatography has 2 phase: Stationary Phase: the material in the column for which

the components to be separated having varying affinities.

Mobile Phase: comprised of solvents into which the sample is injected. Referred to as “carrier fluid” due to the solvent and the sample flow through the column together.

How does Chromatography work?

The stationary phase stays fixed in place while the mobile phase moves the components of the mixture through medium or column being used.

Stationary phase acts as a constraint, slowing down the movement of some components in the mobile phase.

Different factors of a component affect the affinities for the stationary phase, which makes components move faster or slower facilitating the separation of the components in the mixture.

Equipment used in Chromatography

The Column: used in most application of chromatography. It is were the actual separation takes place.

Usually made of glass or metal tube of with enough strength to withstand the pressure that may be applied across it.

The column contains the stationary phase

The mobile phase runs through the column and is absorbed onto the stationary phase.

The column is either a packed bed or open tubular

Basic operation in Chromatography

Feed injection into the mobile phase flows through the system by the action of a pump.

Separation in the Column- sample flows through and the different components will absorb to the stationary phase to different degrees. A strong attraction will move slower than components with a weak attraction. This is how the separation occurs.

Elution from Column- when sample is flushed/displaced from stationary phase the different components will elute at different times. Weak attraction will come out first while strongly attracted components will be last.

Detection- the different components are collected as they come out of the column. A detector analyzes the stream by measuring a property related to concentration and characteristics of chemical composition.

Column Chromatography

Applications of Chromatography

Used to separate different particles in a solution for analyzing.

Used in chemistry as a method of isolating certain particles for testing

Size exclusion/ Gel Chromatography

Used in environmental science, food, cosmetics, forensic etc.

Applications of Chromatography:Thin Layer

Used to identify drugs present in blood or Thin Layer Chromatography

urine. Used with:

Athletes Job applicants People on

probation

Animation of Size Exclusion Chromatography

Click/Copy link below http://

www.separations.us.tosohbioscience.com/ServiceSupport/TechSupport/ResourceCenter/PrinciplesofChromatography/SizeExclusion

Citation List Carrier, R. (n.d.). Chromatography. Retrieved from

http://www.rpi.edu/dept/chem-eng/Biotech-Environ/CHROMO/chromintro.html

Chromatography [Web Graphic]. Retrieved from http://www.m2c3.com/chemistry/VLI/M4_Topic2/la_16_07.jpg

Chromatography. (2013). Retrieved from http://science.howstuffworks.com/chromatography-info.htm

Column Chromatography [Web Graphic]. Retrieved from wdict.net/word/Column chromatography

High Performance Liquid Chromatography (HPLC). (2008). In S. Bell, Facts on File Science Library. Encyclopedia of Forensic Science, Rev. ed. (pp. 188-189). New York: Facts on File. Retrieved from http://go.galegroup.com/ps/i.do?id=GALE%7CCX3226900320&v=2.1&u=la32265&it=r&p=GPS&sw=w

Size Exclusion Chromatography [Web Graphic]. Retrieved from http://bricker.tcnj.edu/tech/BIOL311chromatography.htm

Thin Layer Chromatography [Web Graphic]. Retrieved from http://withfriendship.com/user/svaruna/thin-layer-chromatography.php

Testing athletes for drug uses. (2013). Retrieved from http://entertainment.howstuffworks.com/athletic-drug-test8.htm

Turgeon, M. L. (2011). Linne & ringsrud's clinical laboratory science: The basics and routine techniques. (6 ed., p. 155). Mobsy publishing.

(2013). World Book, Inc. Retrieved from http://www.worldbookonline.com/pl/infofinder/article?id=ar113900

Thin Layer Chromatography [Web Graphic]. Retrieved from http://withfriendship.com/user/svaruna/thin-layer-chromatography.php