standard test procedure Ondansetron injection

1.0 OBJECTIVE
1.1 To lay down a procedure for Analysis of Ondansetron injection (1ml)

2.0 SCOPE
2.1 This procedure is applicable to the Analysis of Ondansetron injection (1ml) in quality control,

3.0 RESPONSIBILITY
3.1 Q.C- Chemist

4.0 ACCOUNTABILITY
4.1 Head-Quality Assurance

5.0 Identification: In the assay the principal peak in the chromatogram obtained with the test solution corresponds to

the peak in the chromatogram obtained with the reference solution.
6.0 Description: Pour 10ml finish sample in Beaker and observed Visually.

7.0 pH: Taken 40ml sample in beaker the pH electrode first with purified water followed by sample dip the electrode in sample and observe the pH.

8.0 Extractable volume: Measured the volume by 10 ml Measuring cylinder and determine the volume.

9.0 Particulate matter: Injections that are solutions, when examine under suitable conditions of visibility are clear and

practically free from particles that can be observed on visual inspection by the unaided eye.

10.0Sterility: – Injection comply with the test for sterility as per SOP no. (FPPL/MB/031-00)

11.0 Bacterial endotoxins: –
NMT 9.9 Endotoxins unit/mg of Ondansetron as per SOP no. (FPPL/MB/030-00)

12.0 Related substance: Determine by liquid chromatography.
Test solution: -Taking a measured volume of the injection containing about 2 mg of ondansetron to a 25 ml of volumetric flask dilute to volume with mobile phase.
Reference solution (a) A 0.01 % solution of the Ondansetron hydrochloride RS in the mobile phase
Reference solution (b) dilute 1 ml of reference solution (a) to 100 ml with the mobile phase.
Reference solution (c) A solution containing 0.01% w/v of ondansetron hydrochloride RS and 0.005% w/v

of 3(Dimethylaminomethyl)-1,2,3,9-tetrahydro-9-methyl-4H-Carbozole-4-one RS (Ondansetron hydrochloride A RS) in the mobile phase.

Buffer: – 2.72 gm. potassium dihydrogen phosphate. 1000 ml water. adjust to pH 5.4
With Sodium hydroxide.

Mobile phase: – Buffer: Acetonitrile
50 : 50

Chromatographic condition: – Wavelength-216 nm
Flow rate 1.5 ml /mint.
Injection volume- 10 micro liter.
Column- C18 20cm x 4.6 mm (5 micron) with chemically bonded nitrile group.

Inject reference solution(c) the test is not valid unless the relative retention time and for ondansetron impurity A is

about 1.1. the resolution between ondansetron and ondansetron impurity A is not less than 1.5.

Inject reference solution(a) the tailing factor is not more than 2.0 and the relative standard deviation for replicate injection is not more than 2.0.

13.0 Composition: –
Each ml contains:
Ondansetron hydrochloride I.P.
Eq. to Ondansetron 2.0 mg.

14.0 Assay: – NLT 95.0% and NMT 105.0%
Method of Ondansetron: – Determine by liquid chromatography.
Buffer: 2.72 gm. potassium dihydrogen phosphate. 1000 ml water. adjust to pH 5.4
With Sodium hydroxide.

Mobile phase: – Buffer: Acetonitrile
50 : 50

Standard preparation: -Weight accurately about 50 mg Working standard of Ondansetron in 100 ml of mobile phase Further dilution 5.0 ml to 50 ml with mobile phase.

Sample preparation: -Take sample equivalent 4.0 mg of Ondansetron. Dilute 100 ml of mobile Phase.

Chromatographic condition: –
Wavelength-216 nm
Flow rate 1.5 ml /mint.
Injection volume- 10 micro liter.
Column- C18 25cm x 4.6 mm (5 micron) with chemically bonded nitrile group.

Record the Chromatogram and measure the peak response of the measure peaks. Calculate the content of Ondansetron Respectively.

Retention time of the Principal peak is about 4.3 minutes.

Procedure: – Inject Blank in single injection, Standard preparation in Replicates of three, test preparation in duplicate injection.

System suitability: – % RSD of three replicate injections should not be more than 2.0.

Calculation: –

Area of spl Wt. of std

………………… X……………………. X 100 X % Potency of Standard X 1

Area of std Wt. of spl

15.0 ABBREVIATION: –

Sr. No.	Abbreviation used	Full form of abbreviation used
1.0	STP	Standard Testing Procedure
2.0	QA	Quality assurance
3.0	STD	Standard
4.0	SPL	Sample
5.0	NM	Nano Meter