Abstract
Introduction: A simple rapid and precise HPLC method was developed for estimation of TH in nasal simulated fluid and stability was assessed in various stressed conditions.
 Methods: Chromatographic separation of TH in nasal simulated fluid was done using HPLC AS-4050 coupled with Jasco UV 2075 Plus detector, Jasco LC-Net 11/ADC valve, Jasco PU-2080 pump and hypersil gold C18 (250x6x5 µm) column, ChromNAV 2.0 Chromatography Data System software with mobile phase as acetonitrile: water (65:35) and acetonitrile: NSF (60:40) at a flow rate of 1ml/min and having run time of 10 min with loop volume of 20 µl and detection wavelength of 252 nm. The method was validated for accuracy, precision, linearity, specificity, and sensitivity in accordance with ICH (Q2B) guidelines.
 Results: The results of all the validation parameters were found to be within the acceptable limits. The calibration plots were linear over the concentration ranges from 2 to 14µg/ml. The accuracy and precision were found to be between 97.04±0.112 to101.081±0.0191and ≤2% for three drugs. Developed method was successfully applied for the determination TH in nasal simulated fluid and recovery was found to be >98% for three drugs. The degradation products produced as a result of stress studies did not interfere with drug peak.
 Conclusion: The developed method was found to be simple, specific, economic, reliable, accurate, precise, and reproducible used as a quality control tool for analysis of pure thymoquinone in nasal simulated fluid.
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