In general, the distillation parameters, such as the number of theoretical plate (NTP) and the height equivalent to a theoretical plate (HETP), can be obtained from the operation at the steady state. However, it is time-consuming to achieve the steady state especially in the case of isotope separation. In this paper, with the purpose of simultaneous separation of isotopes of nitrogen and oxygen by NO distillation, we tried to determine the distillation parameters by an analytical method through the transient-state operation. It was confirmed that the results from the analysis were in good agreement with those observed for the operation at the steady state. Enrichment of the isotopes was carried out using a distillation column with a height of 1 m and inside diameter of 12 mm. The dependence of HETP on liquid flow rate was measured by the proposed method. The obtained HETP values were from 2 to 4 cm. The operation time of about 5 h was found to be long enough to determine the distillation parameters.