Validation of a simple isocratic HPLC-UV method for rifampicin and isoniazid quantification in human plasma

LUCIANI GIACOBBE, CAROLINA; GUZMAN, MARIA LAURA; MANZO, R.H.; MARIA E OLIVERA ME

Journal of Applied Pharmaceutical Science

Open Science Publishers LLP

Año: 2018 vol. 8 p. 93 - 93

simple and selective reversed phase HPLC-UV method for rifampicin and isoniazid quantification in human plasmawas developed and validated. The method consisted of drug extraction with trichloroacetic acid and organic solvent followed by derivatization of isoniazid. Using an isocratic mode, rifampicin was analyzed on a C18 (250 × 4.6 mm, 5µm) column at 339 nm, while isoniazid was analyzed on a C8 (250 × 4.6 mm, 5 µm) column at 273 nm. All validation parameters fulfilled the FDA requirements, as the method was accurate (bias% < 10.26), precise (CV% < 10.39) and linear from 0.31 to 37.80 μg/mL of rifampicin and 0.89 to 71.36 μg/mL of isoniazid. The samples remained stable during the usual processing and analysis times and also during the two freeze/thaw cycles. The recovery of both analytes was reproducible (CV% < 11.2) in the range of 97.3-99.6% of rifampicin and 89.8-96.6% of isoniazid.The low volume of plasma necessary for the quantification of the samples (750 µL in total)