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A Seminar On GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple andandandand
A
Seminar
On
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple andandandand ApplicationApplicationApplicationApplication
DELIVEREDDELIVEREDDELIVEREDDELIVERED BY:BY:BY:BY:
UNDERUNDERUNDERUNDER GUIDANCEGUIDANCEGUIDANCEGUIDANCE BY:BY:BY:BY:
Deepak Kr. Verma
M,Sc,(Agri. Biochemistry)
ID No.7483/09
E-mail:rajadkv@rediffmail.com
Dr. P. S. Kendurkar (Prof. & Head)
Dr, Madhu Vajpeyi (Assoc. Prof.)
DepartmentDepartmentDepartmentDepartment OfOfOfOf AgriculturalAgriculturalAgriculturalAgricultural BiochemistryBiochemistryBiochemistryBiochemistry
CHANDRACHANDRACHANDRACHANDRA SHEKHARSHEKHARSHEKHARSHEKHAR AAZADAAZADAAZADAAZAD UNIVERSITYUNIVERSITYUNIVERSITYUNIVERSITY OFOFOFOF
AGRICULTUREAGRICULTUREAGRICULTUREAGRICULTURE &&&& TECHNOLOGYTECHNOLOGYTECHNOLOGYTECHNOLOGY
KANPURKANPUR-KANPURKANPUR--208002-208002208002208002
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
INTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTIONINTRODUCTION
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
ChromatographyChromatographyChromatographyChromatography
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MadeMadeMadeMade upupupup bybybyby
ChromaChromaChromaChroma ==== aaaa colorcolorcolorcolor
&&&&
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RussianRussianRussianRussian BotanistBotanist,BotanistBotanist,,,
MichaelMichaelMichaelMichael SemonavichSemonavichSemonavichSemonavich TswettTswettTswettTswett (L.T.(L.T.(L.T.(L.T. 18721872-18721872--1919)-1919)1919)1919)
InInInIn 1906190619061906
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ureureureure
wherewherewherewhere mixturemixturemixturemixture ofofofof differentdifferentdifferentdifferent coloredcoloredcoloredcolored pigmentspigmentspigmentspigments (such(such(such(such aaaassss chlorophyllchlorophyllchlorophyllchlorophyll &&&&
xanthophylls)xanthophylls)xanthophylls)xanthophylls) isisisis separatedseparatedseparatedseparated fromfromfromfrom eacheacheacheach other,other,other,other, meansmeansmeansmeans ooooriginallyriginallyriginallyriginally thethethethe
separatedseparatedseparatedseparated coloredcoloredcoloredcolored substancesubstancesubstancesubstance fromfromfromfrom thethethethe extractextractextractextract ofofofof thethethethe leaf.leaf.leaf.leaf. ByByByBy thisthisthisthis
methodmethodmethodmethod TswettTswettTswettTswett showedshowedshowedshowed thatthatthatthat thethethethe chlorophyllchlorophyllchlorophyllchlorophyll isisisis thethethethe mamamamadededede upupupup ofofofof twotwotwotwo
pigmentspigmentspigmentspigments andandandand itititit becomesbecomesbecomesbecomes moremoremoremore clearclearclearclear whenwhenwhenwhen thethethethe petrolepetrolepetrolepetroleumumumum etheretheretherether waswaswaswas
purepurepurepure inininin thethethethe packedpackedpackedpacked column.column.column.column.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
DEFINITIONDEFINITIONDEFINITIONDEFINITIONDEFINITIONDEFINITIONDEFINITIONDEFINITION
OfOfOfOfOfOfOfOf
CHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHY
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
CChCChhrhrroroomommamaatattotoogoggrgrraraapapphphhyhyyy ==== BBiBBiioioococchchheheememmimiiciccacaalalll TTeTTeececchchhnhnniniiqiqququueueesesss ==== CCoCCoomommpmppopoononneneenenntnttt oofoofff aaaa MMiMMiixixxtxttutuururrereee
ThroughThroughThroughThrough theirtheirtheirtheir differentdifferentdifferentdifferent migration.migration.migration.migration.
SeparatedSeparatedSeparatedSeparated
TheTheTheThe techniquetechniquetechniquetechnique inininin whichwhichwhichwhich thethethethe componentscomponentscomponentscomponents ofofofof thethethethe mixtumixtumixtumixturererere
areareareare causedcausedcausedcaused totototo migratemigratemigratemigrate atatatat differentdifferentdifferentdifferent ratesratesratesrates throughthroughthroughthrough onononon apparatusapparatusapparatusapparatus
whichwhichwhichwhich involveinvolveinvolveinvolve inininin thethethethe equilibriumequilibriumequilibriumequilibrium ofofofof thethethethe compoundscompoundscompoundscompounds bbbbetweenetweenetweenetween aaaa
stationarystationarystationarystationary andandandand aaaa mobilemobilemobilemobile phase.phase.phase.phase.
AAAA processprocessprocessprocess bybybyby whichwhichwhichwhich thethethethe componentcomponentcomponentcomponent ofofofof aaaa mixturemixturemixturemixture areareareare
separatedseparatedseparatedseparated bybybyby differentdifferentdifferentdifferent movementmovementmovementmovement throughthroughthroughthrough twotwotwotwo phasephasephasephase ssssystemystemystemystem inininin
whichwhichwhichwhich thethethethe movementmovementmovementmovement isisisis effectedeffectedeffectedeffected bybybyby aaaa mobilemobilemobilemobile phasephasephasephase ththththroughroughroughrough aaaa
stationarystationarystationarystationary phase.phase.phase.phase.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
TheTheTheThe techniquetechniquetechniquetechnique ofofofof separationseparationseparationseparation ofofofof substancessubstancessubstancessubstances accordingaccordingaccordingaccording totototo
theirtheirtheirtheir partitionpartitionpartitionpartition coefficientcoefficientcoefficientcoefficient orororor distributiondistributiondistributiondistribution coefficcoefficcoefficcoefficientientientient (K(K(K(Kdddd))))
belowbelowbelowbelow (i.e.(i.e.(i.e.(i.e. theirtheirtheirtheir relationrelationrelationrelation solubility)solubility)solubility)solubility) inininin twotwotwotwo impreimpreimpreimpressiblessiblessiblessible
phasephasephasephase
Here,Here,Here,Here,
(K(K(K(Kdddd)))) ==== ConcentrationConcentrationConcentrationConcentration ofofofof phasephasephasephase AAAA
ConcentrationConcentrationConcentrationConcentration ofofofof phasephasephasephase BBBB
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
MODESMODESMODESMODESMODESMODESMODESMODES
OfOfOfOfOfOfOfOf
CHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHY
ChromatographyChromatographyChromatographyChromatography separationseparationseparationseparation
maymaymaymay bebebebe achievedachievedachievedachieved bybybyby
twotwotwotwo basicbasicbasicbasic techniques,techniques,techniques,techniques,
theytheytheythey areareareare
1-Column chromatography
2-Thin-layer or Planer chromatography
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
PRINCIPLEPRINCIPLEPRINCIPLEPRINCIPLEPRINCIPLEPRINCIPLEPRINCIPLEPRINCIPLE
OFOFOFOFOFOFOFOF
CHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHY
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
1:-Different rates of migration of the component
of a mixture on the solvent system.
2:-Different rates of adsorption of the
components in the stationary phase.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
TypesTypesTypesTypesTypesTypesTypesTypes
OfOfOfOfOfOfOfOf
CHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHY
In all the chromatography techniques the phenomena
of the adsorption or partition is involved. In the
adsorption, the binding of compounds to the surface of
solid phase (stationary phase) takes place, where as in the
partition the relatively solubility of the compound in two
phase, result in the partition of the compounds in two
phase.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
1:-Adsorption chromatography (Compt. b/w S &L/G)
1:--Adsorption Column Chromatography (Compt. b/w S&G)
2:--Thin Layer Chromatography (Compt. b/w S&L)
3:--Ion-exchange Chromatography (Compt. b/w S&L)
2:-Partition chromato ra h
g
p
y
(Com t b/w L &L/G)
p .
1:-Paper Chromatography (Compt. b/w L&L)
2:--Thin Layer Chromatography (Compt. b/w L&L)
3:--Gel Filtration (Compt. b/w L&L)
4:--Gas Liquid Chromatography
Or
Vapor Phase Chromatography
(Compt. b/w L&G)
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
Gas-solid Chromatography (GSC)
Or
Gas liquid chromatography (GLC)
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
11-11--In-InInIn GSCGSCGSCGSC andandandand GLC,GLC,GLC,GLC, thethethethe separationseparationseparationseparation isisisis carriedcarriedcarriedcarried outoutoutout inininin aaaa tubulartubulartubulartubular
columncolumncolumncolumn mademademademade upupupup ofofofof aaaa glass,glass,glass,glass, metalmetalmetalmetal orororor telfon.telfon.telfon.telfon.
22-22--In-InInIn GSC,GSC,GSC,GSC, thethethethe adsorbentadsorbentadsorbentadsorbent areareareare thethethethe first,first,first,first, firstfirstfirstfirst finelyfinelyfinelyfinely dddivideddividedividedivided intointointointo
thethethethe finefinefinefine sizesizesizesize gradedgradedgradedgraded powderpowderpowderpowder andandandand thenthenthenthen fillfillfillfill inininin thethethethe cocococolumnlumnlumnlumn
,,,,
nonnon-nonnon--volatile-volatilevolatilevolatile liquid.liquid.liquid.liquid.
33-33--GSC-GSCGSCGSC isisisis basedbasedbasedbased uponuponuponupon thethethethe selectivityselectivityselectivityselectivity adsorptionadsorptionadsorptionadsorption onononon aaaa sssolidsolidolidolid whilewhilewhilewhile
thatthatthatthat inininin GLCGLCGLCGLC isisisis basedbasedbasedbased uponuponuponupon thethethethe partitionpartitionpartitionpartition betweenbetweenbetweenbetween thethethethe gasgasgasgas
andandandand immobileimmobileimmobileimmobile (stationary(stationary(stationary(stationary phase)phase)phase)phase) liquidliquidliquidliquid phase.phase.phase.phase.
44-44--The-TheTheThe basicbasicbasicbasic componentcomponentcomponentcomponent ofofofof thethethethe GSCGSCGSCGSC andandandand GLCGLCGLCGLC typetypetypetype ofofofof
chromatographychromatographychromatographychromatography areareareare thethethethe samesamesamesame andandandand shownshownshownshown inininin thethethethe figurefigurefigurefigure
whilewhilewhilewhile thatthatthatthat inininin GLCGLCGLCGLC thethethethe columncolumncolumncolumn wallwallwallwall isisisis eithereithereithereither coatedcoatedcoatedcoated withwithwithwith
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
G.L.C.G.L.C.G.L.C.G.L.C.G.L.C.G.L.C.G.L.C.G.L.C.
AAAA gasgasgasgas liquidliquidliquidliquid chromatographychromatographychromatographychromatography isisisis thethethethe analysisanalysisanalysisanalysis ofofofof thethethethe organicorganicorganicorganic
metabolitesmetabolitesmetabolitesmetabolites hashashashas beenbeenbeenbeen usedusedusedused widelywidelywidelywidely forforforfor thethethethe analysisanalysisanalysisanalysis ooooffff thethethethe fat,fat,fat,fat, fattyfattyfattyfatty
acids,acids,acids,acids, pesticidespesticidespesticidespesticides andandandand severalseveralseveralseveral volatilevolatilevolatilevolatile substances.substances.substances.substances. ExtensiveExtensiveExtensiveExtensive studiesstudiesstudiesstudies
havehavehavehave beenbeenbeenbeen mademademademade possiblepossiblepossiblepossible onononon thethethethe organicorganicorganicorganic volatilevolatilevolatilevolatile inininin foodfoodfoodfood andandandand
fermentedfermentedfermentedfermented beveragebeveragebeveragebeverage wherewherewherewhere flavorflavorflavorflavor andandandand aromaaromaaromaaroma areareareare ofofofof alalalalmostmostmostmost
imimimim ortanceortanceortanceortance
InInInIn generalgeneralgeneralgeneral considerationconsiderationconsiderationconsideration ofofofof GLCGLCGLCGLC techniques,techniques,techniques,techniques, thethethethe colcolcolcolumnumnumnumn isisisis
packedpackedpackedpacked withwithwithwith aaaa purepurepurepure inertinertinertinert solidsolidsolidsolid coatedcoatedcoatedcoated withwithwithwith aaaa thinthinthinthin llllayerayerayerayer ofofofof aaaa
nonvolatilenonvolatilenonvolatilenonvolatile liquidliquidliquidliquid asasasas thethethethe stationarystationarystationarystationary phase.phase.phase.phase. TheTheTheThe comcomcomcomponentsponentsponentsponents ofofofof thethethethe
mixturemixturemixturemixture areareareare separatedseparatedseparatedseparated bybybyby beingbeingbeingbeing partitionedpartitionedpartitionedpartitioned b/wb/wb/wb/w thisthisthisthis phasephasephasephase andandandand thethethethe
mobilemobilemobilemobile phase.phase.phase.phase.
pppp
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC exploits differences in the partition coefficient
between a stationary liquid phase and a mobile gas phase of
volatilized analyst as they are carried out through the column
by the mobile gas phase means the technique is based upon
the partitioning compounds between a liquid and a gas
phase.
Therefore, it’s used to confine the analyst that are
volatile but thermally stable, ’The’The’The’The partitionpartitionpartitionpartition coefficientcoefficientcoefficientcoefficient areareareare inverselyinverselyinverselyinversely
proportionalproportionalproportionalproportional totototo thethethethe volatilityvolatilityvolatilityvolatility ofofofof thethethethe analyst’.analyst’.analyst’.analyst’.
The temperature of the column is raised to 50-300 ºC to
facilitate analyst volatilization. There is optimum flow arte of
the mobile phase for maximum column efficiency (minimum
plate height, H)
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
The stationary phase is consists of…
►A high boiling points liquid materials
such as silicon, grease or wax
The materials is either coated onto the
internal wall of the column or supported on
inert granular solid and packed into the
column.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
1-A column house in an oven that can be
temperature programmed
2-A Sample Inlet Points
3- A Carrier Gas Supply & Control
4- A Detector
5- An Amplifier
6- Data Record System
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
Filters/Traps
Data system
H
RESET
Regulators
Syringe/Sampler
Inlets
gas
Detectors
system
inlet
Column
column
Gas Carrier
detector
Hydrogen
data
system
Air
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
There are two type of column used in the GLC, they are …
1- Packed Conventional Column
2- Capillary (open tubular) Column
Packed Conventional Column- These consist of coil glass or stainless
steel column 1-2 mm long and 2-4 mm internal diameter. They are
packed with stationary phase coated onto on inert silica support.
Capillary (open tubular) Column-These are made by high quality
fazed quartz and or 10-100 m and 0.1-1.0 mm diameter.
This type of column are further divided in two type, they are
1-Wall-coated Open Tubular (WCOT)
2-Support-coated Open Tubular (SCOT)
also called as Porous Layer Open Tubular (PLOT)
The operating temperature for all type of column must be
compatible with stationary phase chosen for use column temperature is
controlled to ±0.1°C.
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
DETECTORDETECTORDETECTORDETECTORDETECTORDETECTORDETECTORDETECTOR
UsedUsedUsedUsedUsedUsedUsedUsed
InInInInInInInIn
GLCGLCGLCGLCGLCGLCGLCGLC
Several types of detectors are in common used in the
conjunction with gas-liquid chromatography system…
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
1- Flame Ionization Detector (FID)
2- Nitrogen Phosphorus Detector
(NPD)
3- Electron Capture Detector (NPD)
4- Flame Photometric Detector (ECD)
5- Rapid Scanning Furriers
Transforms Infrared Detector
6- Mass Spectrometer Detector (MSD)
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
APPLICATIONAPPLICATIONAPPLICATIONAPPLICATIONAPPLICATIONAPPLICATIONAPPLICATIONAPPLICATION
OfOfOfOfOfOfOfOf
CHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHYCHROMATOGRAPHY
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
GLC:GLC:GLC:GLC: PrinciplePrinciplePrinciplePrinciple &&&& ApplicationApplicationApplicationApplication
11-11--GLC-GLCGLCGLC isisisis veryveryveryvery usefulusefulusefuluseful forforforfor thethethethe analysisanalysisanalysisanalysis ofofofof complexcomplexcomplexcomplex mixtmixtmixtmixture.ure.ure.ure.
22-22--GLC-GLCGLCGLC isisisis widelywidelywidelywidely usedusedusedused forforforfor thethethethe qualitativequalitativequalitativequalitative andandandand quantitaquantitaquantitaquantitativetivetivetive analysisanalysisanalysisanalysis
ofofofof thethethethe largelargelargelarge numbernumbernumbernumber ofofofof aaaa lowlowlowlow polaritypolaritypolaritypolarity compoundscompoundscompoundscompounds be4be4be4be4cousecousecousecouse itititit hashashashas
highhighhighhigh sensitivity,sensitivity,sensitivity,sensitivity, andandandand speedspeedspeedspeed o0fo0fo0fo0f resolutions.resolutions.resolutions.resolutions.
33-33--Analytically,-Analytically,Analytically,Analytically, veryveryveryvery powerfulpowerfulpowerfulpowerful techniquetechniquetechniquetechnique whenwhenwhenwhen coupledcoupledcoupledcoupled totototo massmassmassmass
spectrometryspectrometryspectrometryspectrometry
44-44--Use-UseUseUse forforforfor thethethethe separationseparationseparationseparation ofofofof volatilevolatilevolatilevolatile substancessubstancessubstancessubstances
55-55--In-InInIn thethethethe clinicalclinicalclinicalclinical orororor medicalmedicalmedicalmedical chemistry,chemistry,chemistry,chemistry, itititit isisisis usedusedusedused fofofoforrrr thethethethe separationseparationseparationseparation
ofofofof plasma,plasma,plasma,plasma, lipids,lipids,lipids,lipids, blood,blood,blood,blood, alcohol,alcohol,alcohol,alcohol, urinaryurinaryurinaryurinary steroidsteroidsteroidsteroid etcetcetcetc
DeepakDeepakDeepakDeepak Kr.Kr.Kr.Kr. Verma,Verma,Verma,Verma, 2010201020102010
TechniquesTechniquesTechniquesTechniques inininin BiochemistryBiochemistryBiochemistryBiochemistry ABC-504
BibliographyBibliographyBibliographyBibliography Lehninger, Principle Of Biochemistry by Nelson David L. & Cox
BibliographyBibliographyBibliographyBibliography
Lehninger, Principle Of Biochemistry by Nelson David L. & Cox Micheal M,
Macmillan, Worth Publications
Principles and Techniques of Biochemistry & Molecular Biology
by Keith Wilson & John Walker
Edition Sixth 2005, Cambridge University Press
Practical Biochemistry, Principles and Techniques by Keith Wilson & John
Walker
Edition Fifth 2000, Cambridge University Press
Modern Practices of Gas Chromatography by Grob R.L.
Edition 3rd 1995, Wiley Intersciences, New York
A Biologist Guide to Principles & Techniques of Practical Biochemistry
by Wilson K., Gouleling KH (Editors)
Edition 3rd 1986, Edword Arnold (Publisher’s) Ltd. London
Fundamental of Biochemistry by Jain J.L., Jain Sanjay, Jain Nitin
Edition 2008, S.Chand & Company Ltd
Text Books of Plant Physiology and Biochemistry by Verma S.K.
Edition 2005, S.Chand & Company Ltd
Biochemistry by Fatima, Narayan & Meyya R.P.
Edition Fourth 2002, Saras Publications
Web Links www.biochem.orizona.edu www.biocore.com
Web Links
www.biochem.orizona.edu
www.biocore.com