Analytical Chemistry Quiz Session 7-8

Gas Chromatography

As an analytical technique to separate chemical components in a sample mixture and then after that, it will detect the component to determine the compounds and how much compounds

Barbital(8.6pH)

Buffer used in electrophoresis

acetonitrile, methanol, ethanol, isopropanol and water

Commonly used solvents in HPLC?

Cellulose acetate Agarose gel Polyacrylamide gel

Commonly used supporting media of electrophoresis

Electrical power Support medium Buffer Sample and detecting system

Components of electrophoresis

Nitrogen, Helium, Hydrogen, Argon.

Examples of Mobile phase of GS

Net electric charge of the molecule Size & shape of the molecule Electric field strength Nature of supporting medium Temperature of operation

Factors that affect the rate of migration

boiling point, solubility or adsorption.

GC components migrate at different rates due to differences

steroids, barbiturate, blood, alcohol and lipids.

GC use in separating the?

organic materials

Gas chromatography use?

clinical and forensic work

HPCE also useful in?

Qualitative and quantitative characterization of biologically active materials

HPCE is used for?

adsorption, size or charge

HPLC components migrate through the column at different rates due to

hormones, lipids, carbohydrates, fractionation of drugs and proteins.

HPLC used for?

volatile and thermally stable

In gas chromatography sample must be?

below 400 celcius

In gas chromatography, is thermally stable at?

Chromatography

Involves of separation of soluble components in a solution.

Inorganic (mixtures of metals with similar chemical characteristics)

Ion-exchange chromatography use in?

Thin Layer Chromatography (TLC)

It is a type of planar chromatography (paper chromatography)

solvent polarity

Mobile phase of TLC

Gas

Mobile phase of gas chromatography

Size Exclusion Chromatography (SEC)

Molecules that differ in size can be separated by passing the sample solution through a stationary phase.

2-5%

Relative precision of gas chromatography

Agarose gel

Separates by electrical charge and doesn't bind to proteins.

Cellulose acetate

Separates by molecular size. The most commonly used for serum proteins

Polyacrylamide gel

Separates on the basis of charge and molecular size. It is used to study isoenzymes.

Size Exclusion Chromatography (SEC) application

Separation and desalting of high-molecular weight materials; determination of molecular weights

HPCE principles

Separation mainly of charged materials by differential migration across a surface or through a column in an applied potential gradient

gas-solid chromatography

Separation occurs based in differences in absorption at solid phase surfaces

gas-liquid chromatography

Separation occurs by differences in solute partitioning between the gaseous mobile phase and the liquid stationary phase.

ion exchange chromatography principles

Separation of ionic materials in microgram to gram quantities by passage of a solution through a column or across a surface consisting of a porous polymeric resin incorporating exchangeable ions.

Size Exclusion Chromatography (SEC) principles

Separation of materials according to molecular size and shape by passage of a solution through a column or across a surface consisting of a polymeric gel.

Thin Layer Chromatography (TLC) principles

Separation of mixtures in microgram quantities by movement of a solvent across a flat surface

Gas Chromatography principles

Separation of mixtures in microgram quantities by passage of the vaporized sample in a gas stream through a column containing a stationary liquid or solid phase

HPLC (high performance liquid chromatography) principles

Separation of mixtures in microgram to gram quantities by passage of the sample through a column containing a stationary solid by means of a pressurized flow of a liquid mobile phase

High Performance Liquid Chromatography (HPLC)

Separation of various hemoglobin associated with a disease.

High boiling liquid and sorption process

Stationary phase of Gas-liquid chromatography

solid and adsorption

Stationary phase of Gas-solid chromatograpy

insoluble but porous resinous material

Stationary phase of Ion-exchange chromatography

Matrix

Support medium of electrophoresis

solubility, adsorption, size or charge

TLC components migrate at different rates due to differences:

qualitative purposes and for both organic and inorganic materials

TLC is widely used in

mobile phase stationary phase the column holding the SP The separated component

The basic components of chromatography

urine drug screening

Thin layer chromatography is clinically used for

ion-exchange or gelpermeation materials

Thin layers are sometimes made from?

ELECTROPHORESIS

This is defined as the movement of charged molecules in a liquid medium when an electric field is applied

CAPILLARY ELECTROPHORESIS

This is used for separation, quantitation and determination of molecular weight of proteins and peptides.

ion exchange chromatography

This is used for the separation of amino acids, proteins and nucleic acids.

gas-solid chromatography, Gas-liquid chromatography

Type of gas chromatography

gas chromatography, high performance liquid chromatography, thin-layer chromatography, Ion-exchange chromatography, Size exclusion chromatography

Types of chromatography

HPLC (high performance liquid chromatography) application

Used largely for the separation of non-volatile substances including ionic and polymeric samples

Electroosmotic Flow (EOF)

also utilizes nanoliter quantities of specimens.

Ion exchange separation

are limited to samples containing ionized or partially ionized solutes

Solid samples

can be introduced as a solution or in a sealed glass ampoule which is crushed in the gas stream by means of a gas-tight plunger

stationary phase

can be silica gel or plastic sheet. (Solid or liquid). Where mobile phase flows

0.5-3%

capillary methods percent of HPCE

for both qualitative and quantitative analysis

gas chromatography id useful for

MOBILE PHASE

inert gas or a liquid and carries the complex mixture or the sample

High-performance capillary electrophoresis (HPCE)

is a comparatively recent development that is proving to be a very powerful separation technique of growing importance (vide infra

ELECTROPHORESIS

it is used clinically to separate and identify proteins, including serum, urine and cerebrospinal fluid (CSF) proteins, isoenzymes, and so on.

size-exculsion chromatography

it is used for high-molecular weight material.

Thin Layer Chromatography (TLC) application

it is useful for checks on purity, to monitor reactions and production processes and to characterize complex materials

Electrophoresis

it separates proteins(are negatively charge) depends on electric charge density

Liquid

mobile phase of HPLC

5-10%.

quantitative precision only moderate of TLC is?

Chromatography

refers to the group of techniques used to separate complex mixtures on the basis of different physical interaction

Electroosmotic Flow (EOF)

regulates the speed at which solutes move through the capillary

ion-exchange or exclusion.

sorption process of TLC

Silica gel or alumina

stationary phase of HPLC

porous cross-linked polymeric gel

stationary phase of SEC

Thin Layer Chromatography (TLC)

the separation process occurs on a flat essentially two-dimensional surface

Tswett in 1903

the word chromatography was used by...

Chromatography

to describe the separation of plant pigments by percolating a petroleum-ether extract through a glass column packed with powdered calcium carbonate.

5-20%

traditional methods percent of HPCE