Naphazoline Hydrochloride and Pheniramine Maleate Ophthalmic Solution
» Naphazoline Hydrochloride and Pheniramine Maleate Ophthalmic Solution is a sterile, buffered solution of Naphazoline Hydrochloride and Pheniramine Maleate in water adjusted to a suitable tonicity. It contains not less than 90.0 percent and not more than 110.0 percent of the labeled amount of naphazoline hydrochloride (C14H14N2·HCl) and pheniramine maleate (C16H20N2·C4H4O4). It contains a suitable preservative.
Packaging and storage—
Preserve in tight containers, and store at a temperature between 20
and 25, protected from light.
and 25, protected from light.
Identification—
A:
Proceed as directed in the following thin-layer chromatographic procedure.
Naphazoline hydrochloride standard solution—
Dissolve a quantity of USP Naphazoline Hydrochloride RS in water to obtain a solution containing about 1.5 mg per mL.
Pheniramine maleate standard solution—
Dissolve a quantity of USP Pheniramine Maleate RS in water to obtain a solution containing about 6.0 mg per mL.
Test solution—
Dilute, if necessary, a volume of Ophthalmic Solution with water to obtain a solution containing about 0.25 mg of naphazoline hydrochloride per mL and 3 mg of pheniramine maleate per mL.
Procedure—
Separately apply 5 µL of Naphazoline hydrochloride standard solution, 10 µL of Pheniramine maleate standard solution, and 30 µL of the Test solution to a 20-cm × 20-cm thin-layer chromatographic plate (see Chromatography 
621
) coated with a 0.25-mm layer of silica gel. Allow the spots to dry, then place the plate in a saturated chromatographic chamber, and develop in a solvent system consisting of methanol, water, and acetic acid (8:1:1) until the solvent front has moved to about 1.5 cm from the top of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray with ninhydrin TS, and place in an oven at 105 to visualize the spots. Both the naphazoline and pheniramine spots are purplish grey in color. The RF values of the spots obtained from the Test solution correspond to those obtained from the Naphazoline hydrochloride standard solution and the Pheniramine maleate standard solution.
621) coated with a 0.25-mm layer of silica gel. Allow the spots to dry, then place the plate in a saturated chromatographic chamber, and develop in a solvent system consisting of methanol, water, and acetic acid (8:1:1) until the solvent front has moved to about 1.5 cm from the top of the plate. Remove the plate from the developing chamber, mark the solvent front, and allow to air-dry. Spray with ninhydrin TS, and place in an oven at 105 to visualize the spots. Both the naphazoline and pheniramine spots are purplish grey in color. The RF values of the spots obtained from the Test solution correspond to those obtained from the Naphazoline hydrochloride standard solution and the Pheniramine maleate standard solution.
B:
The retention times of the major peaks in the chromatogram of the Assay preparation correspond to those of the Standard preparation, as obtained in the Assay.
Sterility 71—
It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
71—
It meets the requirements when tested as directed for Membrane Filtration under Test for Sterility of the Product to be Examined.
pH 791:
between 5.7 and 6.3.
791:
between 5.7 and 6.3.
Assay—
Buffer solution—
Dissolve 14.2 g of anhydrous dibasic sodium phosphate and 20 mL of triethylamine in 1900 mL of water, adjust with phosphoric acid to a pH of 5.6 ± 0.1, dilute with water to make 2000 mL of solution, and mix.
Mobile phase—
Prepared a filtered and degassed mixture of Buffer and acetonitrile (80:20). Make adjustments if necessary (see System Suitability under Chromatography 621).
621).
Naphazoline hydrochloride stock standard solution—
Dissolve an accurately weighed quantity of USP Naphazoline Hydrochloride RS in Mobile phase to obtain a solution having a known concentration of about 0.75 mg per mL.
Pheniramine maleate stock standard solution—
Dissolve an accurately weighed quantity of USP Pheniramine Maleate RS in Mobile phase to obtain a known concentration of about 3.00 mg per mL.
Standard preparation—
Transfer 1.0 mL of Naphazoline hydrochloride stock standard solution and 3.0 mL of Pheniramine maleate stock standard solution to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix to obtain a solution having known concentrations of naphazoline hydrochloride and pheniramine maleate of 0.03 and 0.36 mg per mL, respectively.
Assay preparation—
Transfer an accurately measured volume of Ophthalmic Solution, equivalent to about 0.75 mg of naphazoline hydrochloride and 9.0 mg of pheniramine maleate, to a 25-mL volumetric flask, dilute with Mobile phase to volume, and mix.
Chromatographic system (see Chromatography 621)—
The liquid chromatograph is equipped with a 270-nm detector and a 4.6-mm × 15-cm column that contains packing L7. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the resolution, R, between the naphazoline peak and the pheniramine peak is not less than 2; the column efficiency, determined from the naphazoline and pheniramine peaks, is not less than 750 theoretical plates; the tailing factor is not greater than 2.5 for pheniramine; and the relative standard deviation for replicate injections is not more than 2.0%.
621)—
The liquid chromatograph is equipped with a 270-nm detector and a 4.6-mm × 15-cm column that contains packing L7. The flow rate is about 1.5 mL per minute. Chromatograph the Standard preparation, and record the peak responses as directed for Procedure: the resolution, R, between the naphazoline peak and the pheniramine peak is not less than 2; the column efficiency, determined from the naphazoline and pheniramine peaks, is not less than 750 theoretical plates; the tailing factor is not greater than 2.5 for pheniramine; and the relative standard deviation for replicate injections is not more than 2.0%.
Procedure—
Separately inject equal volumes (about 25 µL) of the Standard preparation and the Assay preparation into the chromatograph, record the chromatograms, and measure the responses for the peaks. Calculate the quantity, in mg, of naphazoline hydrochloride (C14H14N2·HCl) in each mL of the Ophthalmic Solution taken by the formula:
25(C/V)(rU / rS)
in which C is the concentration in mg per mL of USP Naphazoline Hydrochloride RS in the Standard preparation; V is the volume, in mL, of Ophthalmic solution taken; and rU and rS are the naphazoline peak responses obtained from the Assay preparation and the Standard preparation, respectively. Calculate the quantity, in mg, of pheniramine maleate (C16H20N2·C4H4O4) in each mL of the Ophthalmic Solution taken by the same formula, changing the terms to refer to pheniramine maleate.
Auxiliary Information—
Please check for your question in the FAQs before contacting USP.
Chromatographic Column—
| Topic/Question | Contact | Expert Committee |
| Monograph | Feiwen Mao, M.S.
Scientist 1-301-816-8320 |
(MDOOD05) Monograph Development-Ophthalmics Oncologics and Dermatologicals |
| Reference Standards | Lili Wang, Technical Services Scientist 1-301-816-8129 RSTech@usp.org |
|
| 71 |
Radhakrishna S Tirumalai, Ph.D.
Senior Scientist 1-301-816-8339 |
(MSA05) Microbiology and Sterility Assurance |
USP32–NF27 Page 3034
Pharmacopeial Forum: Volume No. 28(6) Page 1825
Chromatographic columns text is not derived from, and not part of, USP 32 or NF 27.