Granisetron Injection

Granisetron Hydrochloride Injection

Granisetron Injection is a sterile solution of Granisetron in Water for Injections. It may contain suitable preservatives.

Granisetron Injection contains not less than 93.0 per cent and not more than 107.0 per cent of the stated amount of granisetron, C₁₈H₂₄N₄O.

Usual  strengths. 1 mg per ml; 1 mg per 2 ml; 1 mg per 3 ml; 1 mg per 5 ml; 3 mg per ml; 3 mg per 3 ml.

Identification

2. Determine by thin-layer chromatography (2.4.17), coating the plate withsilica gelGF₂₅₄.

Mobile phase. A mixture of 60 volumes of methylene chloride, 40 volumes of alcohol, 5 volumes of water and 
2 volumes of ammonium hydroxide.

Test solution. Use the undiluted Injection.

Reference solution. Dissolve a quantity of granisetron hydrochloride RS in water to get similar concentration of the test solution.

Apply to the plate 20 µl each of the reference solution and the test solution. Allow the mobile phase to rise 15 cm. Dry the plate under a current of warm air for about 5 minutes and examine the plate under wavelength at 254 nm. The principal spot in the chromatogram obtained with the test solution corresponds to the spot in the chromatogram obtained with the reference solution.

1. In the Assay, the principal peak in the chromatogram obtained with the test solution corresponds to the peak in the chromatogram obtained with reference solution (a). 

Tests

pH (2.4.24). 4.0 to 6.0.

Related substances. Determine by liquid chromatography (2.4.14).

note: Perform the determination under subdued light and use amber autosampler vials and low-actinic glassware.

Buffer solution. Dissolve 15.6 g of monobasic sodium phosphate dihydrate in 900 ml of water, adjust to pH 2.0 with orthophosphoric acid  and dilute with water to 1000 ml.

Test solution. Dilute the sample with water if necessary to obtain a solution having a known concentration of 0.02 per cent w/v of granisetron.

Reference solution (a). A 0.0022 per cent w/v solution of granisetron hydrochloride RS in water.

Reference solution (b). A solution containing 0.0001 per cent w/v of each granisetron impurity B, C and D in a mixture of 75 volumes of water and 25 volumes of methanol.

Chromatographic system

     –   a stainless steel column 15 cm × 4.6 mm, polar end-capped packed with octadecylsilane bonded to porous silica (4 µm),

     –   mobile phase: a mixture of 75 volumes of buffer solution, 24 volumes of methanol and 1.1 volume of tetrahydrofuran,

     –   flow rate: 1.2 ml per minute,

     –   spectrophotometer set at 300 nm,

     –   injection volume: 15 µl.

Name                                              Relative           Correction

                                                    retention time          factor

Granisetron impurity A1            0.5 to 0.6                 ---

Granisetron impurity B2                  0.7                     1.25

Granisetron (Retention time:

about 7.0 minutes)                           1.0                       ---

Granisetron impurity C3                  1.2                      1.0

Granisetron impurity D4            2.1 to 2.3               0.67 

¹ 2-methyl-N-[(1R,3r,5S)-9-methyl-9-azabicyclo[3.3.1]non-3-yl]-2H-indazole-3-carboxamide,

² N-[(1R,3r,5S)-9-methyl-9-azabicyclo[3.3.1]non-3-yl]-1H-indazole-3-carboxamide,

³ N-[(1R,3r,5S)-9-azabicyclo[3.3.1]non-3-yl]-1-methyl-1H-indazole-3-carboxamide,

⁴ 1-methyl-1H-indazole-3-carboxylic acid,

Inject  reference solution (b). The test is not valid unless the resolution between the peaks corresponding to impurity C and granisetron is not less than 2.

Inject reference solution (a) and the test solution. In the chromatogram obtained with the test solution, the area of peak corresponding to impurity C is not more than 0.07 times the area of the principal peak in the chromatogram obtained with reference solution (a) (0.7 per cent). The sum of areas of all the known impurities peaks is not more than 0.13 times the area of the principal peak in the chromatogram obtained with reference solution (a) (1.3 per cent). Other secondary peak is not more than 0.05 times the area of the principal peak in the chromatogram obtained with reference solution (a) 
(0.5 per cent). Ignore any peak with relative retention time of about 0.5 to 0.6 for granisetron impurity A. Ignore any peak with an area less than 0.01 times of the peak obtained from reference solution (a) (0.1 per cent).

Bacterial endotoxins(2.2.3). Not more than 25 Endotoxin unit per mg of granisetron.

Other tests. Comply with the tests stated under Parenteral Preparations (Injections).

Assay. Determine by liquid chromatography (2.4.14), as described under related substances with the following modification.

Test solution. A 0.0022 per cent w/v solution of Granisetron Hydrochloride in water.

Inject reference solution (a) and the test solution.

Calculate the content of C₁₈H₂₄N₄O  in the injection.

Storage. Protected from light. Store at temperature not exceeding 30⁰.

Labelling. The label states (1) the strength in terms of the equivalent amount of granisetron in a suitable dose-volume; (2) the route of injection; (3) that a solution containing visible solid particles must not be used.