CHM141-L ORGANIC CHEMISTRY 1

POST-LABORATORY REPORT (4)

ISOLATION OF CAFFEINE FROM DRIED TEA LEAVES
Jean Paul Manapat , Lian Justin Salundaga, John Rex Tadena
BS CHEMICAL ENGINEERING MAPUA INSTIUTE OF TECHONOLOGY AT LAGUNA

RESULTS AND DISCUSSION

Alkaloids are naturally occurring organic compounds from certain plants which are
distinguishable with the basic nitrogen usually present in a heterocyclic ring like pyrrole, pyridine,
pyrrolidine, quinolone, or isoquinoline. Alkaloids can also be found in an aliphatic side chain. Since
it is an ammonia derivative, most of alkaloids occurs in secondary or tertiary amines. These organic
compounds have wide applications within the realm of pharmacology and medicine. It is found in
different parts of plants as salts of inorganic or organic plant acids in complex mixture with water-
insoluble compounds like gums, tannins, lipids, proteins and mineral salts.

(A) (B)

Caffeine is one example of alkaloid that is widely used as a psychostimulant drug and is
taken in to consumption in the form of coffee. This highly consumed compound serves as a
protection for plants by serving as a natural pesticide that is toxic for some insects and pests. The
structure of caffeine and adenosine shows similar characteristics on their solubility on water. This
enables them to bind at the adenosine receptors present in the brain that increases nerve activity ,
thus making people increase their level of alertness and become more euphoric, increased their
endurance, and state of productivity. Its negative effects is adherent for causing addiction ,
insomnia and aggravates heart conditions. This compound can be found on 28 plants and on this
experiment will focus on the extraction of caffeine from dried tea leaves (Figure 4.1B).that comes
from tea plant or Camelia sinensis . There are varieties of tea leaves that have different amounts of
caffeine content (1) First is the black tea that is fermented that approximately contains 40-70 mg of
caffeine per 8 oz . (2) The green teathat has lesser amounts than black tea and contains 35-45 mg
of caffeine and is unfermented . (3) White tea having the lowest amount of caffeine with 37-55 mg
of caffeine (4) Ooling tea that has a middle range from black tea with 37-55 mg per 8 oz and is
partially fermented and lastly the (5) herbal tea that almost contains no caffeine.

However, caffeine does not subsist in tea leaves individually, since the
composition of tea leaves comprises mostly of cellulose fibers, pigments, glucose, chlorophylls and
other soluble phenolic compounds that contains mostly of hydroxide bonds that is directly attached
to it’s aromatic ring also known as tannins (Rebecca et.al 2014) . In order to extract caffeine from
the vast composition of tea leaves one must utilized several process such as distillation and solvent
extraction. Solvent extraction is a method to separate the desired compound from the rest of the
mixture by taking advantage of the solubility of the desired component to different types of
extracting solvent, a polar and non-polar solvent. On the experiment, there are two types of
extraction that is utilized, first is the solid-liquid extraction that is used or separating the water
soluble components from the insoluble components of tea leaves , observations on the soild-liquid
extraction is summarized in Table 4.1.

Procedure Observation
Mixing of tea leaves and sodium carbonate Dark liquid solution
Addition of distilled water Opaque dark yellow solution
Heating the mixture Darker shade liquid solution
Decantation to 250 mL E-flask Dark-colored solution
Cooling the room temperature Dark-colored solution
Extraction with dichloromethane Formation of bubbles, separation of two
solutions.
Cloudy solution with impurities, brown spots
Addition with sodium sulfate Clear solution
Table 4.1. Extraction of Caffeine from Dried Tea Leaves using Solid-Liquid and Liquid-Liquid Extraction.
 As aforementioned, since tannin and caffeine are soluble in water at high temperature they
can be separated through solid liquid extraction. Preliminary to the extraction , a heterogenous mixture
of 10 g of dried tea leaves with sodium carbonate and boiling chips was made with 87.5 mL of distilled
water resulting in a dark liquid solution. The solubility of caffeine in water is approximately 22 mg per ml
at 25 degrees Celsius however since the solubility and temperature implies a directly proportional
relationship this solubility will increase as water is heated to 300 degrees Celsius resulting in a darker
shade of liquid solution since more desired quantities are dissolved. The heating will leave the solid
insoluble compounds and can be decant to separate it from the supernatant as preparation for the
liquid-liquid extraction. On this process sodium carbonate is added to the boiled mixture of tea leaves
and distilled water since this will act as a base and convert the tannins into water-soluble salts with a
chemical reaction of
ArOH + Na2CO3  ArONa + NaHCO3
(tannins) (water-soluble)

The tannins extracted from the solid-liquid extraction becomes partially hydrolyzed to form free
gallic acid. Tannins and gallic acid composing of phenolic groups and carboxyl groups are considered
acidic which is treated with sodium carbonate, a base, to convert the acids into their sodium salts that
are highly soluble in water. Caffeine extracted from the tea solution is more soluble in an organic
solvent leaving behind the tannins and the sodium salts of gallic acid in the aqueous layer in liquid-
liquid extraction (LLE). This allows purer caffeine to be extracted from the solution. Boiling chips was
also added to boil the solution in a calm manner on which bumping is reduced, this will prevent the
solvent to boil violently. In solid-liquid extraction the soluble components that includes tannins and
caffeines are isolated in the mixture. However the aim of the study is to extract caffeine and not tannin
and in order to successfully separate those two another method is utilized called liquid-liquid extraction.

LLE or liquid-liquid extraction is one of the oldest method of mixture separation that has a
wide variety of applications in chemical industry , this method is used as a separation technique that
is only possible if the two liquids to be used is immiscible. This process occurs on the transfer of the
desired solute from one solvent to another based on their relative solubilites on the two solvents. The
liquids that comprise these two can be classified as organic phase and the aqueous phase on which
the aqueous phase (polar) and the organic phase (nonpolar) have different preferential solubility
depending on the analyte. From this experiment the solvent to be used is dichloromethane. T the
Dichloromethane (DCM), also called methylene chloride, has a molecular formula of CH 2Cl2 and is a
good extracting solvent but is also toxic. It is a volatile colorless liquid known for its sweet aroma and
is widely used in many industries as an effective organic solvent with its wide range of.The solubility of
caffeine in dichloromethane (140mg/ml) is relatively higher compared to water (22mg/ml) making it a
good extracting solvent (Chaugle et al. 2019). Immiscibility also matters for creating a barrier/layer
caused by the non-polar solvent that exhibits hydrophobic behavior excluding the water molecules
(polar solvent). The differences in density between water and dichloromethane are another factor that
favors DCM. Dichloromethane which possesses higher density compared to water creates an organic
layer below the aqueous layer rather than above like other solvents making it easier to separate the
solvent depleted in solute from the solvent enriched with solute thus minimizing the number of transfer
steps required.

Emulsions are colloids containing two or more heterogeneous types of liquids immiscible
with each other. The two solutions serve as the continuous phase and the dispersed phase where a
cloudy appearance develops producing emulsions. The cloudy appearance is caused by many phase
interfaces scattering light that passes through the mixture at different wavelengths. A basic example of
an emulsion is milk wherein the fat molecules are dispersed in the aqueous solution. The formation of
emulsions is a common laboratory problem encountered during extraction triggered by the presence of
impurities in the solution that act as surfactants or chemical agents reducing the surface tension.
Emulsions are the result of continuous mixing or agitation of two phases. Vigorous mixing in extraction
produces emulsions that may last for a few minutes or longer complicating the separation of the
aqueous solvent from the organic solvent. Swirling the solution slowly in the separatory funnel
minimizes the formation of emulsions thus allowing the transfer of the compound to be extracted into
its more favorable solvent.

When the equilibrium is reached , the chemical potential of the extractable solute is the same
now in the two phases, the compound to be determined is now present in both of liquid layers,
however large amounts of it will be contained on the solvent that has higher affinity , this distribution is
represented by the partition coefficient or the ratio between the concentration of solute at the organic
phase over the concentration of the solute at the aqueous phase since on the experiment
dichloromethane is used the partition coefficient would be 4.6. However, to obtain maximum yield one
must utilize the use of multiple portions than performing a bulk single one. For example if a 5.0 g of
organic compound is dissolved in 100 mL of water and is extracted with three 50-mL portion of ether
(Kd = 10) comparing this with 150 mL portion of ether. The three 50-mL portion would be more
efficient :

With a given KD value of 10, an equation can be made:

 Sorg
K D= =10
S aq

Eq 1. Partition coefficient

If three 50-mL portions are used

Wherein Sorg and Saq is the organic and aqueous phase of organic solvent density.
Furthermore, to determine the amount of compound remaining on both phases within three 50-mL
portion of ether, the given conditions is summarized on the table using Eq 1 where x is the amount
on 1st extraction y as the 2nd initial amount on the second extraction and z as the 3 rd initial amount
on the third extraction.

1st 50-mL ether: 2nd 50-mL ether: 3rd 50-mL ether:

x y z
50 mL 50 mL 50 mL
10= 10= 10=
5−x 0.83− y 0.69−z
100 mL 100 mL 100 mL

4.17 g extracted 0.69 g extracted 0.12 g extracted

0.83 g remained 0.14 g remained 0.02 g remained

Sample calculations for table 4.2

(Initial amount – amount extracted) = amount remained

on the first extraction = 5 – 4.17 g = 0.83 g remained

on the second extraction= 0.83 – 0.69 = 0.14 g remained

on the third extraction = 0.14 g- 0.12 g = 0.02 g remained

Total amount extracted using multiple portions = 4.98 g

If one 150-mL portion is used :

 x
Sorg 150 mL
K D= 10= =4.69 g
S aq 5−x
100 mL

Where x = the amount of extracted in 150 mL portion

From the given solution and data abovementioned, the calculations can be inferred that
the extracted amount using multiple portions is more efficient than using single 150-mL portion.In
addition, the ideal solvent for solvent extraction should fall under these criteria (1) should have a
low boiling point, (2) inert enough to inhibit reactions with solute and solvents (3) not toxic and
highly flammable (4) immiscible with H 2O (5) affordable and should dissolve caffeine at room
temperature. After isolating caffeine from tannins, simple distillation the last method of separation,
will utilize the difference on boiling point , summary on the observations is represented in Table
4.3.

Distillation is a separation method that vaporize a pure liquid and converts it back to liquid
state by cooling in a condenser. There are many types of distillation set ups. On the experiment,
simple distillation is used. Simple distillation is used to isolate two liquids with at least 25 degrees
Celsius difference in boiling points given the liquid should be below 150 degrees Celsius at 1 atm .
This can be used to separate solids and liquids. Another isolation technique is reflux wherein the
condensate is returned to the system where it came from, it also involved condensation of vapors
just like in distillation, the main difference with distillation is that the condenser in reflux is positioned
vertically so the desired liquid remains in the flask as the solvent reaches its boiling point. On the
experiment the dichloromethane extract on which caffeine is dissolved after undergoing liquid-liquid
extraction was filtered through a cotton resulting in a clear solution as residue is removed. After
distilling the dichloromethane , the remaining solution was placed in a pre-weighed evaporation
dish. This evap dish was cooled resulting in a pellucid solution, as a result a needle-shaped crystals
was observed.

Procedure Observation

Filtration of extract Clear solution

Distillation of mixture Clear with minimal solution left

BP range N/A

Evaporation Formation of light yellow-colored crystals

Cooling to room temperature Pellucid solution

 Caffeine crystals Light yellow solid crystals

Table 4.3. Distillation of Mixture.

Parameters Observation

Mass of dried tea leaves (g) 10.005

Mass of evaporating dish (g) 25.673

Mass of evaporating dish + caffeine crystals (g) 25.695

Mass of caffeine crystals (g) 0.022

Percent Caffeine in Dried Tea Leaves (%) 0.220

Table 4.4. Percentage Caffeine in Dried Tea Leaves.

Calculation:

Mass of Caffeine∈grams=( mass of evaporating dish+ caffeine )−Mass of Caffeine

Mass of caffeine ( g )=25.695−25.673=0.022

mass of caffeine ∈grams

%Caffeine ∈Tea Leaves= x 100
mass of dried tealeaves∈ gramas

0.022 g
% Caffeine Tea Leaves= × 100=0.220 %
10.005 g

From a total of 10.005 g of dried tea leaves and the weight of the extract that is 0.022
grams, the percent caffeine in the dried tea leaves is calculated with a mass of 0.220%. This result
signifies that a huge amount of product is lost during the procedure. Same in recrystallization obtaining
a 100% yield is impossible since losses may have already occurred on extraction stages due to
emulsions ,other errors include physical handling that affects the measurement of the weighed crystals,
too much solvent , the way how liquids are mixed during the mixing phase in liquid-liquid extraction and
the way how crystals are formed etc. After performing the experiment one may scrutinized that if an
organic compound can be extracted from an aqueous phase into an organic layer this will lead to 3 to 6
extractions resulting in a tedious procedure ,to minimize the number of steps regarding on this
procedure an organic solvent that is heavier than water can be utilized, for example denser solvents
that includes dichloromethane will remain in the bottom of the separatory funnel while a desired
compound is extracted from aqueous phase, while the solvent that is less dense will remain at the
upper layer of the sep. funnel while extracting a compound in aqueous phase, since the denser organic
solvent can be found on the bottom it can be easily pulled on the solution after extraction without
transferring the aqueous phase from the separating funnel,thus minimizing the steps in extraction
procedure.

Pure caffeine is reported to

melt at 238 °C and sublimes
at a lower temperature thus, one
procedure to obtain the
melting point of caffeine
without sublimating it, a
capillary tube can be utilized
and the temperature can be
moved past 178 degrees
Celsius. Caffeine will melt at
the vapor pressure and the
melting point can be
determined right after heating.
The schematic diagtram for
the separation of four component
mixture that has 4 component
mixture, mainly carboxylic
acid phenol amine and
kethone is summarized in
Figure 4.3 and 4.4
Figure 4.3 Extraction of caffeine
from tea leaves using solid-
liquid extraction and liquid
liquif extractrion

Figure 4.4 Extraction of caffeine from

tea leaves using simple distillation
 Above are the schematic diagrams of the summary of the procedure for extracting caffeine
given the possible compounds on tea leaves such as cellulose, chlorogenic acids, fats, plant
pigments , proteins, soluble peptides, sugars and tannic acid. The tea leaves, together with the
sodium carbonate and boiling chips was dissolved in distilled water and is boiled up to 300 degrees
Celsius at hot plate .Increasing the temperature will increase the solubility making the soluble
compounds such as caffeine and tannins , tannic acid and chlorogenic acid to dissolve in the
solvent. Moreover even if the fats are water insoluble in nature increasing the temperature will disrupt
its hydrophobic structure. During the solid liquid extraction the soluble pigments will also dissolve
allowing it to came in contact with the solvent. Same with the experiment, dichloromethane is used in
order to extract caffeine at the liquid-liquid extraction after adding in the distilling flask, gentle mixing
is performed to prevent the formation of emulsions that will affect the percent recovery of the
experiment, the composition of the mixture that will remain is chlorogenic acid, tannic acids and
sugars, these compounds are insoluble in dichloromethane specifically sugars since they share a
large difference on polarity. On the other hand, the composition of the organic solution would be the
ones that are dissolved by the dichloromethane that includes mainly of caffeine, proteins , peptides
and other impurities as the process goes by the sodium sulphate is added to remove some water
present in the extract this dichloromethane is isolated in caffeine through decantation, filtration and
distillation. After distillation the remaining compounds will be dried crude caffeine. In addition, to
increase the amount of the recovered substance researchers proposed using multiple portion in
extraction instead of using a one bulk of 125 mL solvent.

Extraction is a useful means of separating organic compounds if one compound in the

mixture can be chemically converted to an ionic form. The ionic form is soluble in an aqueous layer
and can be extracted into it. Other, non-ionic organic compounds in the mixture will remain dissolved
in the organic solvent layer. separation of two layers result s in the separation of two
compounds.completion depends upon the relative acidity of the reactants and products. Reactions
occurs so that stronger acid and base are converted into weaker conjugate baseand conjugate
acidsrespectively. The pka value of the acid gives a measure of acidity of each compound. The
separation of the mixtures of a carboxylic acid and phenol would be done using bicarbonate ion since
the carboxylic acid is the only one that is converted to its conjugate base.

CONCLUSION
 On the experiment, there are two main types of extraction that is utilized, first is the solid-
liquid extraction that is used for separating the water soluble components from the insoluble
components of tea leaves and the liquid-liquid extraction, a separation technique that utilizes the
difference of two immiscible liquids to isolate the caffeine using dichloromethane and lastly the
simple distillation set-up that uses the advantages of boiling point difference. After extractions a
total of 10.005 g of dried tea leaves and the weight of the extract that is 0.022 grams, the percent
caffeine in the dried tea leaves is calculated with a mass of 0.220%. This result signifies that a huge
amount of product is lost during the procedure. To conclude a 100% yield is impossible since losses
may have already occurred on extraction stages due to emulsions ,other errors include physical
handling that affects the measurement of the weighed crystals, too much solvent , the way how
liquids are mixed during the mixing phase in liquid-liquid extraction and the way how crystals are
formed etc

References

Rebecca Jeyanthi, Seshiah Candace, Tissopi, Trisha (2014/01/01) “Extraction of Caffeine from used
Tea Leaves, Vol 9” (THE ANNALS OF “VALAHIA” UNIVERSITY OF TARGOVISTE)

Donald L. Pavia, Gary M. Lampman, George S. Kriz, Randall G. Engel (2011) “A Small Scale
Approach to Organic Laboratory Techniques”