Fine dust has emerged as seriously hazardrous particles, which can cause respiratory and cardiovascular diseases. Thus, we have developed a natural complex extract consisted of Codonopsis lanceolata (Siebold & Zucc.) Trautv., Platycodon grandiflorum A. De Candolle, and Glycyrrhiza uralensis Fischer in the ratio of 3 : 5 : 2, which have a beneficial effect on respiratory system. In this study, simultaneous quantitative analysis of marker compounds, lobetyolin, platycodin D, and glycyrrhizic acid in the natural complex extract was developed and validated with high performance liquid chromatography-photodiode array detector. The marker compounds were shown in a large linearity with a correlation coefficient (R2) of 1.000. The limit of detection (LOD) of lobetyolin, platycodin D and glycyrrhizic acid were 5.40 ㎍/mL, 3.94 ㎍/mL and 5.86 ㎍/mL, respectively. The limit of quantification (LOQ) of lobetyolin, platycodin D and glycyrrhizic acid were 16.36 ㎍/mL, 11.94 ㎍/mL and 17.75 ㎍/mL, respectively. The content analysis revealed that the amounts of lobetyolin, platycodin D, and glycyrrhizic acid were 0.039±0.013 mg/g, 0.337±0.048 mg/g, and 1.171±0.003 mg/g, respectively, in the natural complex extract. Therefore, these results could be used as basic data for the standardization and quality control of the natural complex extract.