EXPERIMENT 2 – SIMPLE AND FRACTIONAL DISTILLATION

Distillation- physical separation process involving the use of selective boiling method and
condensation method.
 Results: complete separation (nearly pure substance) or partial separation
 Increases the concentration of selected components in the mixture. The process exploits the
differences in the volatility of the mixture’s components.
 The formed vapors of the liquid will be the richest in the component of the mixture that boils
at the lowest temperature.
 Purified compounds will boil and will turn into vapors over a relatively small temperature
range.
 The concentration of the lowest boiling component will slowly and steadily deplete, indicating
that a temperature increase is warranted for the next lowest boiling compound. At this point,
the lowest boiling compound has been totally collected and separated from the primary
sample.

Steam Distillation- Method of distilling compounds which are heat-sensitive. The temperature of the
steam is easier to control than the surface of a heating element. This allows a high rate of heat transfer
without heating at a very high temperature

Vacuum Distillation- This is a technique where pressure is lowered in a vacuum. The lowering process
of pressure allows the sample to boil at a lower temperature.

Comparison between Simple and Fractional Distillation

Characteristic Simple Fractional

Boiling Point > 50o C closer boiling points
Apparatus simple distillation set-up With fractionating column
Procedure (to obtain purity) No repetition required Repetition required
Solute/Solvent Separation ✓ ×
Other Uses Purification process for water Petrochemical production
Isolation of Ethanol from Crude Oil refining Process
Fermented Material

Different Types of Alcoholic Beverages, Source and Alcohol Content (Alcohol by volume/ABV)

Type Source Percentage ABV

Vodka Potato Corn Rye or Wheat 40-95
Gin Juniper Berries 36-50
Rum Sugarcane Molasses 36-50
Whiskey Fermented grain mash (barley 36-50
corn rye wheat)
Tequila Blue Agave 50-51
Liqueurs Fruits Herbs and Spices 15
Fortified Wine Grapes fruits berries rice or 16-24
honey + Brandy
Unfortified Wine Grapes fruits berries rice or 14-16
honey
Beer Malted barley 4-8
Malt Beverages Malted Barley 15
Raoult's law- law of thermodynamics established by French chemist François-Marie Raoult in 1887.
 It states that the partial vapor pressure of each component of an ideal mixture of liquids is
equal to the vapour pressure of the pure component multiplied by its mole fraction in the
mixture.
 In consequence, the relative lowering of vapour pressure of a dilute solution of nonvolatile
solute is equal to the mole fraction of solute in the solution.

Dalton’s Law of Partial Pressures- This empirical law was observed by John Dalton in 1801 and
published in 1802 and is related to the ideal gas laws
 It states that in a mixture of non-reacting gases, the total pressure exerted is equal to the sum
of the partial pressures of the individual gases.
𝑎𝑚𝑡 𝑑𝑖𝑠𝑡𝑖𝑙𝑙𝑎𝑡𝑒 𝑜𝑏𝑡𝑎𝑖𝑛𝑒𝑑
% 𝑟𝑒𝑐𝑜𝑣𝑒𝑟𝑦 = × 100
𝑎𝑚𝑡 𝑠𝑎𝑚𝑝𝑙𝑒 𝑢𝑠𝑒𝑑

EXPERIMENT 3 – EXTRACTION

- separation process that consists of the separation compound from a matrix by the use of solvents
which have a high affinity for the compound to be extracted

The distribution of a solute between two phases is considered as an equilibrium condition and is the
basis for partition equilibrium. A solute immersed in two different immiscible solvents will move
towards the solvent that it has a higher affinity towards, as described by Nernst’s Distribution Law.

Selected Compounds Found in Tea

Compound Therapeutic Class Medicinal Uses Structural Formula

Caffeine
(1,3,7-Trimethylxanthine)
Amt in Tea: 11mg/100g tea
Theophylline
(1,3 – Dimethylxanthine)
Amt in Tea: 1mg/L tea
Central Nervous System General Stimulant
Stimulant Bronchopulmonary
Displasia in premature
infants Orthostatic
hypotension
Asthma and COPD Drug Chronic Obstructive
Pulmonary Disease
Asthma
Adenosine Blocker

Theobromine Vasodilator No purported medicinal

(3,7 – Dimethylxanthine) Diuretic use
Amt in Tea: 1.2-3.6 mg/cup Heart Stimulant

Extraction process

a. Separation process: use of an organic compound (solvent that can separate caffeine from the
matrix of tea leaves). Due to the partition theory, extraction selectively dissolves the caffeine
into the solvent used. In the procedure, the use of dichloromethane may include the
extraction of are present in the tea leaves, hence the addition of sodium carbonate is a
necessary step. The sodium carbonate neutralizes the acidic tannins to form insoluble salts
that will remain in the aqueous layer.
 b. Because of the immiscibility of water and dichloromethane, separation of the two solvents is
possible with the use of a separatory funnel due to varying densities.
c. Sodium sulphate: addition of such to evaporated sample is a necessary because excess water
during the separation process may be present. Sodium sulphate acts as a drying agent to
remove the water from the sample obtained after using the separatory funnel, allowing the
collection of pure caffeine crystals.

EXPERIMENT 4 – SUBLIMATION AND MELTING POINT DETERMINATION

Sublimation- physical change of state from solid phase to the gas phase. It is also an endothermic
process that occurs at temperatures and pressures below a substance’s triple point in its phase
diagram which corresponds to the pressure at which the substance can occur as a liquid.

Triple Point- temperature and pressure at which the three phases (gas, liquid and solid) coexist in
thermodynamic equilibrium

Melting Point- temperature at which a solid compound changes to the liquid state. At the melting
point of a substance, equilibrium between the solid phase and the liquid phase is present.

Vapor pressure- pressure exerted by a vapor present above a liquid (solid) surface. As the temperature
of a liquid increases, the kinetic energy of its molecules also increases. As the kinetic energy of the
molecules increases, the number of molecules transitioning into a vapor also increases, thereby
increasing the vapour pressure

Benzoic acid’s triple point is just slightly higher than that of its melting point, 122.37o C, which makes
it bypass the liquid state and directly moves to its gaseous state— Sublimation.

Melting point determination- purity of a sample after separation or extraction procedures. In this
case, a pure substance usually has a sharp melting point or atleast a range that differs by about 1-2o.
Impurities tend to depress/broaden the MP range so that the purified sample should have a higher
and smaller melting point range in comparison to an impure sample. The use of an oil bath or a
sandbath is preferred compared to water because the former compounds accommodate higher
temperatures in comparison to water with a limitation of 100oC.

Methods of Melting Point Determination

Melting Point Apparatus- used to determine the melting point. Some types include the thiele tube,
Fisher-Johns Apparatus, gallenkamp Apparatus, and Automatic Melting Point Apparatus
 A modified version of the thiele tube which was used in the laboratory class may also be used.

EXPERIMENT 5 – RECRYSTALLIZATION

-technique used to purify chemicals by the dissolution of impurities and the compound to be purified
in an appropriate solvent. With this method, the analyst has the option to either remove the impurity
or the desired compound from the solution; dissolution of both impurity and compound to be purified
 Add sample to hot solvent, dissolve and cool
 The pure compound recrystallizes while the impure samples are retained in the solution
 ↓ rate of cooling ↑ crystal development
 The crystals are then collected via filtration and the filtrate is discarded.
Seeding- necessary requirement for the development of crystals in the purification process; addition
of a small amount of the pure compound to the saturated solution or by scratching the glass surface
to create a seeding surface for crystal growth.

Solvent for recrystallization- compound to be purified is not very soluble at low temperatures but
becomes very soluble in higher temperatures; ethanol is a good recrystallizing solvent for acetanilide
due to the fact that acetanilide’s solubility at 0o Celsius is at 18g/mL while at 50o Celsius is 80g/mL.

Acetylation process- signals the addition of an acetyl group to an alcohol or an amine. This is
commonly done in order to place an acetyl protecting group on primary or secondary amines to reduce
their reactivity toward oxidizing agents

Pharmaceutical use of Acetanilide

 First aniline derivative product that possesses analgesic and antipyretic activity
 Market name: Antifebrin but was discontinued due to SE (cyanosis, methemoglobinemia, liver
and kidney damage)
 Metabolite of acetanilide (paracetamol) causes the analgesic and antipyretic effect.
Acetanilide is no longer used as a drug in its own right due to the success of its metabolite.

Limitations of recrystallization
a) Small quantity of impurities
b) Time consuming due to nature of cooling for larger crystals
c) Solvent used should fit appropriately to the sample being purified
d) Amount of impurity > saturation point of solvent @ given temperature: the impurities will not
be filtered out and will remain in the collected crystals

EXPERIMENT 6 – THIN LAYER CHROMATOGRAPHY

- technique used for both qualitative and quantitative identification of compounds and determination
of purity of constituents in a particular sample. It also provides the ability to separate non-volatile
mixtures from each other.

Mobile phase- This is the liquid or gas that flows through a chromatography system, moving the
constituents to be separated at different rates of over the stationary phase

Stationary Phase- The solid or liquid phase of a chromatography system on which the constituents to
be separated are selectively absorbed

rF value- the retention factor (rF) is defined as the distance travelled by the center of the spot to the
distance travelled by the solvent front.

The separation of constituents in a sample is a marker of a constituent’s polarity. The stationary phase
in thin layer chromatography (silica plate) has an adsorptive effect towards polar substances, retaining
them at the spotted area or the spot does not travel too far from the original spot placed. Non-polar
substances have a tendency to go along with the mobile phase (solvent system used) and the most
polar constituents can be found at the highest part of the TLC Plate.
Procedure
1. Prepare 20g of sample and crush in a mortar and pestle; add and mix 30 mL ethanol
2. Place combination in an Erlenmeyer flask and let it stand for 30 minutes with occasional stirring
every 5 minutes
3. In a 250 mL beaker, place about 7mL of the chloroform: methanol solution
4. Cover the beaker with aluminium foil for 15 minutes in order for the solution to equilibriate
5. On a TLC plate, mark both ends of the plate with a line 0.5cm long. Place 2 spots on the bottom
line
6. After 30 minutes, use a capillary tube to obtain a small amount of the sample.
7. Two spots of the sample should be placed unto the TLC plate on the lower line; let it dry
8. Place the lower lined part inside the beaker, letting the silica plate absorb the developing
solution
9. Let the pigments develop up to the higher line; remove plates from beaker and let it dry
10. Place the plates under UV light. Document and observe color spectrum
11. Prepare spray reagent for the plates (vanillin: sulphuric acid) in the sprayer
12. Spray the dried plates with an upward motion from the bottom line

Comparison of Various Chromatographic Procedures

Trait Paper Chromatography Column Chromatography Thin Layer

Chromatograpy
Operational Definition Used to separate colored Used to isolate a single Used to separate non-
substances compound from a volatile compounds
mixture from a mixture
Stationary Phase Cellulose layers in filter Solid material (silica Solid Material (silica
paper gel/alumina) Gel/alumina)
Mobile Phase Organic solvents or Mixture of polar and Mixture of polar and
buffers non-polar solvents non-polar solvents
Elution Compound with Depends upon the Depends on the
increased affinity with solvent system used solvent system for
stationary phase moves (polar = increased separation
slower elution)(non-polar =
retention of polar
substances at the lower
area of the plate)

𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑚𝑜𝑣𝑒𝑑 𝑏𝑦 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑚 𝑠𝑡𝑎𝑟𝑡𝑖𝑛𝑔 𝑙𝑖𝑛𝑒

𝑟𝑓 = ; 𝑢𝑛𝑖𝑡𝑙𝑒𝑠𝑠; 0.0 − 1.0
𝑑𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑚𝑜𝑣𝑒𝑑 𝑏𝑦 𝑖𝑛𝑑𝑖𝑣𝑖𝑑𝑢𝑎𝑙 𝑠𝑝𝑜𝑡 𝑓𝑟𝑜𝑚 𝑠𝑡𝑎𝑟𝑡𝑖𝑛𝑔 𝑙𝑖𝑛𝑒

The Rf (=retardation factor) depends on the following parameters:

a. solvent system
b. absorbent (grain size, water content, thickness)
c. amount of material spotted
d. temperature
 EXPERIMENT 7 – COMPARATIVE INVESTIGATION OF ORGANIC COMPOUNDS

Infrared Spectroscopy- analysis of the interaction between infrared light and a molecule of a particular
compound. This method is used in order to determine the functional groups present in the compound
by the measurement of the vibration of atoms.
Physical properties of Compounds- observable or measurable without changing the composition of a
particular product.
Chemical Properties- observable when a particular object undergoes a chemical change.
Hydrocarbons- Carbon + Hydrogen
Hydrocarbon Derivatives- Carbon + Hydrogen + Another element. Examples include: alcohols, ethers,
amides, carboxylic acids and etc.
Ignition- combustion involving the exothermic chemical reaction between fuel and an oxidant
accompanied by the production of heat and conversion of a chemical species
Solubility- chemical property that refers to the amount of a particular solute that dissolves in a
measured amount of solvent
Intramolecular force- This is any force that binds together atoms, making up a molecule or compound.
Chemical bonds, Ionic bonds and covalent bonds are considered as intramolecular forces
Physical and Chemical Properties of Sample Substances in the Laboratory

Test Cyclohexan DCM Ethanol Phenol Benzoic Ethyl Ethylamin

compound e Acid Acetate e
Molecular C6H12 CH2Cl2 CH3CH2O C6H6O C7H6O2 C4H8O2 C2H7N
Formula H
Physical liquid Liquid Liquid Liquid Solid Liquid liquid
State at
Room
Temp
Appearanc Clear Clear clear Cloudy White Clear Liquid
e
Color Colorless Colorless colorless White White Colorless Colorless
Odor Petroleum- Sweet Alcohol- Sweet Odorless Plastic Ammonia-
like like Balloon- like
like
Solubility in Immiscible Immiscible Miscible Miscible Insoluble Miscible Miscible
H20
in 5% NaOH Immiscible Miscible Miscible Miscible Insoluble Miscible Miscible
In 5% HCl Miscible Miscible Miscible Miscible Insoluble Miscible Immiscible
Reaction Acidic Acidic Acidic Acidic Acidic Acidic Basic
with Litmus
Paper
Ignition Flammable Non- Flammable Non- Non- Flammabl Flammabl
Test Flammabl Flammabl Flammabl e e
e e e