A.

REFERENCE
National Formulary-27,2009 p. 1272-1273

B. PROCEDURE
1. DESCRIPTION
a. Material
Magnesium Stearate in-house standard

b. Procedure
Detemine form, color and odor of sample by visual or organoleptic examination.

2. SOLUBILITY
a. Material
Water
Alcohol
Ether
b. Procedure
For material and procedure see General Test Procedure Solubility, ANA-GT-027-xx.

3. IDENTIFICATION
Test A
a. Material
Diluted nitric acid (105mL/L)
Ether
ammonium chloride
Ammonium carbonate TS.
Dibasic sodium phosphate TS
6 N ammonium hydroxide
.
b. Procedure
Mix 5.0 g with 50 mL of peroxide-free ether, 20 mL of diluted nitric acid, and 20 mL of water in
a round-bottom flask
Connect the flask to a reflux condenser, and reflux until dissolution is complete. Allow to cool,
and transfer the contents of the flask to a separator.
Shake, allow the layers to separate, and transfer the aqueous layer to a flask. Extract the
ether layer with two 4-mL portions of water, and add these aqueous extracts to the main
aqueous extract. Wash the aqueous extract with 15 mL of peroxide-free ether.
Transfer the aqueous extract to a 50-mL volumetric flask, dilute with water to volume, and
mix. Retain this solution for the Limit of chloride and Limit of sulfate tests.
This solution responds to the test for Magnesium:
 - Solutions of magnesium salts in the presence of ammonium chloride yield no more than a
slightly hazy precipitate when neutralized with ammonium carbonate TS.
- But on the subsequent addition of dibasic sodium phosphate TS a white, crystalline
precipitate, which is insoluble in 6 N ammonium hydroxide, is formed.
Test B
a. Material
-
b. Procedure
The retention times of the peaks corresponding to stearic acid and palmitic acid in the
chromatogram of the Test solution correspond to those in the chromatogram of the System
suitability solution, as obtained in the Relative content of stearic acid and palmitic acid test.

4. MICROBIAL LIMIT
See Prosedur Tetap Uji Mikrobiologi (QC3-SOP-04/xx).
See Prosedur Tetap Pengujian Total Plate Count dan Kuman Pathogen (QC3-SOP-44/xx)

5. ACIDITY OR ALKALINITY
a. Material
Water
0.1 N hydrochloric acid
0.1 N sodium hydroxide
Bromothymol blue TS

b. Procedure
Transfer 1.0 g to a 100-mL beaker, add 20 mL of carbon dioxide-free water.
Boil on a steam bath for 1 minute with continuous shaking, cool.
Filter. Add 0.05 mL of bromothymol blue TS to 10 mL of the filtrate.
Add 0.1 N HCl or 0.1 N NaOH until the color of the solution is change.
Measure the 0.1 N HCl or 0.1 N NaOH needed.

6. LOSS ON DRYING
For material and procedure see General Test Procedure Loss on Drying, ANA-GT-004-x.
Condition test : Dry it at at 105oC to contant weight.

7. LIMIT OF CHLORIDE
a. Material
Nitric acid
Silver nitrate TS
0.02 N HCl
b. Procedure
 Transfer 10 mL of an aqueous solution obtained from Identification test A, into a Nessler tube
Add water to make 30-40 mL, and, if necessary, neutralize the solution with HNO 3 to litmus
Add 1 mL of HNO3, 1 mL of AgNO3 TS and water to make 50 mL. Mix and allow to stand for 5
minutes, protected from direct sunlight.
Compare the turbidity, if any, with 1.4 mL of 0.02 N HCl.

8. LIMIT OF SULFATE
a. Material
3 N Hydrochloric acid
Barium chloride TS
0.02 N Sulfuric acid

b. Procedure
Transfer 3 mL of an aqueous solution obtained from Identification test A, into a Nessler tube.
Add water to make 30-40 mL and if necessary neutralize the solution with HCl to litmus.
Add 1 mL of 3 N HCl, 3 mL of BaCl2 TS and water to make 50 mL.
Mix and allow to stand for 10 minutes.
Compare the turbidity, if any, with 3.0 mL of 0.02 N H 2SO4

9. LEAD
a. Materials
0.2 N nitric acid
Ammonia-cyanide solution
Dissolve 2 g Potassium cyanide in 15 mL of Ammonium Hydroxide, and dilute with water to 100 mL.
Ammonium citrate solution
Dissolve 40 g of citric acid in 90 mL of water. Add 2 or 3 drops of phenol red TS, then cautiously
add Ammonium Hydroxide until the solution acquires a reddish color. Remove any lead that may be
present by extracting the solution with 20-mL portion of dithizone extraction solution (see below),
until the dithizone solution retains its orange-green color.

Lead nitrate stock solution

Accurately weigh 159.8 mg of lead nitrate and quantitatively transfer into Add 1 mL nitric acid to 100
ml water. Dissolve 159.8 mg of lead nitrate in those. Dilute to 1000 mL with water. Prepare and
store this solution in glass container free from soluble lead salts.
Standard lead solution
On the day of use, accurately pipette 10 mL lead nitrate stock solution. Quantitatively transfer into
100-mL volumetric flask and dilute to volume with water, mix well.
Dithizone extraction solution
 Dissolve 30 mg of dithizone in 1000 mL of chloroform, and add 5 mL of alcohol. Store the solution in
refrigerator. Before use, shake a suitable volume of this solution with about half its volume of dilute
nitric acid (1 in 100), discarding the nitric acid.
Hydroxylamine hydrochloride solution
Dissolve 20 g of hydroxylamine hydrochloride in sufficient water to make approximately 65 mL.
Transfer to a separatory funel, add 5 drops of thymol blue TS, then add NH 4OH until the solution
assumes a yellow color. Add 10 mL of sodium diethyldithiocarbamate soluton (1 in 25), mix and
allow to stand for 5 minutes. Extract this solution with successive 10 to 15-mL portion of chloroform
until a 5-mL portion of the chloroform extract does not assume a yellow color when shaken with
cupric sulfate TS. Add 3 N HCl until the solution is pink (if necessary, add 1 or 2 drops more of
thymol blue TS), and then dilute with water to 100 mL.
Potassium cyanide solution
Dissolve 50 g of potassium cyanide to 100 mL with water. Remove any lead that may be present by
extracting the solution with 20-mL portion of dithizone extraction solution, until the dithizone solution
retains its orange-green color. Extract any dithizone remaining by shaking with chloroform. Dilute
the cyanide solution to 100 mL with water.
Standard dithizone solution
Dissolve 10 mg dithizone in 1000 mL of chloroform. Keep the solution in a glass-stoppered, lead
free bottle, suitable wrapped to protect it from light, and store in a refrigerator

b. Procedure
Standard preparation
To 20 mL of 0.2 N nitric acid add 5 g of lead, 4 mL of Ammonia-Cyanide Solution, and 2 drops of
Hydroxylamine Hydrochloride Solution, and shake with 10.0 mL of Standard Dithizone Solution for
30 seconds.
Test Preparation
Ignite 0.50 g in a silica crucible in a muffle furnace at 475 to 500 for 15 to 20 minutes. Cool, add
3 drops of nitric acid, evaporate over a low flame to dryness, and again ignite at 475 to 500 for
30 minutes.
Dissolve the residue in 1 mL of a mixture of equal parts by volume of nitric acid and water, and
wash into a separator with several successive portions of water.
Add 3 mL of Ammonium Citrate Solution and 0.5 mL of Hydroxylamine Hydrochloride Solution,
and render alkaline to phenol red TS with ammonium hydroxide. Add 10 mL of Potassium
Cyanide Solution.
Immediately extract the solution with successive 5-mL portions of Dithizone Extraction Solution,
draining off each extract into another separator, until the last portion of dithizone solution retains
its green color. Shake the combined extracts for 30 seconds with 20 mL of 0.2 N nitric acid, and
discard the chloroform layer.
Add to the acid solution 4.0 mL of Ammonia-Cyanide Solution and 2 drops of Hydroxylamine
Hydrochloride Solution.
Add 10.0 mL of Standard Dithizone Solution, and shake the mixture for 30 seconds. Pass the
chloroform layer through an acid-washed filter paper into a color-comparison tube.
Compare the color of chloroform layer of sample against standard
10.RELATIVE CONTENT OF STEARIC ACID AND PALMITIC ACID
a. Material
USP Stearic Acid RS
USP Palmitic Acid RS
Boron trifluoride
n-heptane
anhydrous sodium sulfate
Standard Solution
Transfer about 50 mg each of USP Stearic Acid RS and USP Palmitic Acid RS to a small conical flask
fitted with a suitable reflux condenser.
Add 5.0 mL of a solution prepared by dissolving 14 g of boron trifluoride in methanol to make 100 mL,
swirl to mix, and reflux for 10 minutes until the solids have dissolved. Add 4 mL of chromatographic n-
heptane through the condenser, and reflux for 10 minutes.
Cool, add 20 mL of saturated sodium chloride solution, shake, and allow the layers to separate. Pass
the n-heptane layer through 0.1 g of anhydrous sodium sulfate (previously washed with
chromatographic n-heptane) into a suitable flask.
Transfer 1.0 mL of this solution to a 10-mL volumetric flask, dilute with chromatographic n-heptane to
volume, and mix.
Test solution
Transfer about 100 mg of Magnesium Stearate, accurately weighed, to a small conical flask fitted with
a suitable reflux condenser, and proceed as directed for System suitability solution, beginning with
Add 5.0 mL of a solution prepared by dissolving.

Chromatographic system (see Chromatography 621 ) The gas chromatograph is equipped with
a flame-ionization detector, maintained at about 260; a splitless injection system; and a 0.32-mm
30-m fused silica capillary column bonded with a 0.5-m layer of phase G16. The column temperature
is maintained at 70 for about 2 minutes after injection, then programmed to increase at the rate of 5
per minute to 240 and to maintain this temperature for 5 minutes. The injection port temperature is
maintained at about 220. The carrier gas is helium with a linear velocity of about 50 cm per second.
Chromatograph the System suitability solution, and record the peak responses as directed for
Procedure: the relative retention times are about 0.86 for methyl palmitate and 1.0 for methyl stearate;
the resolution, R, between the methyl palmitate and methyl stearate peaks is not less than 5.0; the
relative standard deviation of the peak area responses for the palmitate and stearate peaks for
replicate injections is not greater than 6.0%; and the relative standard deviation of the peak area
response ratio of the palmitate to stearate peaks from these replicate injections is not more than 1.0%.
b. Procedure
Inject about 1 L of the Test solution into the chromatograph, record the chromatogram, and
measure the peak areas for all the fatty acid ester peaks in the chromatogram
Calculate the percentage of stearic acid in the fatty acid fraction of Magnesium Stearat
Formula : 100A/B
Where :
A = the area due to the methyl stearate peak
 B = the sum of the areas of all the fatty acid ester peaks in the chromatogram. calculate the
percentage of palmitic acid in the portion of Magnesium Stearate taken. The stearate peak
comprises not less than 40%; and the sum of the stearate and palmitate peaks is not less than
90% of the total area of all fatty acid ester peaks in the chromat
11.ASSAY
a. Material
Ammonium chloride pH 10 buffer solution
Dissolve 5.4 g of ammonium chloride in water, add 20 mL of ammonium hydroxide and dilute with
water to 100 mL.
Butyl alcohol : Dehydrated alcohol (1 : 1)
Ammonium hydroxide
0.1 M Edetate disodium VS
Eriochrome black TS
0.1 M Zinc sulfate VS

b. Procedure
Accurately weigh 500 mg of sample. Quantitatively transfer into 250-mL Erlenmeyer flask. Add 50
mL of a mixture of butyl alcohol : dehydrated alcohol (1 : 1), 5 mL of ammonium hydroxide, 3 mL of
ammonium chloride pH 10 buffer solution, 30 mL 0.1 M edetate disodium VS, and 1 or 2 drops of
eriochrome black TS, and mix.
Heat at 45o-50oC until the solution is clear. Cool.
Titrate the excess edetate disodium with 0.1 M zinc sulfate VS until the solution color changes from
blue to violet.
Perform a blank determination and make any necessary correction.
Each mL of 0.1 M edetate disodium is equivalent to 2.431 mg of magnesium
Calculation
Concentration of Magnesium =
1
(Vbl - Vspl) x NF x 2.431 x 100 - %LOD x 100% = ...%
(W spl x
100 )
Where:
Vbl = Volume of 0.1 M zinc sulfate VS consumed by 0.1 M edetate disodium VS in the blank
titration (mL)
Vspl = Volume of 0.1 M zinc sulfate VS consumed by 0.1 M edetate disodium VS in the sample
titration (mL)
Wspl = Weight of sample (mg)
%LOD = Percentage of loss on drying obtained from Loss on Drying test (%)
NF = Normality factor of 0.1 M zinc sulfate VS
Prepared by: Checked/Approved by:

Lina Herlina Ponelta Bangun

QC Unit Head QC Responsible Pharmacist