Determination of bromhexine by LC-MS/MS and comparative study of distribution kinetics in rats after inhalation administration
Objective To establish an LC-MS/MS method for the determination of bromhexine in rat plasma and lung,and the distribution in plasma and lung tissue was compared between the test and the reference formulations of bromhexine hydrochloride inhalation solution in rats.Methods Samples were pretreated by a simple protein precipitation method with diphenhydramine as the internal standard.Chromatographic separation was performed on a ZORBAX Eclipse XDB-C18 column using a gradient elution program with solvents consisting of methanol and 0.5％formic acid-5％methanol aqueous solution.The detection was operated in the positive ion mode using multiple reaction monitoring(MRM)with an electrospray ionization source(ESI).The analytical methods for plasma and lung tissue were fully and partially validated,respectively.Healthy Sprague-Dawley rats were divided into two groups randomly and administered by inhalation with test and reference formulations of bromhexine hydrochloride inhalation solution,respectively.The plasma,lung and tracheobronchus were collected at predetermined time points and the concentration of bromhexine was measured.Results Bioanalytical method validation results all met the criteria of the guidelines.The distribution rate and extent of bromhexine in plasma,lung and tracheobronchus between the two formulations were consistent.The relative bioavailability in the main target organ tracheobronchus was about 97.4％.Conclusion The LC-MS/MS method established in this study is rapid,sensitive and simple.The distribution of bromhexine between the two formulations in rat plasma and lung tissue is consistent,which can provide data support for the localization of the new formulation.
LC-MS/MSbromhexineinhalation administrationcomparative distribution kinetics study
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