To resolve the differentiation problem of acetone and acrolein in the analysis of carbonyls by high-performance liquid chromatography (HPLC), we investigated the optimum analytical conditions for their separation. Carbonyl compounds were collected by 2,4-dinitrophenylhydrazine (DNPH)-coated cartridges. We examined the influence of three experimental variables: temperature (25, 30, 40, 50 and 60 ${^{circ}C}$), flow rate (1.0 and 1.2 mL/min), and relative mobile phase composition (among acetonitrile, water and tetrahydrofuran). The experimental results revealed the optimum analytical condition of a flow rate of 1.2 mL/min, temperature of 32 ${^{circ}C}$ and mobile phase composition of acetonitrile: water: tetrahydrofuran = 34 : 52.8 : 13.2. The analysis of indoor air composition indicated that acrolein and acetone comprised 11% and 42% of all aldehydes, respectively.