ADVANCE CHEMISTRY

ISOLATION
OF
CAFFEINE
FROM TEA
SOLUTION
WEDNESDAY, SEPTEMBER 19, 2001

SUPARADA SUWANLERTLUN  5961084

THAYA NGAMPRAVATDEE  5961075
SUPITCHAYANUT SITTISUPAROGE  5961045
NANNAPHAT YONGPARPHAT  5961003
 ABSTRACT

          THE PURPOSE OF THIS LAB IS TO EXTRACT THE PURE

CAFFEINE FROM TEA SAMPLE AND TO KNOW MORE SOME
METHODS THAT CAN USE TO IDENTIFY THE SUBSTANCE. WE
USE FOUR PROCEDURES INCLUDING EXTRACTION,
FILTRATION, TLC (BY COMPARING WITH THE PURE ONE THAT
WE ALREADY KNOW), AND MELTING POINT TO IDENTIFY THAT
OUR EXTRACTION IS PURE OR NOT. THERE WAS AN ERROR
DURING THE EXPERIMENT AS WE DO NOT KNOW THE EXACT
POINT THAT IT IS START TO MELT. HOWEVER, THE RESULT OF
THIS EXPERIMENT IS OUR EXTRACTION IS NOT PURE, IT CAN
BE NOTICED BECAUSE OUR SUBSTANCE BEGINS TO MELT
BEFORE THE PURE CAFFEINE SAMPLE, SO WE CAN ASSUME
THAT IT IS CONTAMINATED.
 INTRODUCTION

          PEOPLE MIGHT NOT KNOWN THAT MOST BEVERAGES

TAKEN IN PRESENT CONTAINS CAFFEINE INSIDE. THIS TYPE
OF COMPOUND CAN BE FOUND IN THINGS PEOPLE INGESTED
IN THEIR EVERYDAY LIFE SUCH AS COFFEE, TEA, COCOA,
MILK CHOCOLATE, BAKING CHOCOLATE, COCA-COLA,
EXCEDRIN,AND OR EVEN IN NO-DOZ. CAFFEINE
(C8H10N4O2) IS A METHYLXANTHINE CONSIDERED AS A
MEMBER OF THE CLASS CALLED ALKALOIDS IN ORGANIC
COMPOUNDS (PUBCHEM.NCBI.NLM.NIH.GOV, 2018) [1].
NATURALLY, CONTAINING BASIC COMPOUNDS SUCH AS
NITROGEN THAT CAN BE FOUND IN PLANTS. THE TASTE OF
CAFFEINE IS COMMONLY BITTER. CAFFEINE IS MOSTLY USED
AS A PHARMACOLOGICAL AGENT AFFECTED IN HUMAN’S
NERVOUS SYSTEM HELPING IN INCREASE IN ALERTNESS AND
AGITATION. SOME OF THESE COMPOUNDS INCLUDED:
NICOTINE, COCAINE, MORPHINE, AND STRYCHNINE. A
DIAGRAM OF CAFFEINE’S STRUCTURE IS GIVEN BELOW. 
          CAFFEINE IS HELPFUL IN RELAXING SMOOTH
MUSCLES, STIMULATING CARDIAC MUSCLES AND
DIURESIS AND ALSO TREATING TYPE OF HEADACHE.
HOWEVER, IT IS PHYSICALLY ADDICTIVE. PEOPLE WITH
CAFFEINE ADDICTION AND TEND TO STOP TAKING IT MAY
POSSIBLY EXPERIENCES INSOMNIA, HEADACHE, AND
NAUSEA. 
          HOWEVER, IN THIS LAB, WE WILL MAINLY USE THE
EXTRACTION METHOD. THIS METHOD WILL SEPARATE THE
SUBSTANCES WHEN THEY ARE MIXED TOGETHER IN
IMMISCIBLE PHRASE SUCH AS OIL AND WATER IN THIS LAB
(CHEMICOOL.COM, 2018) [2]. IN ADDITION, THE SUBSTANCE
THAT IS LESS SOLUBLE WILL MOVE TO THE SUBSTANCE THAT
IS MORE SOLUBLE, SO WE CAN SEE THE SEPARATION OF TWO
LAYERS IN THIS METHOD AND USE THE STOPCOCK TO
CONTROL THE VOLUME THAT WE WANT. HOWEVER, THE
SUBSTANCE THAT HAS MORE DENSITY WILL PRESENT IN
LOWER PART AND THE SUBSTANCE THAT HAS LESS DENSITY
WILL PRESENT IN THE UPPER PART. FOR EXAMPLE, WHEN
EXTRACT BETWEEN WATER AND TRICHLOROMETHANE,
TRICHLOROMETHANE IS MORE DENSE, SO IT WILL PRESENT IN
THE LOWER PART IN THE SEPARATORY FUNNEL AND WATER IS
LESS DENSE, SO IT WILL PRESENT IN THE UPPER PART IN THE
SEPARATORY FUNNEL.
          AFTER THAT, WE WILL USE TLC (THIN LAYER
CHROMATOGRAPHY), IT IS THE METHOD THAT USED TO
SEPARATE MIXTURES OF SUBSTANCES INTO THEIR
COMPONENTS. ALL FORMS OF CHROMATOGRAPHY WORK ON
THE SAME PRINCIPLE (CHEMGUIDE.CO.UK, 2018) [3]. IN
THIS METHOD, IT HAS 2 PHASES WHICH ARE STATIONARY
PHASE AND MOBILE PHASE. IN ADDITION, STATIONARY
PHASE IS THE PHASE THAT SUPPORT THE MOBILE PHASE.
THE MOBILE PHASE WILL FLOW THROUGH THE STATIONARY
PHASE AND CARRY THE COMPONENTS AND MIXTURES WITH
IT. ALSO, DIFFERENT COMPONENTS WILL FLOW IN THE
DIFFERENT RATE (CHEMISTRY LIBRETEXTS, 2018) [4].
PURPOSE
TO EXTRACT CAFFEINE FROM TEA SOLUTION AND TO LEARN
MORE ABOUT THE METHODS THAT ARE USED IN THIS
EXPERIMENT SUCH AS EXTRACTION, FILTRATION, TLC, AND
MELTING POINT. 

MATERIALS
TEA BAGS
50 ML ERLENMEYER FLASK
125 ML SEPARATORY FUNNEL
RING STAND
DICHLOROMETHANE
GLASS ROD
CAPILLARY TUBE
TLC PLATE
ROTAEVAPORATORY
VACUUM
TLC CAMBER
METHOD
1. TRANSFER THE TEA EXTRACT FROM THE 50 ML
ERLENMEYER FLASK TO A 125 ML SEPARATORY FUNNEL
THAT IS SUPPORTED BY A RING ON A RING STAND.
2. ADD 20 MILLILITERS OF DICHLOROMETHANE TO THE
SEPARATORY FUNNEL. STOPPER THE FUNNEL AND GRAB THE
NECK OF THE FUNNEL WITH ONE HAND, HOLDING THE
STOPPER INTO THE NECK OF THE FUNNEL. WITH YOUR
OTHER HAND GRASP THE STOPCOCK IN SUCH A WAY THAT
YOU CAN TURN THE PLUG IN THE BARREL TO OPEN AND
CLOSE THE STOPCOCK. WHILE HOLDING THE STOPPER
TIGHTLY INTO THE NECK OF THE FUNNEL, INVERT THE
FUNNEL SO THE LIQUID NO LONGER IS IN CONTACT WITH
THE STOPCOCK. POINTING THE STEM OF THE FUNNEL AWAY
FROM EVERYBODY, OPEN THE STOPCOCK TO RELEASE ANY
PRESSURE THAT MAY HAVE BUILT UP INSIDE THE FUNNEL.
CLOSE THE STOPCOCK AND AGITATE THE MATERIAL IN THE
FUNNEL WITHOUT SHAKING IT VIGOROUSLY – YOU WANT
THE CONTENTS TO MIX, BUT YOU DO NOT WANT TO
GENERATE AN EMULSION. POINT THE STEM OF THE FUNNEL
UPWARD, AND MAKING SURE NO LIQUID IS IN CONTACT
WITH THE STOPCOCK OPEN IT TO AGAIN RELIEVE ANY
PRESSURE BUILD-UP. CLOSE THE STOPCOCK. AGITATE
AGAIN AS BEFORE AND RECLAMP THE SEPARATORY FUNNEL
TO THE RING STAND.
3. ALLOW THE CONTENTS OF THE SEPARATORY FUNNEL
TO SETTLE. THERE SHOULD BE TWO DISTINCT MOSTLY
CLEAR LAYERS. IF THERE IS AN EMULSION (CLOUDY)
LAYER BETWEEN TWO CLEAR LAYERS IT IS SOMETIMES
POSSIBLE TO BREAK THE EMULSION BY SWIRLING THE
CONTENTS OF THE FUNNEL OR STIRRING THE CONTENTS
USING A GLASS ROD. 
4. CAREFULLY DRAIN THE LOWER (DICHLOROMETHANE)
LAYER INTO A 25 ML ERLENMEYER FLASK. TRY TO NOT
INCLUDE ANY OF THE AQUEOUS (UPPER) LAYER. IF THERE
IS A LOT OF EMULSION, INCLUDE IT IN THE ERLENMEYER
FLASK.
5. REPEAT STEPS 2 THROUGH 4 USING A SECOND 20 ML
PORTION OF DICHLOROMETHANE.
6. COMBINE ORGANIC LAYERS FROM STEP 2 - 4, THEN
WASH IT WITH 20 ML OF 6M NAOH
7. CAREFULLY DRAIN THE LOWER (DICHLOROMETHANE)
LAYER INTO A 25 ML ERLENMEYER FLASK. TRY TO NOT
INCLUDE ANY OF THE AQUEOUS (UPPER) LAYER.
8. WASH THE ORGANIC LAYER FROM STEP 7 WITH 20 ML
OF H2O, THEN ROTOVAP THE SUBSTANCE TO DRYNESS
9. RINSE THE RESIDUE, AND WEIGHT THE DRIED
SUBSTANCE
10. PREPARE TLC PLATE BY CUTTING IT INTO
RECTANGULAR SHAPE, DRAW FINISH LINE (1 CM FROM
THE UPPER EDGE) AND START LINE (1.5 CM FROM THE
BOTTOM EDGE). ON START LINE, ASSIGN TWO DOT WHICH
ARE FOR THE EXTRACTION WE MADE AND THE PURE
CAFFEINE SAMPLE.  
11. DISSOLVE SOME PART OF DRIED CAFFEINE
EXTRACTION WITH DISTILLED WATER, AND USE SHARP-
END OF A CAPILLARY TUBE TO TRANSFER THE SOLUTION
FROM PETRI DISH, DROP SOME OF IT ON PREPARED TLC
PLATE (ON THE DOT THAT IS PROVIDED FOR EXTRACTION).
12. DISSOLVE THE PURE CAFFEINE, THEN USE SHARP-END
OF A CAPILLARY TUBE TO TRANSFER THE SOLUTION FROM
PETRI DISH, DROP SOME OF IT ON PREPARED TLC PLATE
(ON THE DOT THAT IS PROVIDED FOR PURE CAFFEINE
SAMPLE).
13. PLACE THE TLC PLATE IN TO A GLASS THAT FILLED
DICHLOROMETHANE (ABOUT 1 CM HIGH)
14. OBSERVE THE RESULT
15. AFTER GETTING RESULTS FROM TLC TEST. CAREFULLY
PUT THE CRUDES AND PURE CAFFEINE SUBSTANCES IN
SEPARATE CAPILLARY TUBES. SO, WE WILL GET TWO
CAPILLARY TUBES TO RUN IN THE NEXT MELTING POINT
TEST.
16. PUT THE THERMOMETER INSIDE THE MELTING POINT
TO CHECK FOR TEMPERATURE RANGE ALONG THE TEST.
17. THEN PUT BOTH OF CAPILLARY TUBES CONTAINING
OUR CRUDES AND PURE SUBSTANCES BESIDE TOGETHER
TO COMPARE THEIR MELTING POINT.
18. WAIT FOR THE TEMPERATURE TO HIT AROUND 200°C
THEN LOOKS THROUGH THE MELTING POINT MACHINE TO
CHECK FOR CHANGED.
19. RECORD THE RANGE OF TEMPERATURE IF ONE OF THE
SUBSTANCES MELT.
RESULT
          AT FIRST, WE EXTRACTED CAFFEINE FROM TEA BY
USING DICHLOROMETHANE AND GET OUR CAFFEINE
CRUDES. THEN AFTER GETTING CAFFEINE CRUDES FROM
EXTRACTION, WE TEST THE PURITY OF OUR CRUDES BY
USING TLC PAPER AND CHAMBER. THE RESULTS COME
OUT THAT WE GOT CAFFEINE FROM THE TEA EXTRACTION
BECAUSE THE SPOT IS IN THE SAME POSITION AS OUR
REFERENCE OR PURE CAFFEINE SUBSTANCE. HOWEVER,
OUR CRUDES WERE NOT PURE SUBSTANCES BECAUSE IT
ALSO CONTAIN ANOTHER SPOT ABOVE WHICH MEAN THAT
THERE ARE CONTAMINATED IN CRUDES. NEXT, WE PUT
OUR CRUDES INTO CAPILLARY TUBE AND ANOTHER PURE
CAFFEINE SUBSTANCE INTO ANOTHER TUBE AND THEN
RUN THEM IN A MELTING POINT MACHINE FOR TEST. FROM
DETERMINING MELTING POINT OF OUR CAFFEINE CRUDE
AND PURE SUBSTANCES OF CAFFEINE, OUR GROUP HAVE
FOUND OUT THAT OUR CRUDES WERE THE FIRST ONE TO
DISSOLVE AND TRANSFORM INTO LIQUID SUBSTANCES
AND THEN FOLLOWING BY MELTING OF PURE SUBSTANCES
OF CAFFEINE. HOWEVER, WE HAVE AN ERROR ON DOING
THIS STEP BECAUSE WE CANNOT CATCH UP THE RANGE
OF CELSIUS THAT OUR SUBSTANCES MELTED UP.
ACCORDING TO REPORTED IN LITERATURE, THE MELTING
POINT OF CAFFEINE SHOULD BE 238°C. SO, OUR RANGE
OF TEMPERATURE SHOULD BE AROUND 200°C BUT NOT
EXCEED 260°C.
DISCUSSION
          BASED ON THEORY, WE ALREADY KNOW THE
PROPERTIES OF SUBSTANCE CONSISTING OF MELTING
POINT AND THIN LAYER CHROMATOGRAPHY. OUR
SUBSTANCE HAS ALMOST THE EXACT SAME HEIGHT AS
THE PURE ONE, BUT THE PURE ONE IS A BIT HIGHER THAN
OUR SUBSTANCE. THEREFORE, OUR SUBSTANCE MIGHT BE
CONTAMINATED SINCE THE OTHER SUBSTANCES THAT
WERE MIXED WITH EXTRACTED CAFFEINE PROBABLY HAVE
STRONGER ATTRACTION TO PULL CAFFEINE DOWN, SO IT
DID NOT GO AS HIGH AS THE ORIGINAL ONE. REGARDING
MELTING POINT, OUR CAFFEINE MELTED BEFORE THE
PURE CAFFEINE, HENCE IT IS OBVIOUS TO ASSUME THAT
OUR CAFFEINE IS NOT PURE AND CONTAMINATED.

1. EXPLAIN WHY EMULSION CAN BE OBSERVED DURING

LIQUID-LIQUID EXTRACTION? HOW TO AVOID IT? IF IT
OCCURS, WHAT IS SIMPLE METHODS TO DESTROY IT?
BOTH OF THE SUBSTANCES INCLUDING CAFFEINE AND
DICHLOROMETHANE, ARE POLAR, HENCE EMULSION CAN
BE HAPPENED. THE WAY TO AVOID EMULSION IS NOT
SWIRL THE SOLUTION TOO MUCH, SO THAT THE
SUBSTANCES WOULD BE MIXED, BUT IF THERE IS
EMULSION, IT CAN SIMPLY REMOVE USING GLASS ROD.
2. WHY DO WE NEED DRYING AGENT (NA2SO4) AFTER
FINISHING LIQUID-LIQUID EXTRACTION AND BEFORE
EVAPORATION PROCESS?
BECAUSE NA2SO4 CAN ABSORB WATER, SO OUR
EXTRACTION WILL LOSS AMOUNT OF EXCESS WATER AND
REMAIN PURE CAFFEINE.

3. HOW DO YOU IDENTIFY THAT YOU GET CAFFEINE FROM

TEA LEAVES? IS IT PURE?
ACCORDING TO THE EXPERIMENT, THE PURE CAFFEINE IS
PROVIDED TO CLARIFY THE EXPECTED RESULTS FOR THE
TEST CONSISTING OF MELTING POINT AND
CHROMATOGRAPHY, SO THE EXTRACTION CAN BE
COMPARED TO IT. THE RESULT IS THAT THE EXTRACTION
IS NOT PURE. AS COMPARING TO THE INITIAL RESULT,
THE MELTING POINT AND RESULT OF CHROMATOGRAPHY
ARE NOT THE SAME.

4. WHAT IS THE RF VALUE OF CAFFEINE? 

5. WHAT ARE ERRORS DURING THE EXPERIMENT AND HOW
TO AVOID THEM OR IMPROVE YOUR TECHNIQUE?
THE ERRORS THAT APPEARED WERE MISUNDERSTANDING
THE METHOD AND COULD NOT GET THE EXACT
TEMPERATURE OF MELTING POINT. ALTHOUGH THE
EXTRACTION WAS NOT CONTAMINATED, MIXING WRONG
SUBSTANCES COULD BE VERY DANGEROUS. THEREFORE,
IN THE NEXT EXPERIMENT, THE EXPERIMENTERS SHOULD
SPEND MORE TIME ON PROCEDURE TO MAKE SURE THAT
THEY ARE CLEARLY UNDERSTAND HOW TO PERFORM THE
EXPERIMENT.  OTHER THAN THAT, WE SHOULD BE MORE
CAREFUL FOR OBSERVING TEMPERATURE OR EVEN OTHER
REACTION THAT COULD BE REPORTED. SO, THE RESULT
OF THE EXPERIMENT IS MORE ACCURATE AND RELIABLE. 
  
6. DETERMINE THE STRONGEST INTERMOLECULAR FORCES
PRESENT IN CAFFEINE.
THE STRONGEST INTERMOLECULAR FORCE WHICH
PRESENTS IN CAFFEINE IS DIPOLE-DIPOLE FORCE
BECAUSE EACH MOLECULE OF CAFFEINE HAS A DIPOLE-
DIPOLE FORCE WITH DIFFERENT EN, SO THEY WILL HAVE
DIPOLE-DIPOLE FORCE WITH EACH OTHER WHEN THEY ARE
BOUND.
 CONCLUSION
          AFTER DOING THIS LAB, THE EXTRACTION OF
CAFFEINE FROM TEA SOLUTION, WE HAVE GOT CRUDES OF
CAFFEINE AS A PRODUCT. ALSO, AFTER RUNNING TLC
TEST WE KNOWN THAT OUR CRUDES ARE CAFFEINE BUT
ALSO MIXTURE AND CONTAMINATED WITH OTHER
SUBSTANCES. LASTLY, WE RUN OUR CRUDES IN MELTING
POINT MACHINE AND GOT A RESULT THAT OUR CRUDES
MELT UP FIRST SO ITS PURITY IS NOT PURE AS PURE
SUBSTANCE WHICH CONFIRMED THE POINT THAT OUR
CRUDES ARE CONTAMINATED.
 REFERENCES (APA)
[1]  PUBCHEM.NCBI.NLM.NIH.GOV. (2018). CAFFEINE.
[ONLINE] AVAILABLE AT:
HTTPS://PUBCHEM.NCBI.NLM.NIH.GOV/COMPOUND/CAFFE
INE#SECTION=TOP [ACCESSED 13 SEP. 2018].

[2] CHEMICOOL.COM. (2018). DEFINITION OF

EXTRACTION - CHEMISTRY DICTIONARY. [ONLINE]
AVAILABLE AT:
HTTPS://WWW.CHEMICOOL.COM/DEFINITION/EXTRACTION
.HTML [ACCESSED 13 SEP. 2018].

[3] CHEMGUIDE.CO.UK. (2018). THIN LAYER

CHROMATOGRAPHY. [ONLINE] AVAILABLE AT:
HTTPS://WWW.CHEMGUIDE.CO.UK/ANALYSIS/CHROMATOG
RAPHY/THINLAYER.HTML [ACCESSED 17 SEP. 2018].

[4] CHEMISTRY LIBRETEXTS. (2018). THIN LAYER

CHROMATOGRAPHY. [ONLINE] AVAILABLE AT:
HTTPS://CHEM.LIBRETEXTS.ORG/DEMOS%2C_TECHNIQUE
S%2C_AND_EXPERIMENTS/GENERAL_LAB_TECHNIQUES/T
HIN_LAYER_CHROMATOGRAPHY [ACCESSED 17 SEP.
2018].