The specification and Analytical Testing Procedure (ATP) / Standard Testing Procedure (STP) for the drug product Granisetron Hydrochloride Injection USP (as per USP monograph). The below procedures are most updated and revised.

Specification – Granisetron Hydrochloride Injection USP

Sr. No.	Test Parameter	Specification
1.	Description	Clear colorless solution filled in transparent glass vial with transparent flip off seal (Refer the manufacturer specification)
2.	Identification :
	A)  Granisetron Hydrochloride	By HPLC; The UV Spectrum of major peak of the sample solution corresponds to that of the standard solution, as obtained in the Assay.
	B)  Benzyl Alcohol	By HPLC; The retention time of major peak of the sample solution correspond to that of the standard solution, as obtained in the Assay.
3.	pH	Between 4.00   and   6.00
4.	Average Volume	Not less than 3.00 ml.
5.	Uniformity of Volume	3.00   to   3.45 ml.
6.	Bacterial Endotoxins	Not more than 25 USP Endotoxin Units per mg of Granisetron.
7.	Sterility	Sample should be sterile.
8.	Particulate Matter :
	a)  ≥ 10 µm	Not more then 6,000 particles / container.
	b)  ≥ 25 µm	Not more then 600 particles / container
9.	Related Compounds – By HPLC
	Granisetron Related  Compound C	Not more than 0.7%
	Total Specified Impurities	Not more than 1.3%
	Any Unspecified Impurity	Not more than 0.5%
10.	Assay
	Each ml contains : Granisetron Hydrochloride  USP eq. to  Granisetron  1 mg	0.93   to   1.07 mg 93.00   to   107.00% of labeled amount

Download the Specification – Specification -Granisetron Hydrochloride Injection USP

Standard Testing Procedure – Granisetron Hydrochloride Injection USP

1.0     Description

• • Carry out the test as per procedure. For detailed Description Test Procedure – Refer GTP – Description / Appearance Test.

2.0     Identification

• • Granisetron Hydrochloride – By UV 300nm.

• • Procedure: By HPLC; The UV Spectrum of major peak of the sample solution corresponds to that of the standard solution, as obtained in the Assay. UV Detector range of 200 to 400nm.

• • By HPLC.

• • 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.

• • Benzyl Alcohol – By HPLC; The retention time of principal peak of sample is concordant with the principal peak of standard solution, in Assay procedure.

3.0     pH

• • Carry out the test as per procedure. Refer detailed procedure for pH value determination.

4.0     Average Volume

• • Carry out the test as per procedure.

5.0     Uniformity of Volume  

• • Carry out the test on 5 containers as per procedure.

6.0     Related Compounds – Granisetron Hydrochloric Acid:

• • Note : Perform the determination under subdued light and use amber auto sampler 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 for Related Substances Test – Granisetron Hydrochloric Acid :

  • 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) (C_S/C_T) (r_i /r_s)

• • 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  C_S  is the concentration, in mg per ml, of Granisetron hydrochloride in the Standard solution;  C_T  is the concentration, in mg per ml, of Granisetron in the Test solution, based on the label claim;  r_i  is the peak response of each impurity obtained from the Test solution; and  r_S   is the peak response of the Granisetron peak, obtained from Standard solution.

• • Disregard the peak due to Granisetron related compound A that elutes at the relative retention time of about 0.5 to 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 Factory (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

7.0     Assay of  Granisetron Hydrochloride

• • Procedure  :  By HPLC.

• • Note : Perform the determination under subdued light and use amber auto sampler vials and low-acting glassware.

• • Chromatographic System  : 

• • The liquid chromatograph is equipped with a    300 nm detector and a 4.6 mm × 15 cm column that contain 4 µm polar end-capped 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%.

• • Buffer Solution pH 2.0 :

• • Dissolve 15.6 gm 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.

• • System Suitability Preparation  :  

• • Weigh Granisetron related B, C, & D each 2.0 mg and USP Granisetron Hydrochloride RS 2.2 mg in 20 ml volumetric flask and dissolve in a mixture of water and Methanol (75:25).

• • Take 1 ml solution to 100 ml and make up with water up to the Mark.

• • Standard Preparation :

• • Dissolve an accurately 110.0 mg weighed quantity of USP Granisetron Hydrochloride RS in 100 ml volumetric flask add sufficient water and sonicate to complete dissolve than make up with water up to the mark than filter and use 10 ml of filtrate than makeup 100 ml with water.

• • Assay Preparation  :  Diluted 1ml of injection to  10 ml with water.

• • 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.

• • Calculation :

• • Calculate the percentage of the labeled amount of Granisetron (C18H24N4O) in each ml of injection by the formula :

Spl. Area  x  Std.  Wt. x  Dilu. Factor  x   Potency  x  0.8954

Std. Area  x  Spl. Vol.  x 100

 

=    mg   found

 

% found   =   mg found  x  100

                Claim

• Benzyl Alcohol determination procedure

  • Chromatographic Conditions  :

Mobile phase       :         Buffer  :  Acetonitrile  (70  :  30)

Column                 :         C₁₈,  250 x  4.6 mm, 5m  (Inertsil ODS or equivalent).

Detector                :         UV – At  220 nm.

Flow rate               :         1.0 ml / minute.

Temperature        :         40° C.

Inject volume       :         20 ml.

• • Buffer Preparation :

• • Prepare 0.1 M Ammonium acetate in 1000 ml distilled water, adjust pH 6.5 with Orthophosphoric acid.

• • Standard Preparation  :

• • Pipette out accurately 0.5 ml of Benzyl Alcohol WS, in a clean    and dry 50 ml amber coloured volumetric flask.  Make up the volume with mobile phase and sonicate to dissolve.

• • Cool the flask, if necessary.

• • Further Pipette out 2 ml of the above solution in a 100 ml amber coloured volumetric flask, and make up the  volume with mobile phase to obtain the concentration about 200 mcg/ml.

• • Sample Preparation  : 

• • Transfer completely 10 ml of sample solution in clean and dry 100 ml volumetric flask,  add about 80 ml of mobile phase, shake and make up the volume with mobile phase.

• • Further 5 ml to 25 ml volumetric flask and make up the volume with mobile phase to obtain the concentration about 200 mcg/ml.

• • Separately inject 20 ml of freshly prepared, standard and sample preparation.

• • Record the chromatogram and calculate the assay by given formula.

• • Calculation  :

 

Spl. Area  x   0.5  x    2    x  Potency (aob)  x   100   x   25 .

Std. Area   x  50  x   100     x 10  x  5

=    mg   found

% found   =   mg found x 100

              L. Claim

Download the STP – MOA – Granisetron Hydrochloride Injection USP