Chromatography Flashcards
principle of chromatography
adsorption or partition
column chromatography
stationary phase is solid or liquid supported by a solid
thin layer chromatography
used to separate mixtures, check purity of a mixture and to monitor the progress of reaction
paper chromatography
liquid liquid chromatography can also be carried out on a strip of paper. thin liquid film of solvent held on the paper will act as a stationary phase.
adsorption CC
column of solid is used as adsorbent
partition CC
when solid column is acting only as a support to the liquid adsorbent
chromatogram
a banded column of adsorbed constituents
elution and eluent
dissolving out of component s form adsorbent is called elution and the solvent is called eluent
applications of CC
1) quantitative separation
2) purification
3) concentration
4) identification
chromatoplate
glass plate with thin layer of stationary phase
how do you get the result if the components are colourless in TLC
the dried plate is sprayed with a suitable reagent to make the spot coloured
Rf value
the ratio of distance travelled by a component to the distance travelled by solvent front
advantages of TLC
1) separation of minute quantities
2) sensitive and sharp separation
3) high speed of separation
4) wide choice of stationary phases
applications of TLC
1) detection of purity and identification
2) pharmaceutical, food and cosmetic industry
3) biochem analysis
HPLC
Modern method and powerful tool for the separation, purification and identification of various non volatile high molecular weight organic compounds and natural products
advantages of HPLC
1) high speed separation
2) excellent column separation
3) solvent consumption is minimum
4) sensitive and accurate
5) can be done in ambient temp
2 types of detectors in HPLC
1) bulk property
2) solute property
2 ways in which elution can be done
1) isocratic done by 1 solvent of constant composition
2) gradient done by 2 or more solvents differing in polarities
applications of HPLC
1) quality control
2) process control
3) forensic analysis
4) environmental monitoring
5) clinical testing
6) biomed research work
gas chromatography
for separation and analysis of gaseous mixture and volatile organic compounds
applications of gas chromatography
1) analysis of natural gas, gasoline plant samples, refinery gases, synthetic rubber and plastic intermediates, steroidal hormones
2) separation of close boiling liquids
3) determination of ethyl alcohol content in blood using glycol column
4) multicomponent analysis
5) steroidal hormones
analyte
substance to be separated during chromatography
bonded phase
stationary phase that is covalently bonded to the support particles or to the inside wall of the column tubing
chromatogram
visual output of chromatograph
