thuyet trinh phan tich 1

8/8/2019 Thuyet Trinh Phan Tich 1

1/22

OutlineDefinition of ChromatographyHistory of Chromatography

Chromatography terms

Classification of Chromatography

Chromatograph

Applications of Chromatography


2/22

Definition

Is the collective term for a family of laboratory techniques

for the separation of mixtures

Involves passing a mixture which contains the analyte

through a stationary phase, which separates it from other

molecules in the mixture and allows it to be isolated

SeparateAnalyze Identify

Purify

QuantifyComponentsMixture


3/22


4/22

Chromatography Terms

The analyte is the substance that is to be separated

during chromatography

Stationary phase: Phase that stays in place inside thecolumn. Can be a particular solid or gel-based packing (LC)

or a highly viscous liquid coated on the inside of the

column (GC)

Mobile phase: Solvent moving through the column,

either a liquid in LC or gas in GC


5/22

Eluent: Fluid entering a column

Eluate: Fluid exiting the column

Elution: The process of passing the mobile phase

through the column.

Chromatogram: Graph showing detector response

as a function of a time.

Flow rate: How much mobile phase passed /

minute (ml/min).

Linear velocity: Distance passed by mobile phase

per 1min in the column (cm/min)


6/22

Chromatography

Column Planar Thin layer

Paper

physical state ofmobile

phase

GasLiquid

Supercritical

fluid

Separation mechanism

Size

exclusion

Ion

exchange

Classification


7/22

Column Chromatography

The stationary phase (column

packing) in the column is very polar

One starts will a less polar solvent to

remove

the less polar compounds, and then

you slowly

Increase the polarity of the solvent to

remove the more polar compounds

Figure 1. column chrotography .

solvent

column

Bands of

separatedconsistuents


8/22

Thin LayerChromatography

Here the mobile phase is a liquid

Flowing past a thin layer of powder on a solid

support

Substances that are less attracted to the solid or are

more soluble in the liquid move faster

And so move further up the plate by the time that the

process has been stopped by taking the plate out of

the liquid. - larger Rf


9/22

Figure 2. principles in TLC.


10/22


11/22

PaperChromatography

Figure 3. Paper chromatography jar


12/22


13/22

Size exclusion ChromatographyGEL-PERMEATION CHROMATOGRAPHY

Gel-Permeation Chromatography is a mechanical sorting of

molecules based on the size of the molecules in solution.

Small molecules are able to permeate more pores and are,

therefore, retained longer than large molecules.


14/22

Separation in Ion-exchange Chromatography is based on

the competition of different ionic compounds of the sample

for the active sites on the ion-exchange resin (column-

packing).

Ion- exchange Chromatography


15/22

Liquid chromatography (LC) is a separation technique in

which the mobile phase is a liquid. Liquid

chromatography can be carried out either in a column or

a plane

Liquid Chromatography

Present day, use high

performance liquid

chromatography (HPLC)

Figure 4 . HPLC 1525


16/22

Gas Chromatography

Principals of SeparationColumn is selected, packed with Liquid Phase, and installed.

Sample injected with microliter syringe into the injection

port where it is vaporized and mixed into the Carrier Gas

stream (helium, nitrogen, argon).

Sample vapor becomes partitioned between Moving Gas

Phase and Stationary Liquid Phase

The time the different compounds in the sample spend in

the Vapor Phase is a function of their Vapor Pressure.

The more volatile (Low Boiling Point / Higher Vapor

Pressure) compounds arrive at the end of the column first

and pass into the detector


17/22

Chromatograph

Figure 5. chromatograph


18/22

Figure 6. chromatographic peak is fronting

Figure 7. chromatographic peak is tailing


19/22

Applications

Figure 6. Gas chromatographyFigure 5. HPLC


20/22

Chromatography is used by scientists to:

Analyze examine a mixture, its components,and their relations to one another

Identify determine the identity of a mixture orcomponents based on known components

Purify separate components in order to isolate

one of interest for further study

Quantify determine the amount of the a mixtureand/or the components present in the samplecxxc


21/22

Pharmaceutical Company determine amount ofeach chemical found in new product

Hospital detect blood or alcohol levels in apatients blood stream

Law Enforcement to compare a sample found at

a crime scene to samples from suspects

Environmental Agency determine the level ofpollutants in the water supply

Manufacturing Plant to purify a chemical neededto make a product

So on


22/22

References

http://en.wikipedia.org/wiki/Chromatography

www.chromatography-online.org

www.docjax.com/.../index.shtml?...thin%20layer%20 chomatography

www.ebookpdf.net/__chomatography-free-book_ebook_.html

www.waters.com/.../library.htm...

thuyet trinh phan tich 1

Documents