Sci 124 New Lab Chromatographic Separation Methods
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CHROMATOGRAPHIC SEPARATION METHODS
SEPARATION BY THIN-LAYER CHROMATOGRAPHY (TLC)&
REVERSE-PHASE
1,529,560 New Cancer Cases in 2010569,490 Cancer Deaths in 2010
Curcumin Prevents Cancer, Heart Disease, Alzheimer’s Disease and Obesity.
CURCUMIN, ANYONE ?CURCUMIN, ANYONE ?
Cancer CellCancer Cell
PurposePurpose::
ISOLATION OF THE YELLOW PIGMENT ISOLATION OF THE YELLOW PIGMENT CURCUMIN FOUND IN THE SPICE CURCUMIN FOUND IN THE SPICE TUMERICTUMERICBY THINBY THIN--LAYER CHROMATOGRAPHY LAYER CHROMATOGRAPHY (TLC) METHOD & USING A POLAR (TLC) METHOD & USING A POLAR
ORGANIC SOLVENT.ORGANIC SOLVENT.CurcuminCurcumin
Turmeric RootTurmeric Root
TUMERICTUMERIC COMES FROM THE COMES FROM THE CURCUMA CURCUMA LONGALONGAPLANT (GINGERPLANT (GINGER--LIKE) FOUND IN LIKE) FOUND IN ASIAASIA..
PART 1PART 1: : Thin Layer Chromatography Analysis of CURCUMIN in TurmericThin Layer Chromatography Analysis of CURCUMIN in Turmeric
Cell Cell MenbraneMenbrane: : PhospholipidPhospholipid BiBilayerlayer
Step #1: Heating Reflux Step #1: Heating Reflux
60 G OF STORE60 G OF STORE--BOUGHT BOUGHT TUMERICTUMERIC SPICE MIXED WITH SPICE MIXED WITH 150 ML DICHLOROMETHANE IN 150 ML DICHLOROMETHANE IN A FLASK A FLASK
REFLUX APPARATUSREFLUX APPARATUS
CONNECTION TO COLD WATERCONNECTION TO COLD WATER
HEATED FOR 3 HOURS IN HOT HEATED FOR 3 HOURS IN HOT WATER BATH (45WATER BATH (45°°C)C)
Water InWater In
Water OutWater Out
CondenserCondenser
Hot Water Hot Water
FlaskFlaskSink OutletSink Outlet
Hot PlateHot Plate
PART 1PART 1: : Thin Layer Chromatography Analysis of CURCUMIN in TurmericThin Layer Chromatography Analysis of CURCUMIN in Turmeric
VACUUM PUMP IS VACUUM PUMP IS CONNECTED TO A FLASK CONNECTED TO A FLASK WITH THE RIGHT SIZE WITH THE RIGHT SIZE TUBINGTUBING
PAPER FILTER IS PLACED PAPER FILTER IS PLACED INSIDE THE FUNNEL INSIDE THE FUNNEL
FUNNEL IS SECURED INTO FUNNEL IS SECURED INTO THE FLASK BY A RUBBER THE FLASK BY A RUBBER STOPPERSTOPPER
Step #2: Suction Filtration Step #2: Suction Filtration
Vacuum PumpVacuum Pump
FlaskFlask
FunnelFunnel
Rubber StopperRubber Stopper
Plastic TubingPlastic Tubing
PART 1PART 1: : Thin Layer Chromatography Analysis of CURCUMIN in TurmericThin Layer Chromatography Analysis of CURCUMIN in Turmeric
DROPPED 3 ML OF SOLVENT (5% METHANOL IN DROPPED 3 ML OF SOLVENT (5% METHANOL IN DICHLOROMETHANE) INTO BEAKER #1 DICHLOROMETHANE) INTO BEAKER #1
SPOTTED A TLC PLATE WITH TURMERIC EXTRACT SPOTTED A TLC PLATE WITH TURMERIC EXTRACT
TLC PLATE IN BEAKER COVERED WITH WATCH GLASSTLC PLATE IN BEAKER COVERED WITH WATCH GLASS
PREPARED 2PREPARED 2NDND BEAKER WITH SAME SOLVENT AS BEAKER WITH SAME SOLVENT AS ABOVE BUT ADDED 3 DROPS OF METHANOL (INCREASE ABOVE BUT ADDED 3 DROPS OF METHANOL (INCREASE POLARITY)POLARITY)
PREPARED PLATE #2 & REPEATEDPREPARED PLATE #2 & REPEATED
THE PLATES ARE TAKEN OUT AS SOON AS THE THE PLATES ARE TAKEN OUT AS SOON AS THE SOLVENT IS 1CM BELOW THE TOP OF THE PLATESOLVENT IS 1CM BELOW THE TOP OF THE PLATE
Step #3: TLC Analysis Step #3: TLC Analysis
Beaker #1Beaker #1
Beaker #2Beaker #2
PART 1PART 1: : Thin Layer Chromatography Analysis of CURCUMIN in TurmericThin Layer Chromatography Analysis of CURCUMIN in Turmeric
Step #3: Results of TLC AnalysisStep #3: Results of TLC Analysis
TLC Plate #1TLC Plate #1 TLC Plate #2TLC Plate #2
ON PLATE #2, SOLVENT ON PLATE #2, SOLVENT TRAVELED 8.10 CM TRAVELED 8.10 CM
THE THE RFRF FOR 1FOR 1STST SPOT SPOT FROM BOTTOM UP IS 0.48FROM BOTTOM UP IS 0.48
THE THE RFRF FOR 2FOR 2NDND SPOT SPOT FROM BOTTOM UP IS 0.67FROM BOTTOM UP IS 0.67
THE THE RFRF FOR 3FOR 3RDRD SPOT SPOT FROM BOTTOM UP IS 0.86FROM BOTTOM UP IS 0.86
THE DARKEST SPOT THE DARKEST SPOT (3(3RDRD SPOT) IS SPOT) IS CURCUMINCURCUMIN..
CURCUMIN TRAVELED CURCUMIN TRAVELED FROM THE BOTTOM UPFROM THE BOTTOM UP
ON PLATE #1, SOLVENT ON PLATE #1, SOLVENT TRAVELED 8.05 CMTRAVELED 8.05 CM
THE THE RFRF FOR 1FOR 1STST SPOT SPOT FROM BOTTOM UP IS 0.30FROM BOTTOM UP IS 0.30
THE THE RFRF FOR 2FOR 2NDND SPOT SPOT FROM BOTTOM UP IS 0.56FROM BOTTOM UP IS 0.56
THE DARKEST SPOT THE DARKEST SPOT (2(2NDND SPOT) IS SPOT) IS CURCUMINCURCUMIN..
RfRf= = distance traveled by spotdistance traveled by spotdistance traveled by solventdistance traveled by solvent
PART 1PART 1: : Thin Layer Chromatography Analysis of CURCUMIN in TurmericThin Layer Chromatography Analysis of CURCUMIN in Turmeric
EVER WONDERED WHAT IS IN YOUR GRAPE SODA?EVER WONDERED WHAT IS IN YOUR GRAPE SODA?
2 SYNTHETIC DYES: Allura Red AC
Brilliant Blue FCF
Allura Red AC can be found in tattoo inks, cosmetics, children's medications, cotton candy.
On June 30, 2010, the Center for Science in the Public Interest (CSPI) called for the FDA to ban Red 40.
Brilliant Blue FCF is a synthetic dye made with aromatic hydrocarbons from petroleum.
Can be found in ice cream, canned processed peas, dairy products, sweets, soaps, shampoos & mouthwash.
It is poorly absorbed from the gastro‐intestinal tract and 95% of the ingested dye can be found in the feces.
PART 2PART 2: : Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
PurposePurpose::
ISOLATION OF THE TWO DYES FOUND IN GRAPE SODA ISOLATION OF THE TWO DYES FOUND IN GRAPE SODA USING A REVERSEUSING A REVERSE--PHASE SEPPHASE SEP--PAKPAK CC--18 18 CARTRIDGE. CARTRIDGE.
CHCH3
Red Dye (FD&C #40), Red Dye (FD&C #40), AlluraAllura RedRed Blue Dye (FD&C #1), Brilliant BlueBlue Dye (FD&C #1), Brilliant Blue
CHCH3
CHCH3
HH3
CC
Cell Cell MenbraneMenbrane
PART 2PART 2: : Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
PREPRE--TREAT THE SEPTREAT THE SEP--PAK CARTRIDGE WITHPAK CARTRIDGE WITH
10.0 ML OF 70% ISOPROPYL ALCOHOL 10.0 ML OF 70% ISOPROPYL ALCOHOL SOLUTION SOLUTION
10.0 ML OF WATER.10.0 ML OF WATER.
FORCE 5.0 ML OF GRAPE SODA THE SEPFORCE 5.0 ML OF GRAPE SODA THE SEP--PAK PAK USING A SYRINGE. USING A SYRINGE.
COLLECT THE 1COLLECT THE 1STST FRACTION (FRACTION (ELUENTELUENT).).
Step #1: Separation of the Red Dye (FD&C #40) Step #1: Separation of the Red Dye (FD&C #40)
PART 3PART 3::Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
FORCE 5.0 ML 5% ISOPROPYL FORCE 5.0 ML 5% ISOPROPYL ALCOHOL SOLUTION THROUGH ALCOHOL SOLUTION THROUGH THE SEPTHE SEP--PAK. PAK.
COLLECT THE 2COLLECT THE 2NDND FRACTION FRACTION
REPEAT 3 X. THIS YIELDS REPEAT 3 X. THIS YIELDS FRACTIONS 3 THROUGH 5.FRACTIONS 3 THROUGH 5.
PART 3PART 3: : Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
FORCE 2.5 ML 5% ISOPROPYL FORCE 2.5 ML 5% ISOPROPYL ALCOHOL SOLUTION THROUGH ALCOHOL SOLUTION THROUGH THE SEPTHE SEP--PAK. PAK.
COLLECT THE 6COLLECT THE 6THTH FRACTION FRACTION
REPEAT 4 X. THIS YIELDS REPEAT 4 X. THIS YIELDS FRACTIONS 7 THROUGH 10.FRACTIONS 7 THROUGH 10.
PART 3PART 3: : Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
Step #2: Separation of the Blue Dye (FD&C #1) Step #2: Separation of the Blue Dye (FD&C #1)
FORCE 5.0 ML 20% ISOPROPYL FORCE 5.0 ML 20% ISOPROPYL ALCOHOL SOLUTION THROUGH ALCOHOL SOLUTION THROUGH THE SEPTHE SEP--PAK. PAK.
COLLECT THE 11COLLECT THE 11THTH FRACTION FRACTION
REPEAT 3 X. THIS YIELDS REPEAT 3 X. THIS YIELDS FRACTIONS 12 THROUGH 14.FRACTIONS 12 THROUGH 14.
PART 3PART 3: : Separation and Analysis of Dyes in Grape SodaSeparation and Analysis of Dyes in Grape Soda
SAMPLESSAMPLES’’ OPTICAL OPTICAL DENSITIES ARE READ WITH DENSITIES ARE READ WITH A SPECTROPHOTOMETER. A SPECTROPHOTOMETER.
AMOUNT OF DYE IN ONE CAN OF GRAPE SODA:AMOUNT OF DYE IN ONE CAN OF GRAPE SODA:
•• 2.59 MG OF RED DYE2.59 MG OF RED DYE•• 0.36 MG OF BLUE DYE 0.36 MG OF BLUE DYE
SOURCES_____________________________________
http://en.wikipedia.org/wiki/Stearic_Acidhttp://en.wikipedia.org/wiki/Stearic_Acidhttp://qwickstep.com/search/sodiumhttp://qwickstep.com/search/sodium--chloridechloride--inin--water.htmlwater.htmlhttp://www.3dchem.com/molecules.asp?ID=176http://www.3dchem.com/molecules.asp?ID=176http://http://www.btinternet.com/~martin.chaplin/molecule.htmlwww.btinternet.com/~martin.chaplin/molecule.htmlhttp://www.worldofmolecules.com/3D/turmeric_3d.htmhttp://www.worldofmolecules.com/3D/turmeric_3d.htmhttp://upload.wikimedia.org/wikipedia/commons/6/67/Turmericroot.http://upload.wikimedia.org/wikipedia/commons/6/67/Turmericroot.jpgjpghttp://http://en.wikipedia.org/wiki/Allura_Red_ACen.wikipedia.org/wiki/Allura_Red_AChttp://http://en.wikipedia.org/wiki/Brilliant_Blue_FCFen.wikipedia.org/wiki/Brilliant_Blue_FCFhttp://en.wikipedia.org/wiki/Allura_Red_AChttp://en.wikipedia.org/wiki/Brilliant_Blue_FCFhttp://http://www.northallegheny.orgwww.northallegheny.orghttp://http://nih.govnih.gov//http://drvikram.com/herbalhttp://drvikram.com/herbal‐‐products.htmproducts.htmhttp://www.madhealth.net/curcuminhttp://www.madhealth.net/curcumin‐‐preventsprevents‐‐cancercancer‐‐heartheart‐‐diseasedisease‐‐alzheimer%E2%80%99salzheimer%E2%80%99s‐‐diseasedisease‐‐andand‐‐obesity/obesity/http://www.themoneytimes.com/files/cancer%20cells.jpghttp://www.themoneytimes.com/files/cancer%20cells.jpg
THANK YOU TO PROFESSOR KIMBERLY STIEGLITZ
_______________________
PRESENTED BY HELENE GAUDETTE
FOR SCI 124 PRINCIPLES OF CHEMISTRY II
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