Direct inlet probe (DIP) was used as an introduction and a pre-separation step for atmospheric pressure photoionization time-of-flight ion mobility spectrometry (APPI-TOF-IMS) for the first time. IMS is an analytical technique used to separate and identify ionized molecules in the gas phase and under atmospheric pressure based on their mobility. The utilization of DIP prior to IMS gives the possibility to introduce the analytes into the gas phase and provides an additional separation based on their vapor pressure. The proof-of-principle study was done on example of eight polycyclic aromatic hydrocarbons (PAHs) with the ring number from 2 to 5, namely naphthalene, fluorene, anthracene, phenanthrene, pyrene, fluoranthene, benzo[a]pyrene, and benzo[k]fluoranthene. All these compounds are included in EPA priority pollutant list. Moreover, benzo[a]pyrene and benzo[k]fluoranthene are marked by EPA as probably carcinogen compounds and also included into SCF and EU lists. To increase the sensitivity of DIP-APPI-IMS the analysis was performed using a dopant assisted ionization method (benzene, 74 mg L−1 in N2). It was found that the heating rate of the interface plays a crucial role for the whole analytical procedure. To prove the ability of this method to analyze PAHs in the mixture, the mixtures containing up to five PAHs were analyzed. The LODs for the analyzed compounds obtained with DIP-APPI-IMS were found to be in the tens- or hundreds-of-microgram-per-liter range. The obtained results are promising enough to ensure the potential of DIP as an introduction and a pre-separation step for ion mobility based methods.