Special methods

Ash content Determination

Ash Value
• residue remaining after incineration is the ash content of the drug.
• Inorganic salts of carbonates, phosphates, silicates of sodium, potassium, calcium and
• magnesiu .a criterion to judge the identity or the
purity of the crude drug.
A high ash value is indicative of contamination, substitution, adulteration or carelessness in preparing the crude drug for marketing.
– is used as a basis for the identity and cleanliness of a drug.
– This also gives information about adulteration with inorganic matter.
• The ash or residue yielded by organic matter is a measure of the amount of inorganic matter
present as impurity.
– used to measure the total amount of material remaining after incineration
– is the residue obtained after boiling the total ash with dilute HCl and igniting the remaining insoluble matter
– is the difference in weight between total ash and residue after treatment of total ash with water
– careful control of temperature is the most important analytical factor to regulate.
When using Electric furnace for ignitions, the following temperature equivalents should be considered:
a) Very Dull Red Heat – 500-550 C
b) Dull Red heat – 550 – 700C
c) Bright Red Heat – 800-1000C
d) Yellow Red Heat – 1000-1200 C
e) White Heat – 1200 – 1600C
Different source of ash of a drug:
Carbonates
Phosphates
Sulfates
Chlorides
Oxides of Ca, Mg, K, Na, Al, Fe, and other metallic elements
 Add Filter
Total 10mL through Ignite Cool
water ashless at and
Ash and boil filter 450˚C weigh
for 5 paper

Weight of Water-soluble ash = weight of total ash – weight of water insoluble ash
 Sample Problem
The following data were obtained after the moisture
and total ash content determination of acacia powder:
Weight of empty crucible – 62.45 g
Weight of crucible with sample – 71.65g
Weight of crucible with sample after drying to constant
weight – 70.50g
Weight of crucible and residue left after incineration –
63g

a. What is the moisture content in grams?

b. Percent moisture of the sample?
c. What is the total ash content in grams?
d. Percent Total ash?
• METHOD I: Karl Fisher Titrimetry
– 1mL of KFR = 15mL H2O
• METHOD II: Azeotropic Distillation (USP) / Moisture
method by toluene distillation (NF) – Xylene
– Moisture is removed from the sample by distillation as an
method
azeotrope (constant boiling point mixtures) with a solvent.
• METHOD III: Gravimetric Method – Method of
Choice -Dried at 105˚ C for 5 hours
a) For drugs containing no constituents, other than water,
volatile @ 105˚ C
b) For drugs containing ether- soluble constituents volatile @ 1 0 5 ˚
C
• Dew point
• Electrolytic hygrometric
• The amount of extractive is often an
approximate measure of the amount of certain
soluble constituents or group of related
constituents within a crude in a certain solvent.

• The solvent to be employed must not dissolved

appreciable quantities of substances other than
those sought In the extraction

• An efficient and complete extraction is assured if

the solvent is divided into a number of relatively
small portions and used successively.
Soxhlet Apparatus
 Alcohol – Soluble Extractives
• Alcohol is an ideal solvent for
extraction of various chemicals like
tannins, resins

• Alcohol – Soluble extractives is the

most frequently employed to determine
the
approximate amount of resin in drugs

• 95% ethyl alcohol is used for

determination of alcohol soluble
extractive
 Water-soluble extractive
and Water-insoluble
residue
• The determination of the water soluble
extractive content is applied to drugs
of which one or more of the most
important constituents are soluble in
water.

• The determination is performed in

the same way as the determination
of the diluted alcohol soluble
extractive.
 Crude Fiber
• This is the residue, consisting chiefly of cellulose, that
remains undissolved after successive treatment with boiling
acid and alkali.

Crude fiber content as a percentage of the sample:

= B-C x 100/ A
A = mass of sample in grams
B= loss of mass after ashing during determination C= loss of
mass after ashing during blank test
The difference between two
paralleldeterminationscarriedout on the sample must
not exceed0.3 inabsolute value for
crude fiber contentslower than10%.
Crude fiber content % = 100 (A-B/C)

A= wt. of crucible with dry residue in

grams
B= wt. of crucible w/ ash in
grams C= wt. of sample in grams
 Constants
• Are values which are used as
parameters to measure the physical and
chemical
properties of fatty substances, fatty
oils, waxes, balsams and resins.

• used as a basis to determine the

purity and quality of these substances
Common constants used in measuring
quantitatively the properties of fatty substances:
• Acid value
• Saponification
value
• Ester value
• Iodine value
• Hydroxyl value
• Acetyl value
 Acid value
• Also known as acid number or acidity index

• The number of mg of KOH necessary to

neutralize the free acids in 1 g of oil, fats
wax, resin, balsam or similar organic
substance

AV = N x V x 56.11
wt. in g
Sample Problem
• Find the acid number of a rosin sample
weighing 1.1g which requires 28 ml of
0.11N NaOH to bring about the end point.
 Saponification value
• Also known as saponification number or
Koettsdorfer index
• Saponification is a process by which triglycerides
are reacted with sodium or potassium hydroxide
(lye) to produce glycerol and a fatty acid salt called
"soap."
 Saponification value
• The number of mg of KOH necessary to
neutralize the free acids and saponify the
esters in 1 g of oil, fats wax, resin, balsam or
similar
organic substance.

SV = AV + EV
= [(Vb-Va) N x 56.11]
wt in g
 Sample Problem
Find the saponification value of cottonseed
oil if a 1.532g sample is refl uxed with 26ml
o f 0. 5 N a l c o h o l i c K O H , r e q u i r e d 15. 70 m l o f
0.51 N HCl for the residual titrat ion . The
blank was run using the same volume of
0.5N Alc . KOH and requires 26ml of 0.51 N
HCl to bring about the end point.
 Ester value
• Also known as Ester number

• The number of mg of KOH require to saponify the esters in

1 g of volatile oil, fats wax, resin, balsam or similar
organic substance.

• Substances that contain free fatty acids , the

ester value is equal to the saponification number.

• Ester value is the difference between the

saponification and acid number.
• Important in the analysis of yellow and white wax, to
indicate the presence of adulterants such as paraffi
ns
 Iodine value
• Also known as Iodine number

• The number of grams of iodine absorbed

under specified conditions by 100g of oil, fats,
wax, or other substance.

• Quantitative measure of unsaturated fats

• It serves to characterize and indicate the

substances purity or if they contain admixtures.
• METHOD I: HANUS Method – Iodine monobromide
• METHOD II: WIJS Method – Iodine monochloride
IV = N x ( Vb- Va) x 0.1269 x 100
Wt in g
Expressed as gof iodine consumedby 100g
of oil
Classification:
Drying oil: IV> 120 Ex: Linseed Oil, Fish Oil
Semi-drying oil: IV of 100-120 Ex: Olive oil, Almond
oil Non-drying oil: IV < 100 Ex: Cotton seed oil,
Sesame Oil
 Hydroxyl value
• Hydroxyl number
• Is the number of mg of KOH equivalent to
the hydroxyl content of 1g of the substance.

• This constant gives an indication of the identity

and purity of fatty substance possessing
Hydroxyl groups.
 Acetyl value
• Number of milligrams of KOH required to
neutralize the acetic acid obtained by the
saponification of 1 g of acetyl ated fatty acids

A= S-F _ 1-(0.00075S)

Where:S =saponification value of acetyl ated fatty

acids
A =acetyl value of free fatty acids
F = acid value of original fatty acids
0.00075 = # of grams of acetyl group
corresponding to 1mg of KOH
 Volatile oils
• Also known as ethereal oils or essential oils.

• They are mixtures of hydrocarbon

terpenes, sesquiterpenes and polyterpenes
and their
oxygenated derivatives obtained from various
parts of the plant.

• Volatile oils evaporate on exposure to air

at ordinary temperature and are the
odorous constituents.
 Chemical components of volatile
oils
1. Hydrocarbons
2. Alcohols
3. Aldehydes
4. Ketones
5. Phenol s
6. Acids
7. Sulfuric compounds
• Analysis of volatile oils determines the
purity and value based on certain
physical characteristics or properties.
1. Odor
2 . Appearance and color

1. Specific gravity
2. Rotatory power - A substance’s ability to rotate the plane
of polarization of polarized electromagnetic radiation,
usually visible light.
3. Refractive index - measure of the bending of a ray of light
when passing from one mediu m into another.
4. Solidifying points
5. Solubility
6. Behavior during distillation
 Congealing point
• The highest temperature at which it solid ifies.

• The congealing point of the liquid is the same

as the melting temperature of the solid, but
since
the liquid may be cooled to a temperature below
its congealing point without assuming the solid
form, the method described is used to
determine the congealing point of a liquid or of a
melted
solid.
 Solubility of Essential oils
Generally most volatile oils are soluble
to organic solvents:
• Alcohol
• Ether
• Chloroform
• hexane
• Carbon disulfide
• Benzene
 Specific assays for Volatile oils
• Assay of
Alcohol content

• Assay of Phenol content

• Assay of Ester content

 Alkaloids
• Are obtained from plant, animal or
synthetic sources.

• Contains organic nitrogen within their

chemical structures that are usually
distinguished as
primary, secondary or tertiary.

• Possess physiological activity.

Assay for Alkaloids and preparations
are utilized for:
1. Standardization
2. Determination of purity
3. Commercial evaluation
4. P harmacological evaluation
Methods used to determine the
presence of alkaloids quantitatively:
1. Gravimetric
2. Volumetric
3. Spectrometric
4. Electrometric
5. Physiological
The amount of alkaloids that exists may
be subjected to variations in different
samples of the same drug:
1. Age of plant when collected
2. Season of the year
3. Type of soil and climate
4 . Conditions when the raw material is collected
Quality of alkaloids prese nt in Galenical
preparations is also subject to variat ions:
1. Quality of drug employed
2. Menstruum used in the extract of the alkaloid
3. Amount of decomposition of the alkaloid
during the process of extraction and storage
Factors that contributes to the
deterioration of alkaloid al preparations:
1. Nature of the alkaloid
2. pH value of the preparation
3. Heat
4. Light
 Physical Properties of Alkaloid
Solubility
• Alkaloids (R3N) or free alkaloids:
– Sparingly soluble in water
– Soluble in most organic solvents

• Alkaloid al salts
– Soluble in water
– Sparingly soluble in immiscible solvents
Chemical Properties of Alkaloids
• Reaction to acid
Alkaloids combine directly with acids will form salts that
are soluble in water but insoluble in certain
solvents
organic
R3N + HCl= R3NH Cl (water soluble salt)
• Reaction to base
Alkaloids will be liberated from aqueous solutions when
combine directly with alkalis
R3NH Cl + NaOH = R3N + NaCl + H2O
• Alkaloids form highly insoluble precipitates with reagents
that contain salts of heavy metals.
 Sources of errors in
Assaying Alkaloids
1. Failure to secure complete extraction of
the alkaloids from solution.
2. Loss of volatile solvent during maceration
of the drug before an aliquot portion is taken.
3. Imperfect separation of immiscible liquids.
4. Failure to wash properly the apparatus
being used for the assay
5. Decomposition of alkaloids
6. Use of wrong indicators.
Mercuric Iodide TS – Valser’s Reagent
 Preparation of drugs for assay
• The drug should be pulverized to
the required fineness
• Care should be taken to avoid the loss
of water during the pulverization

Weighing for assay: The accuracy should

be within 10mg for quantities equal to 5g
or
over
 Extraction
The alkaloid-bearing drugs are
extracted with one of the following
methods:
• Maceration
• Percolation
• Continuous Extraction
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