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# Quality Control Lecture

Prepared by:
Perlita Manuzon-Crucis, RPh,MS Pharm
Faculty, College of Pharmacy
GRAVIMETRIC ANALYSIS

##  the measurement of the weight of a substance

in a sample or calculations of the weight of a
substance in a sample from the weight of a
chemically equivalent amount of some other
substances
• small amount of drug is used
 time consuming.
 Precipitation
 Filtration
 Washing
 Drying or ignition to constant weight
Example Assay

##  Assay of Sodium Lauryl Sulfate

Formula:
Sample Problems
 A 0.3056g of sample of a soluble chloride was analyzed
gravimetrically for Cl, and 0.7265g as AgCl was obtained.
Calculate the Cl and the NaCl contents of the sample,
expressing each in %w/w.
Cl = 35.457 Ag = 107.8 Na = 23

##  An unknown sample of asoluble sulfate weighing 1.8

yielded 0.9 g of barium sulfate. Calculate the %S in the
unknown.
MW Sulfur 32 MW BaSO4 233.4
Sample Problems

##  Calculate the amount of each and the

%purity if the unknown in Prob. 1 were
magnesium sulfate, potassium sulfate, and
aluminum sulfate.
ASH CONTENT
 The residue after incineration of the crude drug or drug
from botanical origin is the ash content.

##  It represents the inorganic salts naturally occurring to

the drug and adhering to it and it also include inorganic
matter that is present for the purpose of adulteration.

##  Ash content determination is of importance because it is

a basis for judging the identity and cleanliness of the
drug and gives information relative to its adulteration
ASH CONTENT
 Total ash is the residue remaining after
incineration.

##  Acid-insoluble ash is the part of the total ash

which is insoluble in 3 N HCl.

##  Water-soluble ash is the part of the total ash

that is soluble in water.
Temperature equivalents of an electric
furnace
Description Temperature

## White heat 1200 to 1600°C

Formula:
RESIDUE ON IGNITION (Sulfated Ash)
 Residue on ignition is the measure of the
amount of residual substance not volatilized
from a sample when the sample is ignited in the
presence of sulfuric acid.

##  It also determines the content of inorganic

impurities in an organic substance.

##  Sulfated ash/Sulphated ash found in European

and Japan Pharmacopeia are considered
equivalent to this test, except when noted.
Examples
 Acetazolamide – an upper residue limit of 0.1%

##  Aspirin – an upper residue limit of 0.05%. For this

determination 1 to 2g samples of the official substances
are usually taken.

##  Cocaine hydrochloride – 500mg is taken and the

residue which remains mush not exceed 500microgram
which means form this drug only a negligible amount of
residue may remain

## – Negligible – a quantity not exceeding 500 microgram

Formula:

LOSS ON IGNITION (Loss on Drying)

##  a means of determining the percentage of test

material which is volatized and driven off under
the conditions specified.
Formula:

WATER CONTENT
 Water content of varying quantities of many official
drugs in the USP/NF is either as water of crystallization
or water in the adsorbed form.

##  In order to ensure uniformity for these official drugs, the

USP/NF specify limit of water content on these drugs
OFFICIAL Methods Used to Determine
Water Content
 Method I (Titrimetric)
– method Ia (Direct Titration)
– method Ib (Residual Titration)
– method Ic (Coulometric Titration)

##  Method III (Gravimetric)

ELECTROMAGNETIC TITRATION OF
WATER
 Karl Fisher electrometric titration method – one of the
important procedure in determination of water content
of official drugs.

##  involves the titration of the sample in reagent methanol

with the Karl Fisher reagent which consist of iodine,
sulfur dioxide, pyridine, and methanol.
 Most rapid of the official methods.

##  Widely applicable in determination of the

water content of drug products.
Formula:
Sample Problems
 Compute for water equivalence factor of 180mg of
sample of sodium tartrate (Na2C4H4O6. 2H2O) required
15 ml of Karl Fischer reagent.
Na = 23 C = 12 H=1 O = 16

##  Calculate the %water of the penicillin powder if 9 ml of

Karl Fischer reagent having an F of 3.8 was consumed
by 3.5g sample.

##  Compute for the %Total Ash of a 5.5g sample, If the

weight of the residue after incineration is 0.1211g. The
acid-insoluble ash is 0.1515g.
Seatwork
1. A precipitate of AgCl weighing 0.2432 g was obtained from a 25-
ml sample of a hydrochloric acid solution (ρ=1.2g/mL). What is
the %w/w of HCl content of the solution? (Atomic weights:
Ag=108, Cl=35, H=1)

## 2. The residue after incineration of a crude sample was 0.1152g

and the residue after treatment of the ash with diluted HCl was
0.0651g. Calculate the %silica obtained from 7.8525g sample.

## 3. Solve for the %LOl of hydrated MgSO4 if 3.2g sample gave a

residue after ignition of 2.15g.

## 4. Calculate the %sulfated ash of 3.5g plant sample after addition

of sulfuric acid gives a residue of 0.1609g. The weight of the
silica in the sample is 0.1718g.
Sample Problems
EXTRACTIVE AND CRUDE FIBER
CONTENT
 The amount of extractives a drug or a substance yield
represents the amount of constituents or group of
related constituents the sample contains.

##  In some cases, the amount of a drug soluble in a given

solvent is a measure of purity.
VOLATILE AND NONVOLATILE ETHER-
SOLUBLE EXTRACTIVE

##  The determination of total ether-soluble volatile

constituents is applied to drugs which contain volatile
oils, while the determination of nonvolatile ether-
soluble constituents is applied to drugs having active
constituents associated with volatile matter.

##  Volatile substance are volatilized leaving resin, coloring

matter and fixed oil as non-volatile ether soluble
content.
ALCOHOL-SOLUBLE EXTRACTIVE
 Alcohol – good solvent for resinous matter.

##  Determination of alcohol-soluble extractive –

most frequently employed to determine
approximately the amount of resin in drugs in
which resinous matter is the important
constituent.

##  Method I – hot extraction

 Method II – cold extraction method
WATER-SOLUBLE EXTRACTIVE AND
WATER-INSOLUBLE RESIDUE

##  Determination of the water-soluble extractive content –

applied to drugs of which one or more of the most
important constituents are soluble in water.
SOLVENT HEXANE-SOLUBLE
EXTRACTIVE

##  Hexane extractive – an index in the amount of fat

present and serves as a check on the amount of seeds
contained in colocynth pulp, since the seeds contain a
large amount of fatty matter
CRUDE FIBER
 the residue, consisting chiefly of cellulose, that remains
undissolved after successive treatment with boiling acid
and alkali.

##  important in the examination of certain drugs and

particularly of spices, since the commonly used
adulterants consist of waste or refuse material derived
from the drugs or spices themselves or from other food
products.

##  frequently this material is the outer cellular layer or

protective coating, which contain a larger proportion of
lignified tissue and consequently, crude fiber.
CONSTANTS OF FATS, FATTY OILS,
WAXES, BALSAMS, AND RESINS
 The methods of analysis offatty substances, waxes,
resins, etc., usually consist of the determination of a
number f physical and chemical properties or values
commonly known as constants.

##  These constants, when taken in conjunction with color,

odor, taste, and special identity tests for given
substances and for common adulterants, are the basis
upon which the purity and quality of these substances
are judged.

##  Specific gravity, solubility, melting point, RI, and optical

activity.
Acid value
 a.k.a. acid number and acidity index.

##  the number of milligrams of potassium hydroxide

necessary to neutralize the free acid in 1g of oil, fat,
wax, resin, balsam, or similar organic substances of
complex composition.

##  Phenolphthalein – indicator used

 For the titration, 0.5N, 0.1N or 0.02N of alkali may be
employed but a solution of 0.1N is the most suitable in
the majority of official determinations.
Formula:

## V x N 56.11 mg/meq KOH

A.V. = -----------------------------------
Sample weight
Sample Problems:
 If a 2g sample of cod-liver oil required 4.5ml of 0.02N
NaOH in the titration of the free fatty acids, would be
the oil conform with the official purity requirement?
What would be the acid value of the oil?
K = 39.1 O = 16 H=1
 Cod-liver oil specs - amount used (2g), alkali used 0.1N
NaOH, Official req’t (≤1ml)

##  If a 2108 mg sample of castor oil required 17.5ml of

0.02N NaOH in the titration of the free fatty acids, would
be the oil conform with the official purity requirement?
What would be the acid value of the oil?
Saponification Value
 a.k.a. saponification number and Koettsdorfer number

##  the number of milligrams of potassium hydroxide

required to neutralize the free acids and soponify the
esters contained in 1g of fat, fatty or volatile oil, wax,
resin, balsam, or other substances of similar
composition.

##  serves to aid in the detection of the presence of the

glycerides of acids containing less than 16 or more than
18 carbon atoms, since the value of this constant is
inversely proportional to the mean molecular weights of
the acids present.

##  it may also indicate adulteration of the sample with

unsaponifiable matter, such as mineral oil.
Formula:

## (ml blank – ml sample) (N HCl) 56.11mg/meq KOH

S.V. = ---------------------------------------------------------------
Sample weight
Sample Problem:

##  A sample of corn oil weighing 1600g saponified with

25ml of 0.4N KOH required 9 ml of 0.5N HCl to titrate
the excess KOH. In the blank determination, 20ml of
0.5N HCl was required to titrate the alkali. Calculate the
saponification value of the sample. Does the value
correspond to the U.S.P. requirement?Specs 187-193
Ester value
 the number of milligrams of potassium
hydroxide required to saponify the esters in 1g
of a fatty or volatile oil, fat, wax, balsam, resin
or similar substances.

##  important in the analysis of yellow and white

wax, since it serve in many cases, to indicate the
presences of adulterants, such as paraffin.

##  may aid in the detection of such adulterants as

rosin and stearic acid in the wax.
Formula:

##  Saponification value = Ester value + Acid

value
Unsaponifiable Matter
 the substances present in oils or fats that are not
saponified by alkali hydroxides but are soluble in
ordinary fat solvents.

##  When oils and fats are saponified, there remains a small

amount of residue that may consist of phytosterol in
vegetable oils and fats, of cholesterol in animal oils and
fats.
Formula:

Wt. of residue
%U.M. = ----------------- x 100
Wt. of sample
Iodine value

##  the number of grams of iodine absorbed under

specified conditions by 100g of oil, fat, wax, or other
substances.
 quantitative measure of the proportion of unsaturated
fatty acids present, both free and combined as esters,
that have the property of absorbing iodine.

##  not only serves as an aid in the identification of known

oils but also serves to indicate in a definite manner the
class to which an unknown fat or oil belongs
Iodine value
 Drying oils – such as linseed oil, and fish oils, such as
cod-liver oil, have very high iodine numbers, usually
above 120 since they contain a large proportion of
unsaturated fatty acids.

##  Nondrying oils – such as olive oil and almond oil, have

relatively low iodine numbers, below 100.

##  Semidrying oils – such a s cottonseed oil and sesame oil,

have intermediate iodine values, between 100 to 120.

##  Animal Fats – the iodine number is not very high, usually

being less than 90.
Methods of Determination
 Hanus method – Method 1
– Chloroform (organic) is used because it is soluble in
organic solvent.
– Iodobromide TS is used, instead of Iodochloride

##  Wijs method – Method II

– Iodochloride TS is used
Formula:

## (ml blank – ml sample) (N Na2S2O3) (meq. of

iodine)
Iodine Value = ---------------------------------------------- x
100 Weight of sample
Sample Problems:
 Determine the iodine value of the unknown
sample weighing 0.25g, if 24.15ml and 12ml of
0.1055N sodium thiosulfate was used in titration
with blank and sample respectively.
I = 126.9

##  Calculate the iodine value of talisay seed oil

using 0.25g of the oil as a sample. The oil
consumed 7.2 ml of 0.1095N Na2S2O3 while the
blank titration at the sample condition used 24.8
ml of the sample thiosulfate solution. Calculate
the I.V. of the sample.
Hydroxyl Value
 a.k.a. hydroxyl number

##  the number of milligrams of potassium hydroxide

equivalent to the hydroxyl content of 1g of the
substance.

##  gives an indication of the identity and purity of fatty

substances possessing alcoholic hydroxyl groups

##  An abnormally low value is indicative of adulteration

with higher-molecular-weight alcohols or with
nonalcoholic fatty substances (e.g. paraffin, petroleum
oil)
Acetyl value of Fatty Acids
 the number of milligrams of potassium hydroxide
required to neutralize the acetic acid obtained by the
saponification of 1g of acetylated fatty acids.

##  - corresponds closely to the hydroxyl value of fatty

alcohols, and the two constants have much the same
significance with respect to identity and purity of the
substances.
Water and Sediment in Fatty Oils
 certain fatty oils, particularly those of animal
origin, which are not highly refined, frequently
contain some moisture and nonfatty tissue
residue.

##  limit for moisture and sediment;

nondestearinated cod liver oil – nmt 0.5% by
volume of water and sediment

##  pear-shaped centrifuge with graduation.

ASSAY OF VOLATILE OILS

##  Volatile Oils – a.k.a. ethereal oils, essential oils, or

essences
CHEMICAL COMPONENTS OF OFFICIAL
VOLATILE OILS
 Hydrocarbon – pinene, camphene, limonene
 Alcohols – borneol, linalol, methol
 Aldehydes – benzaldehyde, cinnamic aldehyde,
citral
 Ketones – camphor, carvone, menthone
 Phenols – anethol, eugenol, thymol
 Acids – propionic, butyric, bornyl acetate, mentyl
acetate
 Sulfur compounds – allyl isothiocyanate
General tests
 Sp gravity
 0.84 – 1.2
 Lighter than water(orange, caraway,
lemon, turpentine, rosemary) – HC,
alcohol, esters, ketone
 Heavier than water (anise, cinnamon,
clove, sassafras) – aldehydes, phenol,
phenolic derivatives, certain esters
General tests
 Rotatory power –
 Means of determining adulterant lemon oil
(+57 to +65), with turpentine oil (+25 to -
40)
 RI –1.46 and 1.61
 Water – 1.3333 at 20 deg; 1.3325 at 25
deg
 20 deg
 Rose oil – 30 deg
 Congealing temperature –
 Inc – valuable – anise, eucalyptus – large
amount of crystallizable constituents
 Solubility
Assay of Ester Content (Assay of
peppermint oil for total esters)
 The esters of peppermint oil, which consist chiefly of
menthyl acetate, are saponified by the alkali when the
mixture is refluxed, forming free menthol and potassium
acetate.

##  alcoholic KOH is used, since oil is soluble in strong

alcoholic solution.
Formula:

## (ml blank – ml sample)HCl (N.F. HCl) (99.15 titer

of methyl acetate)
%menthyl acetate or total ester = -----------------------x 100
C12H23O2 Sample weight (mg)
Sample Problem:

##  Calculate the %menthyl acetate if a 9.120g

sample of a peppermint oil, refluxed with 25ml
of about 0.5N alcoholic KOH, required 21.52ml
of 0.4900N HCl for the residual titration. The
blank was run using the same volume of 0.5N
alcoholic KOH and require 26ml of 0.4900N HCl
to bring about the end point.
Assay for Alcohol Content (Assay of
Peppermint Oil for Menthol)
 the acetylization flask should be supported on an
asbestos board while heating to prevent decomposition.
 The acetic anhydride reacts with the menthol, forming
menthyl acetate and acetic acid.
 The acetylization is promoted by the anhydrous sodium
acetate.
 The oil is washed with diluted sodium carbonate TS
(0.6g of Na2CO3 in sufficient distilled water to make
100ml) to neutralize the excess acetic acid.
 The 0.5N KOH saponified the menthyl acetate, forming
menthol and potAc.
 The liberated menthol dissolve in the alcoholic solution.
 nlt 50% of total menthol C10H19OH
Formula:

7.813 A (1-0.0021E)
%total menthol = ----------------------------
B – 0.021A

## where: A = ml blank – ml sample

B = wt. of acetylated oil
E = %total ester
Assay of Aldehyde Content (Assay of
Benzaldehyde)
 When no other constituents are present which
react with the reagent, the assay of aldehydes
may be performed by the bisulfite method
(NaHSO3).

##  The bisulfite addition product dissolves in water,

leaving the nonaldehyde constituents as a water
insoluble layer – measured in cassia flask,
fuses to the surface and when the flask is filled,
this oily layer rises into the graduated neck of
Assay of Aldehyde Content (Assay of
Benzaldehyde)

##  Volatile oils which contain aldehyde in very small

amounts, such as lemon oil, or which contain other
constituents that form water-soluble addition products
with sodium bisulfite cannot be assayed accurately by
bisulfite method instead by the hydroxylamine
method.
Formula:

## Vol. of sample – vol. of water (insoluble layer)

%aldehyde = ------------------------------------------------x 100
Vol. of sample

## (ml blank – ml sample) N.F. x 106.1

mg/meq. (titer of benzaldehyde)
%benzaldehyde = ------------------------------------------x 100
Sample weight (mg)
Sample Problem:

##  In phenol content determination of a volatile oil,

the layer in the graduated cylinder neck of the
of 10ml of the oil after treatment with KOH
solution. Calculate the % phenol in the sample
Assay of Ketone Content (Assay of
Caraway Oil)
reaction or by the hydroxylamine method.

##  Carvone – the principal ketone present in oil, reacts with

the sodium bisulfite.

##  Another official oil evaluated for its ketone content is

spearmint oil. It is assayed in exactly the same way as in
caraway oil, and it is required to contain not less than
55.0% by volume of carvone.
Assay of Phenol Content (Assay of
Clove Oil)

##  Volatile oils that contain phenols when shaken with

solutions of sodium hydroxide diminished in volume
because of the ready solubility of the phenol
constituents in alkali; the nonphenolic portion of the oil
ASSAY OF ALKALOIDS AND AMINE
DRUGS
 chemical substances which are (1) obtained from plant,
animal, or synthetic resources, (2) contain organic
nitrogen(s) within their chemical structure, and (3)
usually possess physiological activity.
 Nitrogen(s) in alkaloids – is usually a primary, secondary
or tertiary amine the type formula R3-N will be used to
represent amines and amine-type alkaloids.
 Amine nitrogen – possess basic properties, hence have a
free pair of electrons which results with hydrogen ions in
acidic substances to form coordinate covalent bonds and
substances chemically classified as salts.
Alkaloidal Test Solutions
 mercuric iodide TS - commonly known as Valser’s
reagent.
– this reagent forms white precipitate with minute
traces of many alkaloids when added to their acidified
aqueous solutions.
 Iodine TS – commonly known as Wagner’s reagent.
– yields reddish or red-brown precipitates.
 Mercuric Potassium Iodide TS – known as Mayer’s
reagent
– yields white or slightly yellow precipitates
Proximate and Ultimate Assays
 Proximate assay – the percent of extractive
represents the total of a class of plan principles,
such as alkaloids or glycosides, etc.

## – Assay of Ipecac – an example of proximate

assay because the total ether-soluble in
alkaloidal content is determined.

##  Ultimate Assay – the percent of extractive from

a crude drug represents a single chemical
species, such as morphine.
Assay of Pharmaceutical Dosage Forms
for Alkaloidal content(official) Assay of
Ephedrine Sulfate Injection
(C10H15NO)2 .H2SO4
 6-50mg/mL vial of ephedrine sulfate, CHCl3, NaCl,
NaOH TS
 0.1N perchloric acid – titrant used.
 428.56/2000
 Titer 21.43 mng
 Specs – 98-101%
Assay of Aminophylline Tablets
(Residual Precipitation)
 Aminophylline – 2 mole of theophylline
(C7H8N4O2)2
 1mole of ethylenediamine (C2H8N2)
 Standard solutions used – AgNO3 and
NH4SCN
 Titer 21.02 mg
 Specs 93-107%
Formula:
(N.F. x ml) AgNO3 - (N.F. x ml) NH4SCN x
(21.02) titer of aminophylline
%aminophylline = -------------------------------------------- x
100
Sample weight
Sample Problem:

##  Calculate the %w/w of aminophylline if a

0.3120g sample, assayed following the
procedure given, required 12.10ml of 0.100 N
NH4SCN to bring about the end point in the
residual titration of AgNO3. Exactly 20ml of
0.1200N AgNO3 was taken initially.
Seatwork
1. Determine the iodine value of a sample of olive
oil weighing 0.2100 g if 24.15 mL and 12.00 mL
of 0.11 N Na2S2O3 which are required for the
blank and residual titration, respectively.

## 2. Find the acid number of a rosin sample

weighing 1.10 g which required 28.00 mL of
0.11 N NaOH to bring about the end point.

## 3. If a sample of beeswax is found to have an acid

number of 15.50 and saponification value of
71.20, the ester value of the sample is
.