A simple analytical method for the determination of p-nitrobenzene-azo-naphthol was proposed by a sequential perturbation with different amounts of p-nitrobenzene-azo-naphthol on an oscillating chemical system. The method involves a Cu(II)-catalysed oscillating reaction between hydrogen peroxide and sodium thiocyanate in alkaline medium with the aid of continuous-flow stirred tank reactor (CSTR). A good linear relationship between the changes, oscillation amplitude or/and period, and the concentration of p-nitrobenzene-azo-naphthol was obtained. The use of analyte pulse perturbation technique provides a possibility of sequential determination in a same oscillating system, due to a new steady state that reappeared rapidly after each perturbation. The calibration curve fits a linear equation very well when the concentration of p-nitrobenzene-azo-naphthol ranging from 5.2×10−7 to 3.3×10−3M. Influence of temperature, injection point, flow rate and reactants variables on this system were investigated in detail.