standard testing procedure tramadol 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

Description: A white or almost white, crystalline powder.

Solubility: Freely soluble in water and in methanol; very slightly soluble in acetone

Identification:
Test B may be omitted if tests A and C are carried out. Test A may be omitted if tests B and C are carried out.
A By IR
B. In the test for impurity E, the principal spot in the chromatogram obtained with test solution (b)
Corresponds to the principal spot in the chromatogram obtained with reference solution (a).
C. Gives reaction (a) of chlorides.

Appearance of solution: A 5 per cent w/v solution is clear and colourless.

Acidity: To change the colour of the indicator to yellow.

Specific optical rotation: -0.10° to +0.10°.

Impurity E.: The chromatogram obtained with reference solution (c) shows 2 clearly separated spots. In the chromatogram obtained with test solution (a) any secondary spot corresponding to tramadol impurity E is not more intense than the spot in the chromatogram obtained with reference solution (b).

Related Substances: In the chromatogram obtained with the test solution, the area of the peak due to tramadol impurity A is not more than the area of the principal peak in the chromatogram obtained with reference solution (a), the area of any other secondary peak in the chromatogram obtained with reference solution (a). The sum of all the secondary peaks is not more than twice the area of the principal peak in the chromatogram obtained with reference solution (a).Ignore any peak with an area less than 0.1 times the area of the principal peak in the chromatogram obtained with reference solution (a).

Heavy metals: NMT 20 ppm.

Sulphated ash: NMT 0.1%w/w.

Water: NMT 0.5%w/w.

Assay: 99.0% to 101.0%w/w on the anhydrous basis.

Description: A white or almost white, crystalline powder.

Solubility: Freely soluble in water and in methanol
Very slightly soluble in acetone.

Identification:

Test B may be omitted if tests A and C are carried out. Test A may be omitted if tests B and C are carried out.

A By IR
B. In the test for impurity E, the principal spot in the chromatogram obtained with test solution (b)
Corresponds to the principal spot in the chromatogram obtained with reference solution (a).
C. Gives reaction (a) of chlorides.

Appearance of solution: A 5 per cent w/v solution is clear and colourless.

Acidity:To 10ml of 5 per cent w/v solution, add 0.2ml of methyl red solution and 0.2ml of 0.01M hydrochloride hydrochloride acid. The solution is red. Not more than 0.4ml of 0.01M sodium hydroxide is required to change the colour of the indicator to yellow.

Specific optical rotation: Determined on 5.0 per cent w/v solution in water.

Calculation:

SOR = Angle of Rotation X 100
2 X Sample Weight

Impurity E: Determine by thin-layer chromatography

Mobile phase: A mixture of 1 volume of concentrated ammonia, 19 volumes of 2-propanolol and 80 volumes of toluene.

Test solution (a): Dissolve 0.1g of the substance under examination in 2ml of methanol.

Test solution (b): Dilute 1 ml of test solution (a) to 10ml with methanol.

Reference solution (a): A 0.5 per cent w/v solution of tramadol hydrochloride RS in methanol.

Reference solution (b): A 0.1 per cent w/v solution (2 RS)-2-[dimethylamino) methyl] cyclohexane RS (tramadol impurity E RS) in methanol. Dilute 1ml of this solution to 10 ml with methanol.

Reference solution (c): A 0.5 percent w/v solution of (IRS, 2SR)-2-[dimethylamino) methyl]-1-(3- methoxyphenyl) cyclohexanol RS (tramadol impurity A RS) in reference solution (a).

Saturate the plate for 20 minutes with concentrated ammonia. Apply to the plate 10µl of each solution> allow the mobile phase to rise 15cm. Dry the plate in air, expose the plate to iodine vapour for 1hour , and examine in ultraviolet at 254nm. The chromatogram obtained with reference solution (c) shows 2 clearly separated spots. In the chromatogram obtained with test solution (a) any secondary spot corresponding to tramadol impurity E is not more intense than the spot in the chromatogram obtained with reference solution (b) (0.2 per cent).

Related Substances: Determine by liquid chromatography.

Test solution: Dissolve 0.15 g of the substance under examination in 100.0 of the mobile phase.

Reference solution (a): Dilute 2.0 ml of the test solution to 10.0 ml with the mobile phase. Dilute 1.0 ml of this 100.0ml with the mobile phase.

Reference solution (b): Dissolve 5mg of tramadol impurity A RS in 4.0 ml of the test solution and dilute to 100.0 ml with the mobile phase.

Chromatographic system
– a stainless steel column 25 cm X 4.0 mm packed with endcapped octylsilane bonded to porous silica (5µm),
– mobile phase: a mixture of 29.5 volumes of acetonitrile and 70.5 volumes of a mixture of 0.2 ml of trifluoroacetic acid and 100ml of water,
– flow rate 1 ml per minute
– spectrophotometer set at 270nm
– Injection volume. 20µl.

Inject reference solution (b) the test is not valid unless the resolution between the peaks due to tramadol impurity A and tramadol is not less than 2.0. The relative retention time with reference to tramadol for tramadol impurity A is about 0.85

Inject the test solution and reference solution (a). Run the chromatogram 45 times the retention time of the principal peak. In the chromatogram obtained with the test solution, the area of the peak due to tramadol impurity A is not more than the area of the principal peak in the chromatogram obtained with reference solution (a) (0.2 per cent), the area of any other secondary peak in the chromatogram obtained with reference solution (a) (0.1 per cent). The sum of all the secondary peaks is not more than twice the area of the principal peak in the chromatogram obtained with reference solution (a) (0.4 per dent0 Ignore any peak with an area less than 0.1 times the area of the principal peak in the chromatogram obtained with reference solution (a) (0.02 per cent).

Heavy metals: 1.0 g complies with the limit test for heavy metals. Method D (20 ppm).

Sulphated ash: Weight accurately 1.0gm of sample in silica crucible ignite in muffle farness at 700ºC for 120
minutes.

Reporting: Report in percentage.

Assay: Dissolve 0.18g in 25 ml of anhydrous acetic acid and add 10ml of acetic anhydride. Titrate with 0.1M perchloric acid, determining the end-point potentiometrically. Carry out a blank titration.

1 ml of 0.1m perchloric acid is equivalent to 0.02998g of C16H26ClNO2.