Standard testing procedure of Iron Sucrose Injection

1.0 OBJECTIVE
1.1 To lay down a procedure for Analysis of Iron Sucrose Injection

2.0 SCOPE
2.1 This procedure is applicable to the Analysis of Iron Sucrose Injection . in QC lab

3.0 RESPONSIBILITY
3.1 Q.C- Chemist

4.0 ACCOUNTABILITY
4.1 Head-Quality Assurance

5.0 Identification:
(A) Iron: – To 2.5 ml of injection add 17.5 ml of water and 5 ml of hydrochloric acid mix and heat for 5 minutes in a boiling water bath.

Cool and dropwise 13.5 N ammonium hydroxide until no further precipitation of ferric hydroxide occurs and filter wash

the precipitate with water to remove excess ammonium hydroxide, dissolve the precipitate in Aluminium volume

of 2 N hydrochloric acid, and add sufficient water to make a volume of 20 ml .to 3 ml of the solution so obtained

add 1 ml of potassium thiocyanate the resulting solution is red. To 1 ml of solution add 5 ml of amyl alcohol or

ethyl ether shake and allow to stand the organic layer is pink. To a separate 1 ml aliquot of solution add 2 ml of mercuric chloride the red colour is discharged iron (iii).

(B) Sucrose: – 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

11.0 Bacterial endotoxins: – Not more than 3.7 USP Endotoxin per mg. of iron sucrose as per SOP

12.0 Composition: –
Each 5 ml contains:
Ferric hydroxide in complex with sucrose
Eq. to elemental iron 100 mg.

13.0 Assay: –

Method of Iron: – Determine by atomic absorption spectrophotometry.
Transfer about 350mg of ferrous ammonium sulfate, accurately weighed, to a 100ml volumetric flask add water to dissolve, dilute with water to volume and mix to obtain a solution having a concentration of about 50µg of iron per ml.
Transfer2.64g of calcium chloride to a 100ml volumetric flask add 500ml of water, and swirl to dissolve add 5.0ml to hydrochloride acid and dilute with water to volume.
To separate 50ml volumetric flasks transfer 2.0 ,4.0 ,6.0 ,8.0, and 10.0ml of iron stock solution dilute each flask with calcium chloride solution to volume and mix to obtain standard preparations having known concentration of about 2.0 ,4.0 ,6.0 ,8.0 and 10.0µg of iron per ml.

Sample preparation: -Take sample equivalent about 2.0 ml injection of 100 ml volumetric flask add 5 ml of hydrochloric acid until the solution turns yellow dilute with the calcium chloride further dilution 2.0 ml to 100 ml volumetric flask with calcium chloride.

Chromatographic condition: -Wavelength-248.3 nm with the suitable atomic absorption spectrophotometry.

14. Assay: –
Method of sucrose: – Determine by liquid chromatography.
Mobile phase: Acetonitrile: water.
79 : 21

Standard preparations— Dissolve an accurately weighed quantity of USP Sucrose RS in water, and quantitatively dilute with water to obtain solutions having known concentrations of about 13, 16, 18, 21, and 23 mg of sucrose per ml.

Sample preparation: -Transfer about 1.875 g of Injection, accurately weighed, into a 25-mL flask, add 1.25 mL of water, and mix. Add 1.25 mL of a monobasic sodium phosphate solution, prepared by dissolving 30 g in 50 mL, and mix. Allow the resulting solution to stand for 10 minutes to precipitate the colloidal ferric hydroxide. Dilute with water to volume, and mix. Centrifuge this solution at 3000 rpm for 15 minutes. Pass the resulting solution through a filter, discarding the first 2 mL of the filtrate.

Chromatographic condition: -The liquid chromatograph is equipped with a column compartment and a refractive index detector each maintained at a controlled temperature between 20 and 25 (±2) and a 4-mm × 25-cm column that contains packing L8. The flow rate is about 2.0 mL per minute. Chromatograph the Standard preparations, and measure the peak areas as directed for Procedure. The correlation coefficient obtained from the linear regression of the Standard preparations is not less than 0.998. [NOTE—The retention time for sucrose is about 8 minutes.

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