sci 124 new lab chromatographic separation methods

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


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


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


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


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


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.



COLLECT THE 6COLLECT THE 6THTH FRACTION FRACTION



Step #2: Separation of the Blue Dye (FD&C #1) Step #2: Separation of the Blue Dye (FD&C #1)


COLLECT THE 11COLLECT THE 11THTH FRACTION FRACTION



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

sci 124 new lab chromatographic separation methods

Documents

analysis of dyes

grape soda step

grape soda force

grape sodapurpose

grape sodausing

grape sodastep

red dye fdc

turmeric extract tlc