HPLC METHOD DEVELOPMENT AND VALIDATION: SIMULTANEOUS DETERMINATION OF ACTIVE INGREDIENTS IN COUGH AND COLD PHARMACEUTICALS
Keywords:
High performance liquid chromatography, Active Ingredients Cough and Cold Pharmaceuticals, ValidationAbstract
Objective: This study aimed to develope a simple reversed-phase high performance chromatographic method for simultaneous determination of pseudoephdrine HCI, pheniramine maleate, acetaminophen, guaifenisin, pyrilamine maleate, chlorpheniramine maleate, triprolidine HCI, dextromethorphan HBr, diphenhydramine HCI in cough and cold pharmaceuticals.
Methods: The separation of these compounds was achieved within 37.9 min on a Nucleodur gravity C18 column (250 x 4.0 mm, 5μm). The chromatographic separation of these compounds performed in a single run by using isocratic mobile phase consisting of methanol:buffer mixture (38:62, v/v) at room temperature, with flow rate of 0.75 mL.min-1.
An ultraviolet absorption at 210 nm was monitored. 2,4,6-trimethoxybenzaldehyde was used as an internal standard (ISTD). The selectivity, linearity of calibration, accuracy, intraday and interday precision and forced degradation studies were examined as parts of the method validation.
Results: The concentration–response relationship was linear over a concentration range of 0.2-250 µg.mL-1 for acetaminophen, 0.5–250 µg.mL-1 for pseudoephdrine HCI and pheniramine maleate, 1–250 µg.mL-1 for guaifenisin, 2.5-250 µg.mL-1 for chlorpheniramine maleate and triprolidine HCI, 5-250 µg.mL-1 for pyrilamine maleate and diphenhydramine HCI, 10-20 µg.mL-1 for dextromethorphan HBr with correlation coefficients better than 0.9993. The relative standard deviations of the intraday and interday were all less than 4%. Conclusion: The proposed liquid chromatographic method was successfully applied for the routine analysis of these compounds in different cough and cold pharmaceutical preparations such as syrups and tablets.
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