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A Seminar as a part of Curricular requirement for
M.Pharmacy I year II Semester
Presented by
P. Pavan kalyan
Reg.No.20L81S0705
Pharmaceutical Analysis
STUDY OF QUALITY OF RAW MTERIALS AND
GENERAL METHODS OF ANALYSIS OF RAW
MATERIALS USED IN COSMETIC MANUFACTURE
AS PER BIS
Contents:
• Types of raw materials used in used in cosmetics manufacture.
• General methods of analysis of raw materials used in cosmetic
manufacture as per BIS.
• References.
Types of raw materials used in cosmetic
manufacture
 Water
 Preservatives
 Humectants
 Surfactants
 Oils, fats & waxes
 Perfumes
 Colours
Water
Most widely used raw materials in the manufacture of cosmetics.
Water is used as a solvent for many ingredients of cosmetics. Water contains
inorganic ions like calcium, magnesium, sodium, Potassium, bicarbonate, sulphate,
chloride, silicates, etc.
• Presence of inorganic ions magnesium and zinc may interfere with static
charges of surfactants and it separates the emulsion.
• Presence of micro organisms if micro organism are present in the formulation of
cosmetics then leads to foul odour, visible colonies of bacteria, moulds, or fungi.
This may harm the cosmetics
Purification of water:
Purification of water ion exchange system, distillation, reverse osmosis,
microbiological purification
1. Chemical treatment:- HCHO, Sodium hypochlorite
2. Heat treatment:- water heated to 120 c
3. UV radiation:- below 300 nm has lethal effect on micro organism
4. Filtration:- pore size 0.2 milli micron or less to retain bacteria
General methods of analysis of raw materials
used in cosmetic manufacture as per BIS
1. Appearance (solid/liquid, flakes, granules, powdery, lumps)
2. Colour
3. pH ( Ph meter)
4. Melting point (sometimes melting range)
5. Relative density
6. Refractive index
7. bulk density
8. Loss on drying
9. loss on ignition/ residue on ignition
10. Total solids
11. Determination of Acid value
12. Determination of saponification value
13. Determination of iodine value
14. Determination of ester value
15. Determination of total ash
16. Matter insoluble in acid/ acid insoluble ash and water soluble ash
17. Sulphated ash.
18. Determination of moisture and volatile content.
19. Test for Arsenic and heavy metals
20. Test for Iron.
21. Test for free acid/alkali.
22. Microbial analysis.
23. Other tests for specific contents.
Melting point:
 Melt the material by warming it in a water bath at a temperature just sufficient
to melt it. Dip the thermometer and withdraw, so as to get the bulb thinly coated
with wax.
 Let it stand for 24hrs.
 Insert the thermometer into the test tube through the bored cork and then place
into the test-tube in the water bath .
 Raise the temp gradually at the rate of 1c in 3 mins.
 Note the temp accurately to 0.1c , at which a transparent drop from one end of
the thermometer bulb. Record this temp as melting point temp of the material.
Relative density:
 The relative density of glycerin shall be expressed as the ratio of the mass in
air of a given volume of material at 30c to that of an equal volume of water
at the same temp.
 Determined by hydrostatic balance or relative density bottle.
 A relative density bottle method of 25 or 50 ml capacity with central neck
filled with ground-glass thermometer which serves as a stopper, graduated
from 10 to 40c in 0.2c divisions and an overflow side tube with ground in
glass cap as shown below .
First we calibrate , weigh and then fill freshly with cooled
water at about 25c after removing the cap of the side arm.
Filling should be done until it overflows.
This test is done for most substances mainly liquid- raw
materials like glycerol, PEG, glycerine, sorbitol , iospropyl
alcohol etc
Relative density at 25c = A-B/C-B
A = mass of the relative density bottle with the material in g
B= mass of the bottle in g
C = mass of the bottle with water
Refractive index:
 Refractive index is a material constant, dependent on the chemical composition
of a substance.
 The instrument used is called as refractometer.
 Tungsten lamp is used as a monochromatic source with appropriate filters.
Sample is applied in between two prisms , closed and estimated.
 Its is test for many liquids ,semisolids and solids.
 Example: lanolin, parabens, glycerine, sorbitol etc
Bulk density:
 Weigh a 42g of fixed amount of powder , seived and then tranfered to
the cylinder of the bulk density apparatus.
 The apparatus lifts the cylinder to a heights of 25mm without jerks .
Continues lifting and dropping for 50 cycles
 Read the final volume.
 Calculation: bulkdensity = 40/v
 V= volume in ml of the sieved material.
Loss on drying :
 Weigh accurately about 5g of the material in a preweighed, clean and dry
squat form weighing bottle, and dry to constant mass at 105c ±2 °c
Calculation:
Loss on drying = 100 x W1/W2
W1= Loss in mass on drying
W2= Mass in g of the material taken for the test
 This is done for most of the raw materials like calcium carbonate , zinc oxide,
Mg carbonate and many others
Residue on ignition:
 Weigh accurately about 3g of the material in a tarred platinum or silica
dish. Slowly burn the material and finally ignite it in a furnace at 600 ±
25°c until all carbonaceous matter has disappeared. Cool in a desiccator
and weigh. Repeat till constant mass is obtained.
 Residue on ignition, (%mass) = 100 x (M1 – M2)/ M.
M1 = mass in g of the ignited residue with the crucible
M2 = mass in g of the crucible
M = mass in g of the material taken for the test.
 This is performed for isopropyl myristate some humectant raw materials
Loss on ignition:
 5g of sample ignited at 800°c to constant mass within 1mg.
 this is done for talc , titanium dioxide like substances.
Loss on ignition (%mass) = 100 x (M1-M2)/M1
M1 = Mass in g of the dried material taken for the test
M2= Mass in g of the residue.
Total solids:
 Water is removed by keeping the material vacuum at 80°c and the residue is
weighed.
 Weigh accurately about 1g of the sample in a suitable tarred weighing bottle ,
keep at a temperature of 80c and at pressure NMT 5mm of hg for 6 hrs.
transfer it into a desicator cool and weigh.
Total solids (%mass) = 100 x M1/M2
M1= Mass of the residue in the weighing bottle in g
M2 = mass of the material taken for the test.
Determination of acid value:
As the number of mg of alkali required to neutralize the free fatty acids present
in oil or fat substance
Used to determine the freshness of an oil / check the rancidity.
Acid value is an analytical tool used to determine the freshness of an oil. It
works on simple acid base titration method, the free fatty acids which are present
in an oil due to oxidative or hydrolytic process is titrated against a standard alkali
using phenolphthalein as an indicator. The amount of alkali consumed represents
the amount of fatty acid
Calculate the acid value:
Result = (Mol. Wt. X V) X (N/W)
Mol. Wt = molecular weight of potassium hydroxide, 56.11 V = volume (ml)
N = normality of the potassium hydroxide solution or the sodium hydroxide
solution
W = weight of the sample taken (g)
 Transfer a accurately weighed quantity 10 g of substance
 Into 250 ml conical flask
 50 ml of a mixture of equal volumes of alcohol and ether (previously
neutralised)
 If the test specimen does not dissolve in the cold solvent, connect the flask
with a suitable condenser and warm slowly, with frequent shaking, until the
specimen dissolves
 Add 1 ml of phenolphthalein and titrate with 0.1 N potassium hydroxide or
0.1 N sodium hydroxide until the solution remains faintly pink after shaking
for 30 s.
ACID VALUE SAMPLE WEIGHT (g)
0-1 20
1-4 10
4-15 2.5
15-74.9 0.5
≥75.0 0.1
Saponification value:
 The Saponification Value is the number of mg of potassium hydroxide required
to neutralize the free acids and saponify the esters contained in 1.0 g of the
substance
 Transfer a accurately weighed quantity 1.5-2 g of substance
 Into 250 ml conical flask
 Add to it 25.0 ml of 0.5 N alcoholic potassium hydroxide
 Heat the flask on a steam bath, under a suitable condenser to maintain reflux for
30 min, frequently rotating the contents
[NOTE: Reflux time can be up to 90 min to ensure complete saponification,
depending on the type of ester to be tested.]
Then add 1 ml of phenolphthalein and titrate the excess KOH
with 0.5 N hydrochloric acid .
Perform a blank determination.
Calculate the Saponification Value:
Result = [M x ( Vb – Vt ) X N] / W
M = molecular weight of potassium hydroxide, 56.11
Vb = volume of 0.5 N hydrochloric acid consumed in the blank test (ml)
Vt = volume of 0.5 N hydrochloric acid consumed in the actual test (ml)
N = exact normality of the hydrochloric acid
W = Weight of the substance taken for the test(g)
Iodine value:
The iodine value represents the number of g of iodine absorbed, under the
prescribed conditions, by 100 g of the substance. Unless otherwise specified in the
individual monograph, determine the iodine value by method (hanus method).
transfer a accurately weighed quantity (reference to table 3)
Into glass stopered , 250 ml conical flask.
Dissolve it in 10 ml of chloroform, add 25.0 ml of iodobromide.
Insert the stopper & allow it to stand for 30 min protected from light, with
occasional shaking.
 then add, in the order named, 30 ml of potassium iodide, 100 ml of water
 titrate the liberated iodine with 0.1 n sodium thiosulfate with shaking
thoroughly after each addition of thiosulfate.
When the iodine colour becomes quite pale add 3 ml of starch and continue
the titration until the blue colour is discharged.
 Perform a blank test.
Calculate the Iodine Value:
Result = [A x (Vb - Vs) X N]/(10 ´ W)
A = atomic weight of iodine, 126.90
Vb = volume of 0.1 N sodium thiosulfate consumed by the blank test (ml)
Vs = volume of 0.1 N sodium thiosulfate consumed by the actual test (ml)
N = exact normality of the sodium thiosulfate
W = weight of the substance taken for the test (g)
NOTE: If more than half of the iodobromide is absorbed by the portion of
the substance taken, repeat the determination, using a smaller portion of the
substance under examination.
Other tests performed similarly are hydroxyl value, ester value, peroxide
value.
These are usually performed for waxes, oils, fats etc.
IODINE VALUE RANGE SAMPLE WEIGHT(g)
<5 3.0
5-10 1.0
21-50 0.4
51-100 0.2
101-150 0.13
151-200 0.1
Ester value:
The Ester Value is the number of mg of potassium hydroxide required to
saponify the esters in 1.0 g of the substance. If the saponification value and the
acid value have been determined, the difference between these two represents
the Ester Value .
Ester Value = Saponification Value - Acid Value
Procedure:
Place 1.5-2 g of accurately weighed substance, into a 250 ml flask, add 20-30
ml of neutralized alcohol and shake. Add 1 ml of phenolphthalein and titrate
with 0.5 N alcoholic potassium hydroxide until the free acid is neutralized.
Add 25.0 ml of 0.5 N alcoholic potassium hydroxide and proceed as
directed in saponification value, beginning with “Heat the flask” and omitting the
further addition of phenolphthalein and titrate with standardised HCl acid,
calculate the Ester Value.
Result = [Mol wt X (Vb - Vt) x N] / W
Mol wt = molecular weight of potassium hydroxide, 56.11 VB = volume of 0.5 N
hydrochloric acid consumed in the blank test (ml)
Vt = volume of 0.5 N hydrochloric acid consumed in the actual test (ml)
N = exact normality of the hydrochloric acid
W = weight of the substance taken for the test (g)
Determination of moisture and volatile matter:
About 5gm of each sample material was weighed accurately and placed in a
porcelain or glass dish, about 6-8cm in diameter and about 2- 4cm in depth. It was
dried in an air oven at a temperature of 105°C to a constant mass.
Calculation:
The moisture and volatile matter of all samples were calculated by using the
following formula.
Moisture and volatile matter =100 M1 / M
M1 - loss of mass in g on drying and, M - Mass in grams of the material taken for
the test.
Determination of Ash Values
Total ash
Take about 2 or 3 g, accurately weighed, of the ground drug in a tarred platinum
or silica dish previously ignited and weighed. Scatter the ground drug in a fine
even layer on the bottom of the dish. Incarnated by gradually increasing the heat-
not exceeding dull red heat- until free from carbon, cool and weigh.
If a carbon free ash cannot be obtained in this way, exhaust the charred mass
with hot water, collect the residue on an ash less filter paper, increate the residue
and filter paper, add the filtrate, evaporate to dryness and ignite at low
temperature. Calculate the percentage of ash with reference to the air dried drug.
Acid- insoluble ash
Boil the total ash with for five minutes with 25 ml of dilute
hydrochloric acid, collect the insoluble matter in a Gooch crucible or on an ash
less filter paper, wash with hot water, ignite, and weigh. Calculate the
percentage of acid- insoluble ash with reference to the air dried drug.
Water- soluble ash
Boil the total ash for 5 minutes with 25 ml of water; collect the insoluble
matter in a Gooch crucible or on an ash less filter paper, wash with hot water,
and ignite to constant weight at a low temperature. Subtract the weight of
insoluble matter from the weight of the ash; the difference in weight represents
the water- soluble ash. Calculate the percentage of water- soluble ash with
reference to the air dried drug.
Sulphated ash
A silica crucible was heated to redness for 10 minutes, allowed to cool
in desiccators and weighed. 1 g of substance was accurately weighed and transferred
to the crucible. It was ignited gently at first, until the substance was thoroughly
charred. Then the residue was cooled and moistened with 1 ml concentrated sulfuric
acid, heated gently until white fumes are no longer evolved and ignited at 800° ±
25°C until all black particles have disappeared.
The ignition was conducted in a place protected from air currents. The crucible
was allowed to cool, and a few drops of concentrated sulfuric acid were added and
heated. Ignited as before, allowed to cool, and weighed. The operation was repeated
until two successive weighing does not differ by more than 0.5 mg. calculate the
percentage of Sulphated ash with reference to the air dried drug.
Determination of arsenic:
 Arsenic present in a solution of the material is reduced to arsine, which is made to
react with mercuric bromide paper.
 The stain produced is compared with a standard stain.
Procedure:
 Weigh 1g of the material in a platinum dish and add 5ml of conc sulphuric acid and
5 ml of conc nitric acid. Take to fumes on a hotplate.
 Cool and again take to fumes with three successive 5ml portions of HCl . Cool and
dissolve cautiously in water and makeup the volume exactly to 100 ml.
November 21 33
Carry out the test as prescribed in IS 2088, adding into the Gutzeit
bottle, 2 ml of Ferric ammonium sulphate solution, 0.5 ml of stannous
chloride solution and 25 ml of sample solution as prepared in F-4.
 For comparison, prepare a stain using 0.00 I mg of arsenic trioxide
Test for free acid or alkali:
Shake 20g of the material with an equal amount of hot distilled water. test the
aqueous portion with blue litmus and red litmus.
The material shall be taken to have passed the test if neither blue or red litmus
changes color.
This is done for glycerin, lanolin, ethyl parabes, methyl paraben, p-phenylene
diamine ,sorbitol etc..
Microbial tests:
The basic microbial analytical techniques include staining , grams
staining other test like tests for oxidase ,catalase etc.
Methods for Detection of Specific Organisms:
Principle :
The first step of the procedure is to perform enrichment by using a
nonselective broth medium to increase the number of micro-organisms
without the risk of inhibition by the selective ingredients that are present in
selective/differential growth media. The second step of the test (isolation) is
performed on a selective medium followed by identification tests.
Water-miscible products:
 Transfer the sample of product to a suitable container containing an
appropriate volume of broth.
Preparation of the initial suspension in the enrichment broth:
General method:
The enrichment is prepared from a sample of at least 1 g or 1 ml of the well-
mixed product under test, which is dispersed in at least 9 ml of enrichment
broth.
Water-immiscible products :
Transfer the sample of product to a suitable container containing a suitable
quantity of solubilizing agent (e.g. Polysorbate 80). Disperse the sample within the
solubilizing agent and add an appropriate volume of broth.
 In case if a common/ universal neutralizing broth is used, having solubilizing
agents, then bothh water miscible and immiscible products are to be treated in the
same manner as broth will contain emulsifying agents.
TYPE OF TEST TYPE OF RAW MATERIALS
1.Appearance
2.Colour
3.pH All raw materials
4.Melting point
5.Relative density
6.Refractive index
7.Bulk density All solid raw materials
8.Loss on drying Powdered material mainly
9.Loss on ignition
10.Total solids Glycerol, PEG, glycerine,
isopropyl alcohol,sorbital,lanoline
TYPE OF TEST TYPE OF RAW MATERIALS
11.Determination of Acid value
12. Determination of Saponication
value
13. Determination of iodine value
14. Determination of ester value
Oils, fats, waxes,
Eg: bees wax, paraffin wax, lanoline,
Stearic acid, oleic acid, coconut oil,
Seasame oil etc..
15. Determination of total ash
16.Matter insoluble in acid /acid
insoluble ash and water soluble ash
17.Sulphated ash
Glycerine , talc , titanium dioxide,
mg carbonate
18.Determination of moisture and
volatile content
Volatile solvents, other solvents likes
Acetone ,ethanol
19.Test for arsenic and heavy metals
20.Test for iron
21.Test for free acid/alkali
Mg carbonate , ca carbonate , cetyl
stearate , SLS etc…..
22.Microbial analysis All raw materials
Reference
 Bureau of Indian standards for cosmetic raw materials.
Study of Quality of Raw Materials and General methods of analysis of Raw materials used in cosmetic manufacture as per BSI

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Study of Quality of Raw Materials and General methods of analysis of Raw materials used in cosmetic manufacture as per BSI

  • 1. A Seminar as a part of Curricular requirement for M.Pharmacy I year II Semester Presented by P. Pavan kalyan Reg.No.20L81S0705 Pharmaceutical Analysis STUDY OF QUALITY OF RAW MTERIALS AND GENERAL METHODS OF ANALYSIS OF RAW MATERIALS USED IN COSMETIC MANUFACTURE AS PER BIS
  • 2. Contents: • Types of raw materials used in used in cosmetics manufacture. • General methods of analysis of raw materials used in cosmetic manufacture as per BIS. • References.
  • 3. Types of raw materials used in cosmetic manufacture  Water  Preservatives  Humectants  Surfactants  Oils, fats & waxes  Perfumes  Colours
  • 4. Water Most widely used raw materials in the manufacture of cosmetics. Water is used as a solvent for many ingredients of cosmetics. Water contains inorganic ions like calcium, magnesium, sodium, Potassium, bicarbonate, sulphate, chloride, silicates, etc. • Presence of inorganic ions magnesium and zinc may interfere with static charges of surfactants and it separates the emulsion. • Presence of micro organisms if micro organism are present in the formulation of cosmetics then leads to foul odour, visible colonies of bacteria, moulds, or fungi. This may harm the cosmetics
  • 5. Purification of water: Purification of water ion exchange system, distillation, reverse osmosis, microbiological purification 1. Chemical treatment:- HCHO, Sodium hypochlorite 2. Heat treatment:- water heated to 120 c 3. UV radiation:- below 300 nm has lethal effect on micro organism 4. Filtration:- pore size 0.2 milli micron or less to retain bacteria
  • 6. General methods of analysis of raw materials used in cosmetic manufacture as per BIS 1. Appearance (solid/liquid, flakes, granules, powdery, lumps) 2. Colour 3. pH ( Ph meter) 4. Melting point (sometimes melting range) 5. Relative density 6. Refractive index 7. bulk density 8. Loss on drying 9. loss on ignition/ residue on ignition
  • 7. 10. Total solids 11. Determination of Acid value 12. Determination of saponification value 13. Determination of iodine value 14. Determination of ester value 15. Determination of total ash 16. Matter insoluble in acid/ acid insoluble ash and water soluble ash 17. Sulphated ash. 18. Determination of moisture and volatile content. 19. Test for Arsenic and heavy metals 20. Test for Iron. 21. Test for free acid/alkali. 22. Microbial analysis. 23. Other tests for specific contents.
  • 8. Melting point:  Melt the material by warming it in a water bath at a temperature just sufficient to melt it. Dip the thermometer and withdraw, so as to get the bulb thinly coated with wax.  Let it stand for 24hrs.  Insert the thermometer into the test tube through the bored cork and then place into the test-tube in the water bath .  Raise the temp gradually at the rate of 1c in 3 mins.  Note the temp accurately to 0.1c , at which a transparent drop from one end of the thermometer bulb. Record this temp as melting point temp of the material.
  • 9. Relative density:  The relative density of glycerin shall be expressed as the ratio of the mass in air of a given volume of material at 30c to that of an equal volume of water at the same temp.  Determined by hydrostatic balance or relative density bottle.  A relative density bottle method of 25 or 50 ml capacity with central neck filled with ground-glass thermometer which serves as a stopper, graduated from 10 to 40c in 0.2c divisions and an overflow side tube with ground in glass cap as shown below .
  • 10. First we calibrate , weigh and then fill freshly with cooled water at about 25c after removing the cap of the side arm. Filling should be done until it overflows. This test is done for most substances mainly liquid- raw materials like glycerol, PEG, glycerine, sorbitol , iospropyl alcohol etc Relative density at 25c = A-B/C-B A = mass of the relative density bottle with the material in g B= mass of the bottle in g C = mass of the bottle with water
  • 11. Refractive index:  Refractive index is a material constant, dependent on the chemical composition of a substance.  The instrument used is called as refractometer.  Tungsten lamp is used as a monochromatic source with appropriate filters. Sample is applied in between two prisms , closed and estimated.  Its is test for many liquids ,semisolids and solids.  Example: lanolin, parabens, glycerine, sorbitol etc
  • 12. Bulk density:  Weigh a 42g of fixed amount of powder , seived and then tranfered to the cylinder of the bulk density apparatus.  The apparatus lifts the cylinder to a heights of 25mm without jerks . Continues lifting and dropping for 50 cycles  Read the final volume.  Calculation: bulkdensity = 40/v  V= volume in ml of the sieved material.
  • 13. Loss on drying :  Weigh accurately about 5g of the material in a preweighed, clean and dry squat form weighing bottle, and dry to constant mass at 105c ±2 °c Calculation: Loss on drying = 100 x W1/W2 W1= Loss in mass on drying W2= Mass in g of the material taken for the test  This is done for most of the raw materials like calcium carbonate , zinc oxide, Mg carbonate and many others
  • 14. Residue on ignition:  Weigh accurately about 3g of the material in a tarred platinum or silica dish. Slowly burn the material and finally ignite it in a furnace at 600 ± 25°c until all carbonaceous matter has disappeared. Cool in a desiccator and weigh. Repeat till constant mass is obtained.  Residue on ignition, (%mass) = 100 x (M1 – M2)/ M. M1 = mass in g of the ignited residue with the crucible M2 = mass in g of the crucible M = mass in g of the material taken for the test.  This is performed for isopropyl myristate some humectant raw materials
  • 15. Loss on ignition:  5g of sample ignited at 800°c to constant mass within 1mg.  this is done for talc , titanium dioxide like substances. Loss on ignition (%mass) = 100 x (M1-M2)/M1 M1 = Mass in g of the dried material taken for the test M2= Mass in g of the residue.
  • 16. Total solids:  Water is removed by keeping the material vacuum at 80°c and the residue is weighed.  Weigh accurately about 1g of the sample in a suitable tarred weighing bottle , keep at a temperature of 80c and at pressure NMT 5mm of hg for 6 hrs. transfer it into a desicator cool and weigh. Total solids (%mass) = 100 x M1/M2 M1= Mass of the residue in the weighing bottle in g M2 = mass of the material taken for the test.
  • 17. Determination of acid value: As the number of mg of alkali required to neutralize the free fatty acids present in oil or fat substance Used to determine the freshness of an oil / check the rancidity. Acid value is an analytical tool used to determine the freshness of an oil. It works on simple acid base titration method, the free fatty acids which are present in an oil due to oxidative or hydrolytic process is titrated against a standard alkali using phenolphthalein as an indicator. The amount of alkali consumed represents the amount of fatty acid
  • 18. Calculate the acid value: Result = (Mol. Wt. X V) X (N/W) Mol. Wt = molecular weight of potassium hydroxide, 56.11 V = volume (ml) N = normality of the potassium hydroxide solution or the sodium hydroxide solution W = weight of the sample taken (g)
  • 19.  Transfer a accurately weighed quantity 10 g of substance  Into 250 ml conical flask  50 ml of a mixture of equal volumes of alcohol and ether (previously neutralised)  If the test specimen does not dissolve in the cold solvent, connect the flask with a suitable condenser and warm slowly, with frequent shaking, until the specimen dissolves  Add 1 ml of phenolphthalein and titrate with 0.1 N potassium hydroxide or 0.1 N sodium hydroxide until the solution remains faintly pink after shaking for 30 s.
  • 20. ACID VALUE SAMPLE WEIGHT (g) 0-1 20 1-4 10 4-15 2.5 15-74.9 0.5 ≥75.0 0.1
  • 21. Saponification value:  The Saponification Value is the number of mg of potassium hydroxide required to neutralize the free acids and saponify the esters contained in 1.0 g of the substance  Transfer a accurately weighed quantity 1.5-2 g of substance  Into 250 ml conical flask  Add to it 25.0 ml of 0.5 N alcoholic potassium hydroxide  Heat the flask on a steam bath, under a suitable condenser to maintain reflux for 30 min, frequently rotating the contents [NOTE: Reflux time can be up to 90 min to ensure complete saponification, depending on the type of ester to be tested.]
  • 22. Then add 1 ml of phenolphthalein and titrate the excess KOH with 0.5 N hydrochloric acid . Perform a blank determination. Calculate the Saponification Value: Result = [M x ( Vb – Vt ) X N] / W M = molecular weight of potassium hydroxide, 56.11 Vb = volume of 0.5 N hydrochloric acid consumed in the blank test (ml) Vt = volume of 0.5 N hydrochloric acid consumed in the actual test (ml) N = exact normality of the hydrochloric acid W = Weight of the substance taken for the test(g)
  • 23. Iodine value: The iodine value represents the number of g of iodine absorbed, under the prescribed conditions, by 100 g of the substance. Unless otherwise specified in the individual monograph, determine the iodine value by method (hanus method). transfer a accurately weighed quantity (reference to table 3) Into glass stopered , 250 ml conical flask. Dissolve it in 10 ml of chloroform, add 25.0 ml of iodobromide. Insert the stopper & allow it to stand for 30 min protected from light, with occasional shaking.
  • 24.  then add, in the order named, 30 ml of potassium iodide, 100 ml of water  titrate the liberated iodine with 0.1 n sodium thiosulfate with shaking thoroughly after each addition of thiosulfate. When the iodine colour becomes quite pale add 3 ml of starch and continue the titration until the blue colour is discharged.  Perform a blank test.
  • 25. Calculate the Iodine Value: Result = [A x (Vb - Vs) X N]/(10 ´ W) A = atomic weight of iodine, 126.90 Vb = volume of 0.1 N sodium thiosulfate consumed by the blank test (ml) Vs = volume of 0.1 N sodium thiosulfate consumed by the actual test (ml) N = exact normality of the sodium thiosulfate W = weight of the substance taken for the test (g) NOTE: If more than half of the iodobromide is absorbed by the portion of the substance taken, repeat the determination, using a smaller portion of the substance under examination.
  • 26. Other tests performed similarly are hydroxyl value, ester value, peroxide value. These are usually performed for waxes, oils, fats etc. IODINE VALUE RANGE SAMPLE WEIGHT(g) <5 3.0 5-10 1.0 21-50 0.4 51-100 0.2 101-150 0.13 151-200 0.1
  • 27. Ester value: The Ester Value is the number of mg of potassium hydroxide required to saponify the esters in 1.0 g of the substance. If the saponification value and the acid value have been determined, the difference between these two represents the Ester Value . Ester Value = Saponification Value - Acid Value Procedure: Place 1.5-2 g of accurately weighed substance, into a 250 ml flask, add 20-30 ml of neutralized alcohol and shake. Add 1 ml of phenolphthalein and titrate with 0.5 N alcoholic potassium hydroxide until the free acid is neutralized.
  • 28. Add 25.0 ml of 0.5 N alcoholic potassium hydroxide and proceed as directed in saponification value, beginning with “Heat the flask” and omitting the further addition of phenolphthalein and titrate with standardised HCl acid, calculate the Ester Value. Result = [Mol wt X (Vb - Vt) x N] / W Mol wt = molecular weight of potassium hydroxide, 56.11 VB = volume of 0.5 N hydrochloric acid consumed in the blank test (ml) Vt = volume of 0.5 N hydrochloric acid consumed in the actual test (ml) N = exact normality of the hydrochloric acid W = weight of the substance taken for the test (g)
  • 29. Determination of moisture and volatile matter: About 5gm of each sample material was weighed accurately and placed in a porcelain or glass dish, about 6-8cm in diameter and about 2- 4cm in depth. It was dried in an air oven at a temperature of 105°C to a constant mass. Calculation: The moisture and volatile matter of all samples were calculated by using the following formula. Moisture and volatile matter =100 M1 / M M1 - loss of mass in g on drying and, M - Mass in grams of the material taken for the test.
  • 30. Determination of Ash Values Total ash Take about 2 or 3 g, accurately weighed, of the ground drug in a tarred platinum or silica dish previously ignited and weighed. Scatter the ground drug in a fine even layer on the bottom of the dish. Incarnated by gradually increasing the heat- not exceeding dull red heat- until free from carbon, cool and weigh. If a carbon free ash cannot be obtained in this way, exhaust the charred mass with hot water, collect the residue on an ash less filter paper, increate the residue and filter paper, add the filtrate, evaporate to dryness and ignite at low temperature. Calculate the percentage of ash with reference to the air dried drug.
  • 31. Acid- insoluble ash Boil the total ash with for five minutes with 25 ml of dilute hydrochloric acid, collect the insoluble matter in a Gooch crucible or on an ash less filter paper, wash with hot water, ignite, and weigh. Calculate the percentage of acid- insoluble ash with reference to the air dried drug. Water- soluble ash Boil the total ash for 5 minutes with 25 ml of water; collect the insoluble matter in a Gooch crucible or on an ash less filter paper, wash with hot water, and ignite to constant weight at a low temperature. Subtract the weight of insoluble matter from the weight of the ash; the difference in weight represents the water- soluble ash. Calculate the percentage of water- soluble ash with reference to the air dried drug.
  • 32. Sulphated ash A silica crucible was heated to redness for 10 minutes, allowed to cool in desiccators and weighed. 1 g of substance was accurately weighed and transferred to the crucible. It was ignited gently at first, until the substance was thoroughly charred. Then the residue was cooled and moistened with 1 ml concentrated sulfuric acid, heated gently until white fumes are no longer evolved and ignited at 800° ± 25°C until all black particles have disappeared. The ignition was conducted in a place protected from air currents. The crucible was allowed to cool, and a few drops of concentrated sulfuric acid were added and heated. Ignited as before, allowed to cool, and weighed. The operation was repeated until two successive weighing does not differ by more than 0.5 mg. calculate the percentage of Sulphated ash with reference to the air dried drug.
  • 33. Determination of arsenic:  Arsenic present in a solution of the material is reduced to arsine, which is made to react with mercuric bromide paper.  The stain produced is compared with a standard stain. Procedure:  Weigh 1g of the material in a platinum dish and add 5ml of conc sulphuric acid and 5 ml of conc nitric acid. Take to fumes on a hotplate.  Cool and again take to fumes with three successive 5ml portions of HCl . Cool and dissolve cautiously in water and makeup the volume exactly to 100 ml. November 21 33
  • 34. Carry out the test as prescribed in IS 2088, adding into the Gutzeit bottle, 2 ml of Ferric ammonium sulphate solution, 0.5 ml of stannous chloride solution and 25 ml of sample solution as prepared in F-4.  For comparison, prepare a stain using 0.00 I mg of arsenic trioxide Test for free acid or alkali: Shake 20g of the material with an equal amount of hot distilled water. test the aqueous portion with blue litmus and red litmus. The material shall be taken to have passed the test if neither blue or red litmus changes color. This is done for glycerin, lanolin, ethyl parabes, methyl paraben, p-phenylene diamine ,sorbitol etc..
  • 35. Microbial tests: The basic microbial analytical techniques include staining , grams staining other test like tests for oxidase ,catalase etc. Methods for Detection of Specific Organisms: Principle : The first step of the procedure is to perform enrichment by using a nonselective broth medium to increase the number of micro-organisms without the risk of inhibition by the selective ingredients that are present in selective/differential growth media. The second step of the test (isolation) is performed on a selective medium followed by identification tests.
  • 36. Water-miscible products:  Transfer the sample of product to a suitable container containing an appropriate volume of broth. Preparation of the initial suspension in the enrichment broth: General method: The enrichment is prepared from a sample of at least 1 g or 1 ml of the well- mixed product under test, which is dispersed in at least 9 ml of enrichment broth.
  • 37. Water-immiscible products : Transfer the sample of product to a suitable container containing a suitable quantity of solubilizing agent (e.g. Polysorbate 80). Disperse the sample within the solubilizing agent and add an appropriate volume of broth.  In case if a common/ universal neutralizing broth is used, having solubilizing agents, then bothh water miscible and immiscible products are to be treated in the same manner as broth will contain emulsifying agents.
  • 38. TYPE OF TEST TYPE OF RAW MATERIALS 1.Appearance 2.Colour 3.pH All raw materials 4.Melting point 5.Relative density 6.Refractive index 7.Bulk density All solid raw materials 8.Loss on drying Powdered material mainly 9.Loss on ignition 10.Total solids Glycerol, PEG, glycerine, isopropyl alcohol,sorbital,lanoline
  • 39. TYPE OF TEST TYPE OF RAW MATERIALS 11.Determination of Acid value 12. Determination of Saponication value 13. Determination of iodine value 14. Determination of ester value Oils, fats, waxes, Eg: bees wax, paraffin wax, lanoline, Stearic acid, oleic acid, coconut oil, Seasame oil etc.. 15. Determination of total ash 16.Matter insoluble in acid /acid insoluble ash and water soluble ash 17.Sulphated ash Glycerine , talc , titanium dioxide, mg carbonate 18.Determination of moisture and volatile content Volatile solvents, other solvents likes Acetone ,ethanol 19.Test for arsenic and heavy metals 20.Test for iron 21.Test for free acid/alkali Mg carbonate , ca carbonate , cetyl stearate , SLS etc….. 22.Microbial analysis All raw materials
  • 40. Reference  Bureau of Indian standards for cosmetic raw materials.