Rifampicin reacts with cupric ion to produce rifampicin-Cu(II) chelate (2:1) at pH 6.5, which can be extracted with methylisobutylketone (MIBK). Therefore, rifampicin can be quantitatively, determined by measuring the quantity of Cu(II) in the organic phase by atomic absorption spectrophotometry. Because higher absorbance ratio was obtained in pH 6.0~7.0, buffer solution at pH 6.5 was used in this measurement. Linear relationship was found between absorbance and concentration in the range of 1.0~4.0${ imes}10^{-4}M$. This method might be applicable to the determination of rifampicin in the preparations.