Weight/ml Moisture contents Loss on drying Toxicity tests Identification test Evaluation of ointments Ash test Alkalinity of glass

1. Dr. Asra Hameed
Pharm.D (JUW)
asra_hameed1@hotmail.com

2. Weight/ml
Moisture contents
Loss on drying
Toxicity tests
Identification test
Evaluation of ointments
Ash test
Alkalinity of glass

4. • “The weight per
milliliter of a liquid is
the weight expressed
in grams of 1 milliliter
of a liquid when
weighed in air at the
specific temperature”
Important
for
tinctures
and
extracts
and
defined as

5. Almost equal to density of the liquid,
For this,
Weigh a clean and dry
pycnometer accurately
Fill it with liquid, remove
any access and weigh
Determine the weight of liquid by
subtraction

6. Determine wt/ml by dividing
the weight of liquid that
filled the pycnometer by the
capacity of pycnometer
Expressed in g/ml

8. To measure the
amount of water and
volatile matters
Sample is dried
under specified
conditions.

9. Dry the sample at temperature specified in monograph, close the bottle. Cool
in desiccator & weigh
Place the loaded bottle in drying chamber, remove stopper and place it also in
the chamber.
Weigh and shake to evenly distribute the substance.
Take 1-2g of substance, reduce particle size if necessary, in a dried glass
stoppered, weighed bottle.

10. If “dry to constant weight”, until two
consecutive weighings do not differ
by more than 0.50 mg per g of
substance
Where capsules are to be tested,
use a portion of the mixed contents
of not fewer than 4 capsules.
Where tablets are to be tested, use
powder from not fewer than 4 tablets
USP <731> Loss On Drying

11. If the substance melts at a temperature at
which loss on drying is to be performed, it
is kept at 5 -10oC for 1-2 h for drying.
For substance requiring
thermogravimetric analysis a sensitive
electrobalance is used.
For vacuum drying vacuum desiccator or
vacuum drying pistol are used.
USP <731> Loss On Drying

13. Heating method/
thermogravimetry
• Volumetric method
• Coulometric method
Karl
Fischer

14. Accurately weighed powder (2 g) of the plant material is
taken in a tarred silica crucible.
The powder is the dried in an oven at 105 oC for 30 min,
cooled at room temperature in desiccator until constant
weight and weighed to calculate the moisture contents.
The results are expressed as a percent of dry powder.

15. Karl Fisher Method---Standard
technique for low moisture products.
Especially good for reducing sugars,
protein-rich food and foods and
pharmaceuticals with high volatile
oils.

16. 2 H2O + SO2 + I2 = H2SO4 + 2 HI
German
Petrochemist,
1901 – 1958
Publication:
1935

17.  KF Reaction
SO2 + RN + ROH ------> (RNH)SO3R
a sulfite compound
(RNH)SO3R + H2O + I2 + 2RN ------> (RNH)SO4R + 2(RNH)I
a sulfate compound
 Summary
H2O + I2 + SO2 + 3RN + ROH ----->(RNH)SO4R +
2(RNH)I
• The solvent (generally methanol) is involved in the reaction
• A suitable base keeps the pH 5 - 7

18. N NNNNNN NNNN
NN
H2O
CH3OH
H SO4CH3
I2 SO2 H I SO3
SO3
+ + + 2 +
+
Brown Mahogany Color
1.
2.

19. Karl Fisher Reagent: Dissolve Iodine in the
mixture of Pyridine, MeOH and SO2.
Titrate 100 mg of H2O with Carl Fisher Reagent.
Calculate Concentration of K F Reagent =mg
H2O /ml of Reagent
% H2O = Concentration x ml Reagent / mg of
Sample x 100

21. Usually performed on finished dosage
form of toxoids and plastic containers in
order to determine there safety or toxicity.
DIPHTHERIA TOXOID
Inject IV/SC 4 healthy guinea pigs (300-
400g) with a of DT that is atleast 5 times
the human immunizing dose (not less than
2ml).
Comply if no symptom of DT poisoning
appear within 30 days.

22. TETANUS TOXOID
Perform similar as DT but it
is given SC.
Check the symptoms of
tetanus toxin poisoning after
21 days.
PLASTIC CONTAINERS
Extract of plastic containers
used for parenteral
preparations is prepared.
Its IV injection to mice
determine its gross toxicity
and mortality.

23. Give IC into
dorsal
shaved skin
of rabbit or
The
standardized
plastic strips
are implanted
The local
reaction is
observed
for toxicity.

25. Place a
quantit
y of the
substa
nce
3 drops
of
phosph
oric
acid
a drop
of
lanthan
um
nitrate
a drop
of
iodine
a drop
of
ammon
ia
A
blue
colour
slowly
appears

26. substance
1 g of
magnesium
oxide
Alkaline
vapours
evolve
gradually
turn silver
paper
black

27. solution in
hydrochloric
acid
dilute 10
times with
water
white
precipitate
turns dark
brown on
the addition of
sodium
sulfide

28. solut
ion
acidify
with
sulfuric
acid
mix
with
chlor
ine
brown
solution
shaking
with
chlorofo
rm
Colourless
chloroform
layer turns
reddish.

29. 1 drop of a
solution
4 drops of
glyoxal
bis(2-
hydroxyanil)
1 drop of
sodium
hydroxide
A
reddish
brown
precipit
ate
dissolves in
chloroform
red
solution.

30. Substance
manganese
dioxide
sulfuric acid
evolved
chlorine
greenish
colour
blue
coloration
of
moistene
d starch

31. solution
sulfuric
acid
potassium
nitrite
brown
solution
results;
shaking
with
chloroform
colourless,
chloroform
layer turns
violet.

32. Solution
ferrous
sulfate
sulfuric
acid
brown
colour

33. alkaline solution
sodium
tetraphenylborate
white precipitate

34. solution
ferric
chloride
reddish
violet
colour
acetic
acid
disappe
ars on
HCl
white
crystalli
ne
precipit
ate

35. solution
acetic acid
uranyl/zinc
acetate
yellow
crystalline
precipitate

36. Spectrophotometric tests contribute meaningfully toward the identification
of many compendial chemical substances. The test procedures that follow
are applicable to substances that absorb IR and/or UV radiation

37. The IR absorption spectrum of a substance, compared with that
obtained concomitantly for the corresponding USP Reference
Standard, provides perhaps the most conclusive evidence of the
identity of the substance that can be realized from any single test.
The UV absorption spectrum, does not exhibit a high degree of
specificity.
Conformance with both IR absorption and UV absorption test
specifications, as called for in a large proportion of compendial
monographs, leaves little doubt, if any, regarding the identity of the
specimen under examination.

39. Chromatography is defined as a procedure by
which solutes are separated by a dynamic
differential migration process in a system
consisting of two or more phases, one of which
moves continuously in a given direction and in
which the individual substances exhibit different
mobilities by reason of differences in adsorption,
partition, solubility, vapor pressure, molecular
size, or ionic charge density. The individual
substances thus separated can be identified or
determined by analytical procedures.

40. The types of chromatography useful in qualitative and quantitative analysis that are
employed in the USP procedures are column, gas, paper, thin-layer, (including high-
performance thin-layer chromatography), and pressurized liquid chromatography
(commonly called high-pressure or high-performance liquid chromatography).
Paper and thin-layer chromatography are ordinarily more useful for purposes of
identification, because of their convenience and simplicity.
Column chromatography offers a wider choice of stationary phases and is useful for
the separation of individual compounds, in quantity, from mixtures.
Modern high-performance thin-layer chromatography, gas chromatography, and
pressurized liquid chromatography require more elaborate apparatus but usually
provide high resolution and identify and quantitate very small amounts of material.

41. Chromatographic identification by these methods
under given conditions strongly indicates identity
but does not constitute definitive identification.
Specific and pertinent chemical, spectroscopic, or
physicochemical identification of the eluted
component combined with chromatographic
identity is the most valid criterion of identification.

43. • Cracking of creams (separation
of oil and water)
• Development of granular and
lumpy appearance
• Marked change in viscosity
• Crystal growth
• Microbial contamination
The main
characteristics
need to be
checked are

44. Dilute a suitable qty of preparation with equal
volume of glycerol or liquid paraffin, as specified
Mount on a glass slide and examine under light
microscope
Count the number of particles with daimeter above
or below than that specified in monograph
Compare the percentage with official limits

45. Applies to those products in which labeled net
weight is not more than 150g
Select 10 filled containers, remove the label,
clean and weigh individually
Remove the contents by cutting the
containers and wash with suitable solvent

46. Dry and again weigh each empty container
together with its corresponding part, take
difference as weight of contents.
The average net weight of contents of 10
containers should not be less than the
labeled amount
The net weight of contents of any single
container should not be less than 90% of
the labeled amount (for ≤ 60g)

47. And not less than 95% of the labeled
amount (60-150g)
If this requirement is not met repeat this
procedures taking additional 20 containers
The average net weight of contents of 30
containers should not be less than labeled
amount

48. Contents of not more than 1 of the
30 units should be less than 90%
of the stated amount (for ≤ 60g)
And not less than 95% of the
labeled amount (60-150g)

49. The
preparation
should be
soluble in 9
parts of water
and 1.7 parts
of hot water
The
preparation
should be
miscible with
alcohol, ether
and
chloroform

50. Viscosity is determined using a method
specified in official monograph

51. Assay of active
ingredients
should be
performed
according to
monograph
Percentage
contents should
be within the
official limits

52. Microorganisms can grow, if no preservative is
added, or even if added, its efficiency is reduced
due to interaction with other ingredients
Microorganisms may get into the preparation during
handling and storage
Therefore, aseptic techniques of handling are
needed

53. • Direct inoculation
method
• Membrane
filtration method
Antimicrobial
assay
should be
performed
according to
official
monograph,
usually

54. Extrude completely the contents of 10 containers separately into flat bottom Petri dishes
Cover, heat at 85oC for 2 h and cool slowly to solidify
Remove the cover and invert on the stage of microscope adjusted to 30 times
magnification and equipped with eye-piece micrometer disk calibrated for this
magnification.
Additional illuminator may also used
Examine the entire bottom of Petri dish for metal particles.

55. Count the metal particles of 50µm or larger
The requirement is met if total number of such particles in all 10 tubes
does not exceed 50
Not more than 1 tube contains more than 8 such particles
If not, repeat with 20 more tubes
The requirement is met if total number of such particles in 30 tubes is
not more than 150
And not more than 3 containers allowed to contain more than 8
particles

57. Acid soluble ash
Acid insoluble ash
Water soluble ash
Sulphated ash

61. 2 gram
sample in
tarred
silica
crucible
moistened
with
H2SO4
ignited
gently
Crucible is
then cool
in
desiccator
again
moistened
with
H2SO4
reignited
and
weighed.
The
process is
repeated
till a
constant
weight is
obtained.
The
percentage
of
sulphated
ash with
reference
to air dried
sample
was
calculated

63. Alkalinity is a measure of
the ability of a solution to
neutralize acids to the
equivalence point of
carbonate or bicarbonate.

64. • Neutral glass
• High hydrolytic resistance
• Suitable for preparations whether or not for
parenteral use
TYPE 1
• Soda lime silica glass
• High hydrolytic resistance
• Suitable for acid,neutral & aqueous
preparations
TYPE 2
• Soda lime silica glass
• Moderate hydrolytic resistance
• Suitable for non-aqueous
preparations,powders for parenteral use
TYPE 3

66. In all glass, the sodium and potassium oxides
are hygroscopic.
The surface of the glass absorbs moisture
from the air.
The absorbed moisture and exposure to
carbon dioxide causes the NaO2 or NaOH and
KO2 or KOH to convert to sodium or
potassium carbonate.
Both NaCO2 and KCO2 are extremely
hygroscopic.

68. Take sufficient containers, not less than 3 from each batch,
so that the total volume of water to be tested is not less
than 250 ml. rinse the containers thoroughly with distilled
water and complete the rinsing with redistilled water.
Fill each container to 90% of its overflow capacity with the
redistilled water above. Cover the unsealed containers with
crimped pieces of new tin foil wash thoroughly with
acetone.
Place the containers on the rack in wash autoclave and
close the door securely, leaving the vent cock and open.

69. Heat until steam issues vigorously from the vent cock and
continue heating for 10 min. close the vent cock adjust the
heating so that the temperature rises 1 ċ/min until it reaches
121ċ, taking 20 to 25 min to reach that temperature. Keep the
temperature at 121ċ ± 0.5 ċ for one hour.
At the end of that period decrease the supply of heat and
cool at the rate of 0.5 ċ per min, until the internal pressure is
equal to the atmospheric pressure.
The time to cool from 121ċ – 100 ċ should be from 40 to 50
min.

70. open the autoclave take out the containers and
allow to cool them to 25 ċ, transfer 100 ml of water
from each container add 5 drops of methyl red
solution, and titrate with 0.01 N sulphuric acid.
The time elapsing between opening the autoclave
and titrating should not exceed 60 minutes. Carry
out blank test on 100 ml of water from the same
lot, and make the necessary correction.
The quantity of 0.01N sulphuric acid used for
containers with a capacity of up to 100 ml should
be, not more than 1.5 ml and for containers of
capacity greater than 100ml, not more than 0.5 ml.