7.

0 SPECIAL METHODS OF ANALYSIS OF CRUDE DRUGS

Ms. Melissa Calambro || October 2022 PHARCHEM3
Transcribers: Navarro
Editors: Montepio

volume(mL)
Special methods of analysis %H2O = × 100
weight of sample(g)

WATER CONTENT DETERMINATION TITRIMETRIC

 Official methods:
o Gravimetric  Also known as Karl Fischer Method
o Azeotropic  Reagent: Karl Fischer Reagent
o Titrimetric o Pyridine, Iodine, Sulfur Dioxide(Anhydrous), Methanol
o Dew point  Primary standard: Sodium tartrate
o Electrolytic hygrometric
volume (mL) of KFR x
%H2O = × 100
LIMIT TEST weight of sample (mg)
o Ash Content
o Crude Fiber WEF/Water Equivalence Factor = 0.1566 mg
o Residue on Ignition
o Loss on Ignition LIMIT TEST
Test designed to identify impurities which are likely present in
TOTAL EXTRACTIVE DETERMINATION the sample
o Ash Content
Constants of fats, fixed and volatile oils, waxes, o Crude Fiber
balsams and resins o Residue on Ignition
o Acid Value o Loss on Ignition
o Ester Value
o Saponification Value ASH CONTENT
o Hydroxyl Value
o Acetyl Value of Fatty Acids  Residue remaining after incineration at dull red heat
o Iodine Value or Number (550-700 degree Celsius)
o Unsaponifiable Matter  Ash determination furnishes a basis for judging the identity
and cleanliness of a drug and gives information relative to
ASSAY of Volatile oils/ethereal oils/essential oils its adulteration with inorganic matter.
o Assay for ester content  Types:
o Assay for alcohol content o Total ash
o Assay for aldehyde content o Acid-insoluble ash
o Assay for ketone content
o Assay for phenol content  Total ash – represents the inorganic salts/ substances
o Assay for volatile oil in spirits present (residue remaining after incineration)

WATER CONTENT DETERMINATION weght of total ash

%TA = × 100
initial wt (g)
 It is also known as aquametry
 It is the determination of the amount of water of  Acid-insoluble ash - the part of the total ash which is
crystallization or water in adsorbed form. insoluble in diluted HCl or 3N HCl.
 Official methods: The ash represents silica.
o Gravimetric
o Azeotropic weght of AIA(silia)
o Titrimetric %AIA = × 100
initial wt (g)
o Dew point
o Electrolytic hygrometric
Temperature Equivalents
500-550 deg C Very dull red heat
GRAVIMETRIC 550-700 deg C Dull red heat
 Also known as drying or thermal method 800-1000 deg C Bright red heat
1000-1200 deg C Yellow red heat
 Equipment: desiccator, oven, moisture balance
1200- 1600 deg C White heat
 Two types:
o Gravimetric for drugs containing no constituents, other
than water, volatile at 105 degree Celsius
CRUDE FIBER
o Gravimetric for drugs containing ether-soluble
constituents volatile at 105 degree Celsius  Residue that remains undissolved after successive
Loss in weight of sample treatment with boiling acid and alkali
%H2O =
initial weight of sample  Represented chiefly by cellulose

AZEOTROPIC weght of crude fiber (cellulose)

%CF = × 100
initial wt (g)
 Also known as Toluene distillation or Xylene method
 An azeotropic is a mixture with a constant boiling point. Weight of crude fiber = weight of residue = weight of ash
 Equipment: Toluene Moisture Tube
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[PHARCHEM3] 7.0 SPECIAL METHODS OF ANALYSIS OF CRUDE DRUGS – Ms. Melissa Calambro
RESIDUE ON IGNITION volume of 0.5 N alcoholic KOH and required 26.00 mL of
0.5100 N og HCl to bring about the end point.
 This is necessary to control the amount of inorganic matter
Answer: 192 mg KOH / Saponification Value of the oil
that may be contained in such substances as alkaloidal
salts, especially when the substance is intended for
2. If a sample of white beeswax has an acid value of 18.45
hypodermic administration.
and an ester value of 74, what is the saponification value
of the sample?
LOSS ON IGNITION
Answer: 92.45
 Provides a means of determining the percentage of test
material which is volatilized and driven off under the 3. If a sample of white wax is found to have an ester value of
conditions specified. 65.7 and a saponification value of 74.2, what is the acid
value of the sample?
TOTAL EXTRACTIVE DETERMINATION a. 8.5
Extraction is the process of removing plant/animal constituents b. 86.5
by the used of a suitable solvent (menstruum) and process c. 186.5
d. 56.5
weght of crude extract
%Extractive = × 100 Hydroxyl Value
initial wt of sample (g)
number of mg of KOH equivalent to the hydroxyl content of 1 g
o Soxhlet apparatus – used for the extraction with of substance. This gives an indication of the identity and purity
volatile solvents where small quantities of a drug are of fatty substances possessing alcoholic hydroxyl groups.
extracted.
o Alcohol – resinous matter Acetyl Value of Fatty Acids
o Hexane – fats and fatty oils number of mg of KOH required to neutralize the acetic acid
obtained by the saponification of 1 g of acetylated fatty acids.
SAMPLE PROBLEM
The % hexane extractive obtained from 27.5820 g of crude drug Iodine Value or Number
yielding a residue of 0.9155g of extractive is:
a. 3.32% number of g of iodine absorbed under specified conditions by
b. 33.2% 100 g sample. This provides a quantitative measure of
c. 4.30% unsaturated fatty acid.
d. 4.6% Method I – Hanus Method – uses Iodobromide TS
Method II- Wijs Method – uses Iodochloride TS
Constants of fats, fixed and volatile oils, waxes, balsams (volume blank − volume actual) × N Na2S2O3 × 0.1269
and resins �� = × 100
Weight of sample

Value/Acid Number/Acidity Index SAMPLE PROBLEM

the number of mg of KOH necessary to neutralize the free Determine the iodine value of an unknown sample of oil
acids in 1 g of sample. weighing 0.21g if 26mL and 12mL of 0.1100 N of sodium
� × � × ��. �� thiosulfate are required for the blank and residual titration
�� = respectively.
�
a. 90
Ester Value/ Ester Number b. 93
c. 108
the number of mg of KOH required to saponify the free esters in d. 200
1 g of sample (fatty or volatile oil, fat, wax, balsam, resin, or
similar substance. Iodine Value
� (����� − ������) × � ��� × ��. �� Oil IV
�� =
� DRYING OILS Very high iodine values
 LINSEED OIL Large proportion of
Sample Problem  FISH OIL/COD OIL unsaturated FA
If a 2 g sample of cod liver oil required 4.5 mL of 0.02 N KOH in Above 120
the titration of the free fatty acids, what would be the acid SEMI-DRYING OILS Intermediate iodine values
 COTTONSEED OIL 100-120
number of the oil?
 SESAME OIL
Answer: 2.52 NON-DRYING OILS Relatively low iodine
 OLIVE OIL numbers
Saponification Value/ Saponification Number/Koettsdorfer Number  ALMOND OIL Below 100
number of mg of KOH needed to saponify the free esters and
neutralize the free acids in 1 g of sample.
Unsaponifiable Matter
� (����� − ������) × � ��� × ��. �� defined by the USP as the substance present in oils or fats that
�� =
� are not saponified by alkali hydroxides but are soluble in
SV= EV + AV ordinary fat solvents.
Sample Problems:
1. Find the saponification value of cotton seed oil if a 1.532-g
sample refluxed with 25.00 mL of about 0.5 N alcoholic
KOH, required 15.70 mL of 0.5100 N of HCl for the
residual titration. The blank was run using the same
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[PHARCHEM3] 7.0 SPECIAL METHODS OF ANALYSIS OF CRUDE DRUGS – Ms. Melissa Calambro

ASSAY of Volatile oils/ethereal oils/essential oils  Assay of peppermint Split

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, solidifying/congealing point, solubility and behavior on
distillation.
o Assay for ester content
o Assay for alcohol content
o Assay for aldehyde content
o Assay for ketone content
o Assay for phenol content
o Assay for volatile oil in spirits

Assay for ester content

Volatile oil are evaluated on the basis of their ester content.

Assay for alcohol content

The total alcohol present in any given oil are determined by
transforming the free alcohols into the corresponding acetates
by boiling the oil with acetic anhydride in an acetylation flask
(see Figure 1) and then determining the saponification value of
the acetylized product.

Assay for Aldehyde content

 When no other constituents are present which react with
the reagent, the assay for aldehydes may be performed by
the bisulfite method.
 Aldehyde form addition products with certain reagents and
this product dissolves in water leaving the non-aldehyde
constituents as a water insoluble layer. The volume of this
water-insoluble layer is then measured in a cassia flask.
 Assay of Benzaldehyde

Assay for Ketone Content

 The determination is made by bisulfite addition reaction or
by the hydroxylamine method.
 Assay of Caraway Oil for the presence of Carvone

Assay for Phenol Content

 Volatile oils that contain phenol when shaken with solution
of hydroxide diminish in volume because of the ready
solubility of phenol constituents in alkali. The non-phenolic
portion of the oil remains undissolve.
 In this determination, a cassia flask is used. (see Figure 2)
 Assay of Clove Oil for the presence of Eugenol

� ������ − �������
%������ = × ���
�������

SAMPLE PROBLEM
In phenol content determination of a volatile oil, the layer in the
graduated neck of the cassia flask read 2.3 mL obtained from a
sample of 10 mL of the oil after treatment with KOH solution.
The % phenol is:
a. 73
b. 69
c. 7.3
d. 77

Assay for Volatile Oil in Spirits

 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 Bobcock bottle. (see Figure 3)

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