Granisetron Hydrochloride Injection
» Granisetron Hydrochloride Injection is a sterile solution of Granisetron Hydrochloride in Water for Injection. It contains the equivalent of not less than 93.0 percent and not more than 107.0 percent of the labeled amount of granisetron (C18H24N4O). It may contain suitable preservatives.
Packaging and storage— Preserve in single-dose or multiple-dose containers, protected from light, and store at controlled room temperature.
Labeling— It meets the requirements for Labeling under Injections 1. Label it to indicate the name and the quantity of any added preservative.
USP Reference standards 11
USP Endotoxin RS
USP Granisetron Hydrochloride RS
USP Granisetron Related Compound B RS
(N-[(1R,3r,5)-9-Methyl-9-azabicyclo[3.3.1]non-3-yl]-1H-indazole-3-carboxamide).
USP Granisetron Related Compound C RS
N-[(1R,3r,5S)-9-Azabicyclo[3.3.1]non-3-yl]-1-methyl-1H-indazole-3-carboxamide.
USP Granisetron Related Compound D RS
1-Methyl-1H-indazole-3-carboxylic acid.
Identification
A: Thin-Layer Chromatographic Identification Test 201
Developing solvent— Prepare a mixture of methylene chloride, alcohol, water, and ammonium hydroxide (60:40:5:2).
Standard solution— Dissolve an accurately weighed quantity of USP Granisetron Hydrochloride RS in water or alcohol to obtain a solution having a concentration of granisetron that matches the concentration of granisetron in the Test solution. To calculate the concentration of granisetron in the Standard solution, use the molecular weights of granisetron (312.41) and granisetron hydrochloride (348.87).
Test solution— Use the undiluted Injection.
Procedure— Separately apply equal volumes of the Standard solution and the Test solution, equivalent to about 4 to 5 µg of granisetron, to a suitable thin-layer chromatographic plate coated with a 0.25-mm layer of chromatographic silica gel mixture, dry the spots under a current of warm air for about 5 minutes, and develop the plate in a paper-lined chromatographic chamber equilibrated with Developing solvent prior to use. Allow the chromatogram to develop until the solvent front has moved about 15 cm. Remove the plate, dry the plate under a current of warm air, and examine the plate under short-wavelength UV light: the principal spot from the Test solution corresponds in appearance and RF value to that of the Standard solution.
B: The retention time of the major peak in the chromatogram of the Assay preparation corresponds to that in the chromatogram of the Standard preparation, as obtained in the Assay.
Bacterial endotoxins 85 It contains not more than 25 USP Endotoxin Units per mg of granisetron.
Sterility 71: meets the requirements.
pH 791: between 4.0 and 6.0.
Particulate matter 788: meets the requirements for small-volume injections.
Related compounds— [note—Perform the determination under subdued light, and use amber autosampler vials and low-actinic glassware. ]
Buffer solution pH 2.0, Mobile phase, System suitability preparation, and Chromatographic system— Prepare as directed in the Assay.
Standard solution— Use the Standard preparation prepared as directed in the Assay.
Test solution— Use the undiluted Injection.
Procedure— Separately inject equal volumes (about 15/L µL), where L is the labeled amount, in mg, of granisetron per mL of Injection of the Standard solution and the Test solution, into the chromatograph. Record the chromatograms for at least three times the retention time of the granisetron peak, identify the impurities listed in Table 1, and measure the peak responses. Calculate the percentage of each impurity relative to the labeled content of granisetron in the portion of Injection taken by the formula:
100(1 / F)(312.41 / 348.87)(CS / CT)(ri / rS)
in which F is the relative response factor as listed in Table 1; 312.41 and 348.87 are the molecular weights of granisetron and granisetron hydrochloride, respectively; CS is the concentration, in mg per mL, of granisetron hydrochloride in the Standard solution; CT is the concentration, in mg per mL, of granisetron in the Test solution, based on the label claim; ri is the peak response of each impurity obtained from the Test solution; and rS is the peak response of the granisetron peak, obtained from the Standard solution. Disregard the peak due to granisetron related compound A that elutes at the relative retention time of about 0.5–0.6 as this impurity is controlled in the drug substance monograph. Not more than 0.7% of granisetron related compound C is found, not more than 1.3% of total specified impurities is found, and not more than 0.5% of any unspecified impurity is found. The reporting level for impurities is 0.1%.
Table 1
Name Relative
Retention
Time
Relative
Response
Factor (F)
Granisetron related compound A1 0.5–0.6
Granisetron related compound B2 0.7 0.8
Granisetron 1.0
Granisetron related compound C3 1.2 1.0
Granisetron related compound D4 2.1–2.3 1.5
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.
Other requirements— It meets the requirements under Injections 1.
Assay— [note—Perform the determination under subdued light, and use amber autosampler vials and low-actinic glassware. ]
Buffer solution pH 2.0— Dissolve 15.6 g of monobasic sodium phosphate dihydrate in 900 mL of water, adjust with phosphoric acid to a pH of 2.0, and dilute with water to 1000 mL.
Mobile phase— Prepare a mixture of Buffer solution pH 2.0, methanol, and tetrahydrofuran (75:24:1.1), mix, and degas. Make adjustments if necessary (see System Suitability under Chromatography 621).
System suitability preparation— Dissolve suitable quantities of USP Granisetron Hydrochloride RS, USP Granisetron Related Compound B RS, USP Granisetron Related Compound C RS, and USP Granisetron Related Compound D RS in a mixture of water and methanol (75:25), to obtain a solution containing about 0.1 mg of each component per mL. Dilute with water to obtain a solution containing about L µg of each component per mL, where L is the labeled amount, in mg, of granisetron per mL of Injection.
Standard preparation— Dissolve an accurately weighed quantity of USP Granisetron Hydrochloride RS in water to obtain a solution having a known concentration of about (0.11 × L) mg of granisetron hydrochloride per mL, where L is the labeled amount, in mg, of granisetron per mL of Injection.
Assay preparation— Use the Injection diluted 1:10 with water.
Chromatographic system (see Chromatography 621) The liquid chromatograph is equipped with a 300-nm detector and a 4.6-mm × 15-cm column that contains 4-µm polar endcapped packing L1. The flow rate is about 1.2 mL per minute. Chromatograph the System suitability preparation, and identify the components based on the information listed in Table 1. Record the peak responses as directed for Procedure: the resolution, R, between the granisetron and granisetron related compound C peaks is not less than 2. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the tailing for the granisetron peak is not more than 3; and the relative standard deviation for a minimum of six replicate injections is not more than 2.0%.
Procedure— Separately inject equal volumes (about 15/L µL), where L is the labeled amount, in mg, of granisetron per mL of Injection, of the Standard preparation and the Assay preparation into the chromatograph. Record the chromatograms for at least three times the retention time of the granisetron peak, and measure the responses for the major peaks. Calculate the percentage of the labeled amount of granisetron (C18H24N4O) in each mL of the Injection by the formula:
100(312.41 / 348.87)(C/L)(rU / rS)
in which 312.41 and 348.87 are the molecular weights of granisetron and granisetron hydrochloride, respectively; C is the concentration, in mg per mL, of granisetron hydrochloride in the Standard preparation; L is as defined above; and rU and rS are the peak areas obtained from the Assay preparation and the Standard preparation, respectively.
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