Musharraf Abbas Bhatti

Pharm.D, M.Phil
(Pharmaceutics)
Subject – Quality Control
Lahore Pharmacy College
Learning objectives

After attending this lecture you will be able to;

 Definition of alcohol determination

 Methods to determine alcohol contents
 Method I
 Method II
 Method IIA
 Method IIB
ALCOHOL DETERMINATION
 <611> Alcohol Determination

Objective:
―Alcohol determination is used to calculate the amount of alcohol present in in
pharmaceutical materials or glenical prepsrations. ―

 Alcohol can be determined by two methods.

Method I (Distillation Method)
Method II (Gas Chromatographic Method)
 Method II a
 Method II b
 Method I (Distillation Method)
 Method I is to be used for the determination of alcohol, unless otherwise specified
in the individual monograph.
 It is suitable for examining most fluid extracts and tinctures.

Specifications
 Provided the capacity of the distilling flask is sufficient (commonly two to four
times the volume of the liquid to be heated) and the rate of distillation is such that
clear distillates are produced.
 Cloudy distillates may be clarified by agitation with talc, or with calcium
carbonate, and filtered, after which the temperature of the filtrate is adjusted and
the alcohol content determined from the specific gravity.
Precautions
 During all manipulations, take precaution
to minimize the loss of alcohol by evaporation.
 Treat liquids that froth to a
troublesome extent during distillation by
rendering them strongly acidic with
phosphoric, sulfuric, or tannic acid, or
treat with a slight excess of calcium chloride
solution or with a small amount of paraffin
or silicone oil before starting the distillation..
 Prevent bumping during distillation by
adding porous chips of insoluble material
such as silicon carbide, or beads.
For Liquids Presumed to Contain 30% of Alcohol or
Less:
 By means of a pipet, transfer to a suitable distilling apparatus not less than 25
ml of the liquid in which the alcohol is to be determined, and note the
temperature at which the volume was measured.
 Add an equal volume of water, distill, and collect a volume of distillate about
2 mL less than the volume taken of the original test liquid.
 Adjust to the temperature at which the original test liquid was measured, add
sufficient water to measure exactly the original volume of the test liquid, and
mix.
 The distillate is clear or not more than slightly cloudy, and does not contain
more than traces of volatile substances other than alcohol and water.
 Determine the specific gravity of the liquid at 25oC , as directed under
Specific Gravity <841> , using this result to ascertain the percentage, by
volume (ABV), of C2H5OH contained in the liquid examined by reference to
the Alcoholo-metric Table in the section Reference Tables.
For Liquids Presumed to Contain More Than 30% of Alcohol:

 Proceed as directed in the foregoing paragraph, except to do the following:

 Dilute the specimen with about twice its volume of water.
 Collect a volume of distillate about 2 ml less than twice the volume of the
original test liquid.
 Bring to the temperature at which the original liquid was measured.
 Add sufficient water to measure exactly twice the original volume of the test
liquid, mix, and determine its specific gravity.
 The proportion of C2H5OH, by volume, in this distillate, as ascertained from
its specific gravity, equals one-half that in the liquid examined.
Special Treatment:
A) Volatile acids and bases
 Render preparations containing volatile bases slightly acidic with diluted
sulfuric acid before distilling.
 If volatile acids are present, render the preparation slightly alkaline with
sodium hydroxide TS.
B) Glycerin
 To liquids that contain glycerin add sufficient water so that the residue, after
distillation, contains not less than 50% of water.
C) Iodine
 Treat all solutions containing free iodine with powdered zinc before the
distillation, or decolorize with just sufficient sodium thiosulfate solution (1 in
10), followed by a few drops of sodium hydroxide TS.
D) Other volatile substances
 Spirits, elixirs, tinctures, and similar preparations that contain appreciable
proportions of volatile materials other than alcohol and water, such as volatile
oils, chloroform, ether, camphor, etc., require special treatment, as follows:
For Liquids Presumed to Contain 50% of Alcohol or Less:
 Mix 25 ml of the specimen under examination, accurately measured, with
about an equal volume of water in a separator.
 Saturate this mixture with sodium chloride, then add 25 ml of solvent
hexane, and shake the mixture to extract the interfering volatile ingredients.
 Draw off the separated, lower layer into a second separator, and repeat the
extraction twice with two further 25ml portions of solvent hexane.
 Extract the combined solvent hexane solutions with three 10mL portions of a
saturated solution of sodium chloride.
 Combine the saline solutions, and distill in the usual manner, collecting a
volume of distillate having a simple ratio to the volume of the original
specimen.
For Liquids Presumed to Contain More Than 50% of Alcohol:

 Adjust the specimen under examination to a concentration of approximately

25% of alcohol by diluting it with water.
 Then proceed as directed in for Liquids Presumed to Contain 50% of Alcohol
or Less, beginning with ―Saturate this mixture with sodium chloride.
 In preparing Collodion or Flexible Collodion for distillation, use water in
place of the saturated solution of sodium chloride directed above.
 If volatile oils are present in small proportions only, and a cloudy distillate is
obtained, the solvent hexane treatment not having been employed.
 The distillate may be clarified and rendered suitable for the specific gravity
determination by shaking it with about one fifth its volume of solvent
hexane, or by filtering it through a thin layer of talc.
Method II—Gas Chromatographic Method
Method II—Gas Chromatographic Method
Use Method IIa when Method II is specified in the individual
monograph.
For a discussion of the principles upon which it is based, see
Gas Chromatography under Chromatography 621.

USP Reference Standards:

 USP Alcohol Determination—Acetonitrile RS.
 USP Alcohol Determination—Alcohol RS.
Method IIa
Apparatus:
 Under typical conditions, use a gas chromatograph equipped with a
flame ionization detector and a 4mm × 1.8-m glass column packed
with 100 to 120 mesh chromatographic column packing support S3,
using nitrogen or helium as the carrier.
Specifications
 Prior to use, condition the column overnight at 235 with a slow flow
of carrier gas.
 The column temperature is maintained at 1200C and the injection
port and detector temperatures are maintained at 210oC .
 Adjust the carrier flow and temperature so that acetonitrile, the
internal standard, elutes in 5 to 10 minutes.
 Solutions
Test Stock Preparation:
 Dilute the specimen under examination stepwise with water to obtain a
solution containing approximately 2% (v/v) of alcohol.
Test Preparation:
 Pipet 5 ml each of the Test Stock Preparation and the USP Alcohol
Determination— Acetonitrile RS [note—Alternatively, a 2% aqueous
solution of acetonitrile of suitable quality may be used as the internal
standard solution] into a 50-ml volumetric flask, dilute with water to
volume, and mix.
Standard Preparation:
 Pipet 5 ml each of the USP Alcohol Determination—Alcohol RS and the
USP Alcohol Determination—Acetonitrile RS [note—Alternatively, a
2% aqueous solution of acetonitrile of suitable quality may be used as
the internal standard solution] into a 50-mL volumetric flask, dilute with
water to volume, and mix.
Procedure:
 Inject about 5 µL each of the Test Preparation and the Standard Preparation,
in duplicate, into the gas chromatograph.
 Record the chromatograms, and determine the peak response ratios.
 Calculate the percentage of alcohol (%v/v) in the specimen under test
according to the formula:
CD (Ru / Rs)
In which
 C is the labeled concentration of USP Alcohol Determination—Alcohol RS;
 D is the dilution factor (the ratio of the volume of the Test Stock Preparation to the
volume of the specimen taken);
 and Ru and Rs are the peak response ratios obtained from the Test Preparation and the
Standard Preparation, respectively.
System Suitability Test:
In a suitable chromatogram, the resolution factor, R, is not less
than 2;
the tailing factor of the alcohol peak is not greater than 2.0; and
six replicate injections of the Standard Preparation show a
relative standard deviation of not more than 2.0% in the ratio of
the peak of alcohol to the peak of the internal standard.
Method IIb
Apparatus:
The gas chromatograph is equipped with a split injection port
with a split ratio of 5:1, a flame-ionization detector, and a 0.53-
mm × 30-m capillary column coated with a 3.0-µm film of
phase G43.
Helium is used as the carrier gas at a linear velocity of 34.0 cm
per second.
The chromatograph is programmed to maintain the column
temperature at 50oC for 5 minutes, then to increase the
temperature at a rate of 10 per minute to 200 , and maintain at
this temperature for 4 minutes. The injection port temperature is
maintained at 210oC and the detector temperature at 280oC .
Solutions:
Test Stock Preparation:
 Dilute the specimen under examination stepwise with water to obtain a
solution containing approximately 2% (v/v) of alcohol.
Test Preparation:
 Pipet 5 mL each of the Test Stock Preparation and the USP Alcohol
Determination— Acetonitrile RS [note—Alternatively, a 2% aqueous
solution of acetonitrile of suitable quality may be used as the internal
standard solution] into a 25-mL volumetric flask, dilute with water to
volume, and mix.
Standard Preparation:
 Pipet 5 mL each of the USP Alcohol Determination—Alcohol RS and the
USP Alcohol Determination—Acetonitrile RS [note—Alternatively, a 2%
aqueous solution of acetonitrile of suitable quality may be used as the
internal standard solution] into a 25-mL volumetric flask, dilute with water to
volume, and mix.
Procedure:
 Inject about 0.2 to 0.5 µL each of the Test Preparation and the Standard
Preparation, in duplicate, into the gas chromatograph.
 Record the chromatograms, and determine the peak response ratios.
 Calculate the percentage of alcohol (v/v) in the specimen under test
according to the formula:
CD(Ru / Rs)
In which
 C is the labeled concentration of USP Alcohol Determination—Alcohol RS;
 D is the dilution factor (the ratio of the volume of the Test Stock Preparation to the
volume of the specimen taken); and
 Ru and Rs are the peak response ratios obtained from the Test Preparation and the
Standard Preparation, respectively.
System Suitability Test:
 In a suitable chromatogram, the resolution factor, R, between alcohol and the
internal standard is not less than 4;
 the tailing factor of the alcohol peak is not greater than 2.0; and
 Six replicate injections of the Standard Preparation show a relative standard
deviation of not more than 4.0% in the ratio of the peak of alcohol to the
peak of the internal standard.
Reference & Keys

USP 34- NF 29 & BP, 2014

1) <611> Determination of alcohol
Answer the following