Objective: To find out the optimal separating conditions and wavelength program for measurement of 16 polycyclic aromatic hydrocarbons in house dust by high-performance liquid chromatography (HPLC) with fluorescence detector. Methods: Vacuum cleaner was used for gathering the house dust,and acetone was added and extraction was performed in a ultrasonic bath. The supernatant was concentrated and then added methyl cyanide to 0.1 ml. With methyl cyanide and water as mobile phase,PAHs were efficiently separated by gradient elution and detected by fluorescence detector and wavelength programming. Results: Except for acenaphthylene,15 PAHs had highly efficient separation within 28 min and showed good linearity with 0.5-50 ng/ml range. The relative standard deviations were 1.35%-5.89%,and the recoveries were 62.45%-110.36%. Conclusion: HPLC with fluorescence detection and wavelength programming could be used for detection of PAHs in house dust because of its quickness,simplicity,sensitivity and reproducibility.