standard testing procedure Bromhexine Hydrochloride

Storage Requirements:
Store protected from light and moisture.
Sampling:
Sample equal quantity from each container / bag. Collect a minimum of 5g from each container/bag sample into individual, self –sealing clear polythene bag kept in another clear self sealing polythene bag bearing ‘Sample for Analysis,lable.After completion of sampling return rest sample on same container. Collect control sample in Pet Bottle/Glass Bottle.
Quantity of Composite Sample:
10 g
Quantity of individual Identification
0.0100 g from each and every container /bag.
Quantity of Control Sample:
2 X 10 g

standard testing procedure Bromhexine Hydrochloride

Description: A white or almost white, crystalline powder; odourless or almost odourless.
Solubility: Sparingly soluble in ethanol (95 per cent); and in methanol; slightly soluble in chloroform;
Practically insoluble in water.
Identification:
Test A may be omitted if tests B, C, D and E are carried out. Tests B, C and D may be omitted if tests A and E are carried out.
A. By IR
B. In the test for related substances, the principal peak in the chromatogram obtained with test solution
Corresponds to that in the chromatogram obtained with reference solution (b).
C. A 2.0 per cent w/v solution of chloramines T and shake; a brownish yellow colour is produced in the
lower layer.
D. A solution prepared by dissolving about 1 mg in 3ml of 0.1 M hydrochloric acid gives the
Reaction for primary aromatic amines (2.3.1)
E. The solution gives reaction A of chlorides
Related Substances: Determine by liquid chromatography
I) Single Maximum Impurity: NMT – 0.2%.
II) Any other Impurity: NMT – 0.1%.
III) Total Impurity: NMT – 0.3%.
Sulphated Ash: NMT 0.1 % w/w
Loss on Drying: NMT 1.0 % w/w
Assay: 98.5% to 101.5%w/w on the dried basis.

analysis of Bromhexine Hydrochloride

Description: A white or almost white, crystalline powder; odourless or almost odourless.
Reporting: Report as Complies/Does not comply.

Solubility: Sparingly soluble in ethanol (95 per cent) and in methanol, Slightly soluble in chloroform;
Practically insoluble in water.
Reporting: Report as Complies/Does not comply.

Identification: Test A may be omitted if tests B, C, D and E are carried out. Tests B, C and D may be omitted if tests A and E are carried out.

A. By IR: Determine by infrared absorption spectrophotometer. Compare the spectrum with that obtained with
Bromhexine hydrochloride RS.
B. In the test of related substances, the principal peak in the chromatogram obtained with test solution corresponds to that
in the chromatogram obtained with reference solution (b).
C. Dissolve about 25mg in a mixture of 1 ml of 1M sulphuric acid and 50ml of water, add 2 ml of dichloromethane and
5ml of a freshly prepared 2.0 per cent w/v solution of chloramineT and shake; a brownish yellow colour is produced in
the lower layer.
D. A solution prepared by dissolving about 1 mg in 3 ml of 0.1 M hydrochloric acid gives the reaction for primary
aromatic amines.
E. Dissolve 20 mg of the sample under examination in 2 ml of water or use 2 ml of the prescribed solution. Acidify with
dilute nitric acid, add 0.5 ml of silver nitrate solution, shake and allow to stand; a curdy white precipitate is formed,
which is insoluble in nitric acid but soluble, after being well washed with water, in dilute ammonia solution, from
which it is reprecipitated by the addition of dilute nitric acid.

Reporting: Report as Complies/Does not comply.

Related Substances: Determine by liquid chromatography
Test solution: Dissolve 50 mg of the substance under examination in 10 ml of methanol.
Reference solution (a): Dissolve 5mg of Bromhexine impurity C RS in methanol, add 1.0 ml of the test solution and dilute to 10.0 ml with the same solvent.
Reference solution (b): A 0.0005 % w/v solution of Bromhexine hydrochloride RS in methanol. (Dissolve 50mg of the Bromhexine hydrochloride RS in 100.0ml of the mobile phase. Take 1.0ml of this solution and dilute to 100ml with mobile phase).
Reference solution (b1): Precede same as reference solution (b).
Chromatographic system
– a stainless steel column 12 cm x 4.6 mm, packed with endcapped octadecylsilane bonded to porous silica (3µm),
– mobile phase: a mixture of 20 volumes of buffer solution prepared by dissolving 0.5 ml of Orthophosphoric acid to 950.0 ml of water, adjust the pH to 7.0 with triethylene, dilute to 1000 ml and 80 volumes of acetonitrile.
– flow rate 1ml per minute
– Spectrophotometer set at 248nm
– Injection volume. 10µl
The relative retention time with reference to Bromhexine hydrochloride for Bromhexine hydrochloride impurity A is about 0.1, for bromhexine hydrochloride impurity B is about 0.2, for bromhexine hydrochloride impurity C is about 0.4 and for bromhexine hydrochloride impurity D is about 0.5.
Inject reference solution (a) the test is not valid unless the resolution between the peak due to bromhexine hydrochloride and bromohexine hydrochloride impurity C is not less than 12.0.
Inject the test solution and reference solution (b). In the chromatogram obtained with test solution, the area of any secondary peak is not more than twice the area of the principal peak in the chromatogram obtained with reference solution (b) (0.2 per cent), the area of one such peak is not more than the area of the principal peak in the chromatogram obtained with reference solution (b) (0.1 per cent) and sum of area of all secondary peaks is not more than 3 times the area of the principal peak in the chromatogram obtained with reference solution (b) (0.3 per cent). Ignore any peak with an area less than 0.5 times the area of the principal peak I the chromatogram obtained with reference solution ( b) (0.05per cent).
Calculation:

1) % of Single Max. Impurity:

Single max Imp area in test solution x std. wt (mg) x 1 x 10 x P x 100

Mean area of reference solution (b) x 100 x 100 x spl. Wt (mg) x 100

2) % of any other Impurity:

Any other Imp area in test solution x std. wt (mg) x 1 x 10 x P x 100

Mean area of reference solution (b) x 100 x 100 x spl. Wt (mg) x 100

3) % of total Impurity:

Total Imp area in test solution x STD wt. (mg) x 1 x 100 x P x 100

Mean area of standard solution (b) x 100 x 100 x spl wt. (mg) x 100

Where, P = Purity of working standard on as such basis
System suitability requirement:
1] The relative standard deviation for the peak area response of Bromhexine hydrochloride for replicate
Injection of reference Solution (b) and replicate injection of reference solution (b) including bracketing standard is not
More than 5.0 % each and relative standard deviation for retention time for the peak of Bromhexine hydrochloride
for replicate Injection of reference solution (b) and replicate injection of reference solution (b) including bracketing
Standard is not more than 2.0 %.
2] The similarity factor of Reference solution (b) and reference solution (b1) is between 0.95 to 1.05,

Calculate similarity factor using formula.

Average peak area response Weight of reference solution (b)

in reference solution (b) in mg

=————————————————————— X ———————————————————-

Peak area response Weight of reference solution (b1)

Obtained with reference solution (b1) in mg

If the similarity factor does not fall within 0.95 to 1.05, prepare fresh reference solution (b) in duplicate,
Re-inject in single injection and calculate similarity factor again as above. If the similarity factor falls
Within the limit, inject the re-prepared reference solution (b) in replicate and continue the sequence for
blank and sample.
3] The column efficiency for the peak of Bromhexine hydrochloride obtained in the chromatogram of individual
reference Solution (b) injected in replicates (including bracketing standard ) and reference solution (b) is not less then
2000 theoretical plates. (Report the value obtained from the fist injection of replicate of reference Solution (b) as well as
last bracketing standard).
Reporting: Report in %.
Sulphated Ash:
Determined 1.0 gm sample in silica crucible ignite for 800ºC and cool add 1 ml of sulphuric acid or ignite & calculated ash.

Calculation:

% of Ash = Weight of residue x 100

Weight of sample

Reporting: Report in % w/w.

Loss on Drying: NMT 1.0 per cent, determined on 1.0 g by drying in an oven at 105°.

Calculation:

% of LOD= Loss on weight x 100

Before drying weight

Reporting: Report in % w/w.

Assay:
Weigh accurately about 0.3 g, dissolve in 70 ml of ethanol (95 per cent), add 1 ml of 0.1 M hydrochloric acid and add titrate with 0.1M sodium hydroxide, determining the end point Potentiometrically. Record the volume added between the two inflections.
1 ml of 0.1 M sodium hydroxide is equivalent to 0.04126 g of C14H20Br2N2, HCl.

Calculation:

Titrate volume x Normality of 0.1M sodium hydroxide x 0.04126 x 100 x 100

0.1x Spl. wt. x (100-LOD)

Reporting: Reporting as per %.

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