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Sci 124 New Lab Chromatographic Separation Methods

May 26, 2015

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  • 1. CHROMATOGRAPHIC SEPARATION METHODSSEPARATION BY THIN-LAYER CHROMATOGRAPHY (TLC)& REVERSE-PHASE

2. CURCUMIN, ANYONE ? 1,529,560NewCancerCasesin2010569,490CancerDeathsin2010CurcuminPreventsCancer,HeartDisease,AlzheimersDiseaseandObesity.CancerCell 3. PART 1: Thin Layer Chromatography Analysis of CURCUMIN in TurmericPurpose: ISOLATION OF THE YELLOW PIGMENT CURCUMIN FOUND IN THE SPICE TUMERIC BY THIN-LAYER CHROMATOGRAPHY (TLC) METHOD & USING A POLAR ORGANIC SOLVENT.CurcuminTurmericRootTUMERIC COMES FROM THE CURCUMA LONGAPLANT (GINGER-LIKE) FOUND IN ASIA.CellMenbrane:Phospholipid Bilayer 4. PART 1: Thin Layer Chromatography Analysis of CURCUMIN in Turmeric Step #1: Heating Reflux60 G OF STORE-BOUGHTTUMERIC SPICE MIXED WITH150 ML DICHLOROMETHANE IN Water OutA FLASKCondenser REFLUX APPARATUS Water InFlask CONNECTION TO COLD WATERSink OutletHot Water HEATED FOR 3 HOURS IN HOTHot PlateWATER BATH (45C) 5. PART 1: Thin Layer Chromatography Analysis of CURCUMIN in Turmeric Step #2: Suction Filtration FunnelVACUUM PUMP IS Rubber StopperCONNECTED TO A FLASK Plastic TubingWITH THE RIGHT SIZETUBINGPAPER FILTER IS PLACEDINSIDE THE FUNNELFlask Vacuum Pump FUNNEL IS SECURED INTOTHE FLASK BY A RUBBERSTOPPER 6. PART 1: Thin Layer Chromatography Analysis of CURCUMIN in TurmericStep #3: TLC Analysis DROPPED 3 ML OF SOLVENT (5% METHANOL INDICHLOROMETHANE) INTO BEAKER #1 SPOTTED A TLC PLATE WITH TURMERIC EXTRACT TLC PLATE IN BEAKER COVERED WITH WATCH GLASSBeaker #1 PREPARED 2ND BEAKER WITH SAME SOLVENT ASABOVE BUT ADDED 3 DROPS OF METHANOL (INCREASEPOLARITY) PREPARED PLATE #2 & REPEATED THE PLATES ARE TAKEN OUT AS SOON AS THESOLVENT IS 1CM BELOW THE TOP OF THE PLATEBeaker #2 7. PART 1: Thin Layer Chromatography Analysis of CURCUMIN in Turmeric Step #3: Results of TLC Analysis CURCUMIN TRAVELEDON PLATE #2, SOLVENTFROM THE BOTTOM UP TRAVELED 8.10 CM ON PLATE #1, SOLVENT THE RF FOR 1ST SPOTTRAVELED 8.05 CM FROM BOTTOM UP IS 0.48 THE RF FOR 1ST SPOTTHE RF FOR 2ND SPOTFROM BOTTOM UP IS 0.30 FROM BOTTOM UP IS 0.67 THE RF FOR 2ND SPOTTHE RF FOR 3RD SPOTFROM BOTTOM UP IS 0.56 FROM BOTTOM UP IS 0.86THE DARKEST SPOT THE DARKEST SPOT(2ND SPOT) IS CURCUMIN.(3RD SPOT) IS CURCUMIN.Rf=distancetraveledbyspotdistancetraveledbysolventTLC Plate #1TLC Plate #2 8. EVER WONDERED WHAT IS IN YOUR GRAPE SODA? 2 SYNTHETIC DYES:Allura RedAC BrilliantBlueFCFAllura RedACcanbefoundin tattooinks,cosmetics,childrensmedications,cottoncandy.OnJune30,2010,theCenterforScienceinthePublicInterest (CSPI)calledfortheFDAtobanRed40.BrilliantBlueFCFisasyntheticdyemadewitharomatichydrocarbonsfrompetroleum.Canbefoundinicecream,cannedprocessedpeas,dairyproducts,sweets,soaps,shampoos&mouthwash.Itispoorlyabsorbedfromthegastrointestinaltractand95%oftheingesteddyecanbefoundinthefeces. 9. PART 2: Separation and Analysis of Dyes in Grape SodaPurpose:ISOLATION OF THE TWO DYES FOUND IN GRAPE SODAUSING A REVERSE-PHASE SEP-PAK C-18CARTRIDGE. CH 3CellMenbrane CH 3CH H C3 3Red Dye (FD&C #40), Allura Red Blue Dye (FD&C #1), Brilliant Blue 10. PART 2: Separation and Analysis of Dyes in Grape SodaStep #1: Separation of the Red Dye (FD&C #40)PRE-TREAT THE SEP-PAK CARTRIDGE WITH 10.0 ML OF 70% ISOPROPYL ALCOHOLSOLUTION10.0 ML OF WATER.FORCE 5.0 ML OF GRAPE SODA THE SEP-PAKUSING A SYRINGE. COLLECT THE 1ST FRACTION (ELUENT). 11. PART 3:Separation and Analysis of Dyes in Grape Soda FORCE 5.0 ML 5% ISOPROPYLALCOHOL SOLUTION THROUGHTHE SEP-PAK. COLLECT THE 2ND FRACTION REPEAT 3 X. THIS YIELDSFRACTIONS 3 THROUGH 5. 12. PART 3: Separation and Analysis of Dyes in Grape Soda FORCE 2.5 ML 5% ISOPROPYLALCOHOL SOLUTION THROUGHTHE SEP-PAK. COLLECT THE 6TH FRACTION REPEAT 4 X. THIS YIELDSFRACTIONS 7 THROUGH 10. 13. PART 3: Separation and Analysis of Dyes in Grape Soda Step #2: Separation of the Blue Dye (FD&C #1) FORCE 5.0 ML 20% ISOPROPYLALCOHOL SOLUTION THROUGHTHE SEP-PAK. COLLECT THE 11TH FRACTION REPEAT 3 X. THIS YIELDSFRACTIONS 12 THROUGH 14. 14. PART 3: Separation and Analysis of Dyes in Grape Soda SAMPLES OPTICAL DENSITIES ARE READ WITH A SPECTROPHOTOMETER.AMOUNT OF DYE IN ONE CAN OF GRAPE SODA: 2.59 MG OF RED DYE 0.36 MG OF BLUE DYE 15. SOURCES _____________________________________http://en.wikipedia.org/wiki/Stearic_Acidhttp://qwickstep.com/search/sodium-chloride-in-water.htmlhttp://www.3dchem.com/molecules.asp?ID=176http://www.btinternet.com/~martin.chaplin/molecule.htmlhttp://www.worldofmolecules.com/3D/turmeric_3d.htmhttp://upload.wikimedia.org/wikipedia/commons/6/67/Turmericroot.jpghttp://en.wikipedia.org/wiki/Allura_Red_AChttp://en.wikipedia.org/wiki/Brilliant_Blue_FCFhttp://en.wikipedia.org/wiki/Allura_Red_AChttp://en.wikipedia.org/wiki/Brilliant_Blue_FCFhttp://www.northallegheny.orghttp://nih.gov/http://drvikram.com/herbalproducts.htmhttp://www.madhealth.net/curcuminpreventscancerheartdiseasealzheimer%E2%80%99sdiseaseandobesity/http://www.themoneytimes.com/files/cancer%20cells.jpg 16. THANK YOU TO PROFESSORKIMBERLY STIEGLITZ_______________________PRESENTED BYHELENE GAUDETTE FORSCI 124 PRINCIPLES OF CHEMISTRY II

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