Olopatadine Hydrochloride Ophthalmic Solution

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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1 DEFINITION 

Olopatadine Hydrochloride Ophthalmic Solution is a sterile aqueous solution of Olopatadine Hydrochloride. It contains NLT 90.0% and NMT 110.0% of the labeled amount of olopatadine (C₂₁H₂₃NO₃). It may contain suitable antimicrobial agents. 

2 IDENTIFICATION 

A. The retention time of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. 

B. The UV spectrum in the range of 270–370 nm of the major peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay. 

3 ASSAY 

Procedure 

Protect all solutions containing olopatadine hydrochloride from light. 

Buffer: Dissolve 13.6 g of monobasic potassium phosphate in 1 L of water, add 1 mL of triethylamine, and mix. Adjust with phosphoric acid to a pH of 3.0. 

Mobile phase: Acetonitrile and Buffer (28:72) 

Standard solution: 0.1 mg/mL of USP Olopatadine Hydrochloride RS in Mobile phase 

Sample solution: Nominally equivalent to 0.1 mg/mL of olopatadine in Mobile phase, from Ophthalmic Solution 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 299 nm or diode array. [Note—Use a diode array detector to perform the Identification B test.] 

Column: 4.6-mm × 15-cm; 5-µm packing L7 

Flow rate: 1 mL/min 

Injection volume: 30 µL 

System suitability 

Sample: Standard solution 

Suitability requirements 

Tailing factor: NMT 2.0 

Relative standard deviation: NMT 2.0% 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of the labeled amount of olopatadine (C₂₁H₂₃NO₃) in the portion of Ophthalmic Solution taken: 

Result = (r_U/r_S) × C_S × (1/F) ×  (M_r1/M_r2) 

r_U = peak response from the Sample solution 

r_S = peak response from the Standard solution 

C_S = concentration of USP Olopatadine Hydrochloride RS in in the Standard solution (mg/mL) 

C_U = nominal concentration of olopatadine in the Sample solution (mg/mL) 

M_r1 = molecular weight of olopatadine 337.41

M_r2 = molecular weight of olopatadine hydrochloride,, 373.87

Acceptance criteria: 90.0%–110.0% 

4 IMPURITIES 

Limit of Early Eluting Impurities 

Protect all solutions containing olopatadine hydrochloride from light. 

Mobile phase: Prepare as directed in the Assay. 

System suitability solution: 0.2 mg/mL of USP Olopatadine Hydrochloride RS and 0.02 mg/mL of USP Olopatadine Related Compound B RS in Mobile phase 

Standard solution: 0.2 mg/mL of USP Olopatadine Hydrochloride RS in Mobile phase 

Sample solution: Equivalent to 0.2 mg/mL of olopatadine in Mobile phase, from Ophthalmic Solution 

Blank solution: Mobile phase 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 299 nm 

Column: 4.6-mm × 15-cm; 5-µm packing L7 

Flow rate: 1 mL/min 

Injection volume: 30 µL 

Run time: At least 1.6 times the retention time of the major peak 

System suitability 

Samples: System suitability solution and Standard solution 

Suitability requirements 

Resolution: NLT 2.0 between olopatadine and olopatadine related compound B, System suitability solution 

Relative standard deviation: NMT 2.0%, Standard solution 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of each impurity in the portion of Ophthalmic Solution taken: 

Result = (r_U/r_S) × (C_S/C_U) × (M_r1/M_r2) x (1/F) x 100

r_U = peak response of each impurity from the Sample solution  

r_S = peak response of olopatadine from the Standard solution 

C_S = concentration of USP Olopatadine Hydrochloride RS in in the Standard solution (mg/mL) 

C_U = nominal concentration of olopatadine in the Sample solution (mg/mL) 

M_r1 = molecular weight of olopatadine hydrochloride, 337.41

M_r2 = molecular weight of olopatadine, 373.87

F = relative response factor for each individual impurity (see Table 1) 

Acceptance criteria: See Table 1. Disregard any peaks corresponding to those of the Blank solution and any peaks with a relative retention time, measured with respect to olopatadine, greater than 1.5. Disregard any peak less than 0.1%. 

Table 1 

^a11-[(E)-3-(Dimethylamino)propylidene]-6,11-dihydrodibenzo[b,e]oxepin-2-acetic acid.

^b(Z)-11-[3-(Dimethylamino)propylidene]-6,11-dihydrodibenzo[b,e]oxepine-2-carbaldehyde. 

Limit of Late Eluting Impurities 

Protect all solutions containing olopatadine hydrochloride from light. 

Buffer: Prepare as directed in the Assay. 

Mobile phase: Acetonitrile and Buffer (1:1) 

System suitability solution: 0.02 mg/mL of USP Olopatadine Hydrochloride RS and 0.01 mg/mL of USP Olopatadine Related Compound C RS in Mobile phase 

Standard solution: 0.01 mg/mL of USP Olopatadine Related Compound C RS in Mobile phase 

Sample solution: Use the Sample solution from the test for Limit of Early Eluting Impurities. 

Blank solution: Mobile phase 

Chromatographic system 

(See Chromatography 〈621〉, System Suitability.) 

Mode: LC 

Detector: UV 299 nm 

Column: 4.6-mm × 15-cm; 5-µm packing L7 

Flow rate: 1 mL/min 

Injection volume: 30 µL 

Run time: At least 3 times the retention time of the olopatadine related compound C peak 

System suitability 

Samples: System suitability solution and Standard solution 

[Note—The relative retention times for olopatadine and olopatadine related compound C are 0.3 and 1.0, respectively.] Suitability requirements 

Resolution: NLT 7.0 between olopatadine and olopatadine related compound C, System suitability solution Relative standard deviation: NMT 2.0%, Standard solution 

Analysis 

Samples: Standard solution and Sample solution 

Calculate the percentage of each impurity in the portion of Ophthalmic Solution taken: 

Result = (r_U/r_S) × (C_S/C_U) × 100 

r_U = peak response of each impurity from the Sample solution  

r_S = peak response of olopatadine related compound C from the Standard solution 

C_S = concentration of USP olopatadine related compound C RS in the Standard solution (mg/mL) 

C_U = nominal concentration of olopatadine in the Sample solution (mg/mL) 

Acceptance criteria: See Table 2. Disregard any peaks corresponding to those of the Blank solution and any peaks with a relative retention time, measured with respect to olopatadine related compound C, less than 0.7. Disregard any peak less than 0.1%. 

Table 2 

^aTotal impurities are the sum of olopatadine related compound B, olopatadine related compound C, olopatadine E-isomer, olopatadinecarbaldehyde, and all unspecified impurities found in the tests for Limit of Early Eluting Impurities and Limit of Late Eluting Impurities. 

5 SPECIFIC TESTS 

Sterility Tests 〈71〉: Meets the requirements 

pH 〈791〉: 5.0–8.0 

Osmolality and Osmolarity 〈785〉: 260–340 mOsmol/kg 

6 ADDITIONAL REQUIREMENTS 

Packaging and Storage: Preserve in tight, light-resistant containers. Store between 4° and 25°. 

USP Reference Standards 〈11〉 

USP Olopatadine Hydrochloride RS 

USP Olopatadine Related Compound B RS 

(Z)-3-{2-(Carboxymethyl)dibenzo[b,e]oxepin-11(6H)-ylidene}-N,N-dimethylpropan-1-amine oxide. 

C₂₁H₂₃NO₄ 353.41 

USP Olopatadine Related Compound C RS 

11-Oxo-6,11-dihydrodibenzo[b,e]oxepin-2-yl acetic acid.

 C₁₆H₁₂O₄ 268.26