Development and validation of a RP-HPLC method for simultaneous quantification of bedaquiline (TMC207), moxifloxacin and pyrazinamide in a pharmaceutical powder formulation for inhalation

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 20 min on a Luna C18 (2) (150 × 4.6 mm, 5 µm) column using gradient elution with methanol and triethylamine phosphate buffer (pH 2.5) delivered at 1.2 mL/min. The detection (with retention time) was carried out at 269 nm (2.9 min) for pyrazinamide, 296 nm (7.0 min) for moxifloxacin and 225 nm (16.3 min) for bedaquiline, respectively. The method was linear for all analytes in the concentration range 1-100 µg/mL with correlation coefficients >0.998. Lower limits of quantitation (µg/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.

GRAPHICAL ABSTRACT

KEYWORDS:

• Bedaquiline
• high performance liquid chromatography
• moxifloxacin
• pyrazinamide
• validation

Acknowledgements

Bhamini Rangnekar acknowledges an Honours studentship funded by the School of Pharmacy and MAMM a PhD scholarship from the University of Otago.

Additional information

Funding

This work is part of a research project supported by the Health Research Council of New Zealand (15/477).