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  • Title: Validated specific HPLC-DAD method for simultaneous estimation of paracetamol and chlorzoxazone in the presence of five of their degradation products and toxic impurities.
    Author: El-Yazbi AF, Guirguis KM, Bedair MM, Belal TS.
    Journal: Drug Dev Ind Pharm; 2020 Nov; 46(11):1853-1861. PubMed ID: 32894703.
    Abstract:
    This work demonstrates a specific and reliable HPLC with diode array detection (DAD) method for the simultaneous estimation of paracetamol (PAR) and chlorzoxazone (CZ) in the presence of five of their degradation products and toxic impurities; namely; 4-aminophenol (AP), 4-nitrophenol (NP), acetanilide (AT), 4-chloroacetanilide (CA) and 2-amino-4-chlorophenol (ACP). Successful chromatographic separation was accomplished using Waters Symmetry C8 column (3.9 × 150 mm, 5 μm) with gradient elution of the mobile phase consisting of 0.05 M phosphate buffer pH 7.5 and methanol. The gradient elution started with 5% (by volume) methanol ramped up linearly to 50% in 10 min, and then maintained at this percentage afterward till the end of the run. The mobile phase was pumped at a flow rate of 1.0 mL/min. The multiple wavelength detector was adjusted at 244 and 285 nm to quantify PAR and CZ, respectively. Additionally, the wavelength 270 nm was found suitable for monitoring the separation of the entire mixture of PAR, CZ, and their impurities. Seven peaks eluted with excellent resolution at retention times 3.4, 5.7, 8.0, 10.1, 10.8, 13.5, and 14.4 min for AP, PAR, NP, AT, ACP, CZ, and CA, respectively. Performance of the proposed method was validated with respect to linearity, range, precision, accuracy, specificity, robustness, detection, and quantitation limits. Calibration curves were linear in the ranges of 10-75 and 10-100 µg/mL for PAR and CZ, respectively with correlation coefficients not less than 0.9998. The proposed method proved to be specific and stability indicating by the resolution of both drugs from their degradation products and toxic impurities. Validated HPLC method was successfully applied to the analysis of PAR and CZ in their combined capsules dosage form, and assay results were favorably compared with a published reference HPLC method. DAD served as an efficient tool for peak identity and purity verification.
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