Abstract
A reversed phase high performance liquid chromatography (RP-HPLC) method for the simultaneous quantification of bedaquiline (TMC207), moxifloxacin and pyrazinamide in a pharmaceutical powder formulation for inhalation has been developed and validated. The powder was simply dissolved in methanol and the analytes separated in a run time of 20min on a Luna C18 (2) (150x4.6mm, 5 mu m) column using gradient elution with methanol and triethylamine phosphate buffer (pH 2.5) delivered at 1.2mL/min. The detection (with retention time) was carried out at 269nm (2.9min) for pyrazinamide, 296nm (7.0min) for moxifloxacin and 225nm (16.3min) for bedaquiline, respectively. The method was linear for all analytes in the concentration range 1-100 mu g/mL with correlation coefficients >0.998. Lower limits of quantitation (mu g/mL) of bedaquiline, moxifloxacin and pyrazinamide were 0.56, 0.43 and 0.24, respectively. The method was accurate (relative error in the range -0.2 to 2.2) and precise (%RSD 6.2) with recovery in the range 100.0-104.7%. The method was successfully applied to determine the drug content and content uniformity of the three analytes in a spray-dried combination powder formulation for inhalation containing L-leucine.
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