Prepared by:

Hazel
 The odorous, volatile principle
of plant and animal source
are known as volatile oil.
As they evaporate when
exposed to air at ordinary
temperature ,they are also
called ethereal oil.
Volatile oils are also called
essential oil because they
produce essence.
 Chemical components of official volatile oils are:
hydrocarbons, alcohols, aldehydes, ketones, phenols,
acids, and sulfur compounds like allyl isothiocyanate
(in mustard oil);
The purity and quality of volatile oils may be judged to
some extent by their:
- appearance, odor, color,
- and from the determination of the specific gravity,
rotary power, refractive index, solidfying/congealing
point, solubility, and behavior on distillation.
Acetylation Flask Cassia Flask Erlenmeyer Flask
Separatory
funnel

Graduated
Pipette Beaker cylinder
 Sand bath

Water bath

Burette
Electronic hot Analytical
plate Balance
 The acid value is the number of milligrams of
potassium hydroxide (KOH) required to
neutralize the free acids in 1 g of sample.
The acidity may also be determined as the
number of milligrams of 0.1 NaOH required to
neutralize the free acid in 10 g of substance.

 Ester value
The ester value is the number of milligrams
of potassium hydroxide (KOH) required to
saponify the esters in 1 g of sample.


.
 Determination by Acetylation
-the oil is acetylized with acetic anhydride and the ester
content of the resulting oil is determined; from this value the
percentage of alcohol in the original oil may be calculated.

For this determination, a special acetylation flask of

approximately 100ml capacity is employed. This flask is
equipped with an air-cooled condenser attached to the flask
by means of a ground glass joint . A condenser 1 m. in length
is to be preferred in order to prevent the loss of volatile
constituents.
Acetylation
Flask
1. Place 10 ml of the sample in a 100-ml acetylation flask and add 10 ml of
acetic anhydride and 2 g of sodium acetate.
2. Attach an air condenser to the flask and influx the contents gently for exactly
2 hr on a sand bath. Cool the flask for 15 min at room temperature.
3. Add 50 ml of distilled water to the reaction mixture through the air
condenser, and heat the flask on a steam bath for a further 15 min with
frequent shaking.
4. Pour the contents of the fIask into a separatory funnel. Rinse the flask with
two 10-ml portions of distilled water and add the rinsings to the separatory
funnel.
5. Shake thoroughly and allow the mixture to separate into two layers. Reject
the aqueous layer and wash the oil with 100-ml portions of saturated sodium
chloride solution until the washings are neutral to litmus paper.
6. Dry the oil over anhydrous sodium sulfate.

 Assay for Aldehyde Content ( like benzaldehyde,
cinnamic aldehyde, salicyl aldehyde, citral and
citronellal)

Assay for Ketone Content ( like camphor, carvone,

fenchone, thujone, and menthone)
 -This method has proved satisfactory for the
estimation of cinnamic aldehyde in cassia oil, of
benzaldehyde in bitter almond oil, of citronellal in
Eucalyptus citriodora oil; it is the commercially
accepted method for the determination of citral in
lemongrass oil.
The larger flask with a capacity of 150 ml and a
thin neck graduated to contain 6 ml is very
satisfactory for the determination of aldehydes
and ketones. The smaller flask with a capacity of
100 ml and a neck graduated to contain 10 ml
may also be used for such determinations,
although the accuracy will suffer somewhat.
Bisulfite method
1. To a 10-ml sample of the essential oil taken in a 150-ml cassia
flask, add 75 ml of a saturated solution of sodium bisulfite.
2. Heat the flask in a water bath with frequent shaking to dissolve
the solid addition compound and to ensure complete reaction of
the aldehyde with the reagent.
3. Add an additional 25 ml of sodium bisulfite solution and
continue heating.
4. Allow the unreacted oil to form a separate layer.
5. Drive the oily layer into the neck of the cassia flask by slowly
adding sodium bisulfite solution. Measure the volume of the
unreacted oil.
 The aldehyde content may then be calculated
by means of the following formula:

Percentage of aldehyde = 10(10 no. of ml of

unreacted oil).
 The phenols in the volatile oil is absorbed with
solution of KOH, while the non-phenolic
portion of the oil remains undissolved and is
measured in the graduated neck of cassia flask.
Since the potassium salts of many phenols are
more soluble than the corresponding sodium
salts, the use of potassium hydroxide is
preferred.
1. Into a well cleaned 150 ml cassia flask, having a long, thin neck graduated
in 0.1 ml divisions, introduce 10 ml of the oil, measured from a pipette.
2. Add 75 ml of an aqueous 1 N potassium hydroxide solution, measured
from a graduated cylinder.
3. Stopper and shake thoroughly for exactly 5 min.
4. Permit to stand undisturbed for 1 hr., after which the undissolved oil is
forced into the neck by the addition of more KOH solution. The alkaline
solution must be added carefully to avoid disturbing the layer of
separated oil.
5. This addition may conveniently be made by clamping the flask at a slight
angle on a ring stand; above the flask is placed a ring to hold a separatory
funnel containing the solution of alkali which is permitted to flow down
along the inside of the neck of the cassia flask very slowly.
6. If the flow of the alkali is adjusted to about 1 drop per sec, a clean
separation of the oil is usually obtained.
7. In order to make any droplets of oil adhering to the sides of the flask rise
into the neck, gently tap or revolve the flask rapidly between the palms
of the hands. Measure the quantity of oil that does not dissolve in the
alkali. Apparatus For
Phenol
Determination
 The phenol content, expressed as a volume/
volume percentage, is calculated from the
following formula:

Percentage of phenol = 10(10 no. of ml of

undissolved oil)
 The estimation of the volatile oil content of
certain official spirits is based upon the
separation of the volatile oil by means of an
immiscible solvent and measurement of the
volume of the oil.
The apparatus used is a Babcock bottle.
1. The spirit is added with kerosene and saturated calcium chloride solution
acidified with HCl in the Babcock bottle.
2. The acidified calcium chloride solution, and the oil dissolves in the kerosene
when the mixture is shaken.
3. The calcium chloride serves to salt out any volatile oil from the diluted alcoholic
mixture.
4. The acid ensures the absence of alkali which would render acids, traces of
aldehydes, water-soluble.
5. The determination is carried out in a Babcock bottle, the neck of which is
calibrated into eight main divisions of 0.2 mL each.
6. The oil-kerosene layer, being lighter than the aqueous layer, rises into the neck
of the flask when the mixture is centrifuged.
7. The length of the oil-kerosene layer supplies the information needed for the
calculation of the percent of the oil in the spirit.
Example:
If 7.4 divisions represents the length of the column of oil-kerosene in the
graduated neck of the flask
the oil measures 7.4- 5.0= 2.4 divisions, because each 5 divisions are equal to 1ml
( 5x 0.2ml) and 1ml of kerosene was added.
The 2.4 divisions, each of which represents 0.2ml are equal to 0.48ml of oil.
If 5ml sample is employed,100ml of the spirit yielded 0.48 x 20= 9.60ml of the oil.
The procedure is subject to slight error; the volume of oil in scale divisions is
multiplied by 4.2 (20 x 0.21 ml) to correct for the error due to solubility of the oil
kerosene layer in the aqueous layer.
Thus, in the example given, 2.4 x 4.2 = 10.08 ml of oil in 100ml of the spirit.
Thank you