The sensitive analysis of 90Sr with accelerator mass spectrometry (AMS) was developed to advance environmental radiology. One advantage of AMS is the ability to analyze environmental samples with 90Sr/88Sr atomic ratios of 10-14 in following a simple chemical separation. Three different IAEA samples with known 90Sr concentrations (moss-soil, animal bone, Syrian soil: 1 g each) were analyzed to assess the validity of the chemical separation and the AMS measurement. The 90Sr measurements were conducted on the AMS system VERA combined with the Ion Laser InterAction Mass Spectrometry (ILIAMS) setup at the University of Vienna, which has excellent isobaric separation performance. The isobaric interference of 90Zr in the 90Sr AMS was first largely removed by chemical separation. The separation factor of Zr in two-step column chromatography with Sr resin and anion exchange resin was 106. The 90Zr remaining in the sample was effectively suppressed by ILIAMS. This procedure achieved a limit of detection <0.1 mBq in the 90Sr AMS, which is lower than typical β-ray detection. The agreement between AMS measurements and nominal values for the 90Sr concentrations of IAEA samples indicated that the new highly-sensitive 90Sr analysis in the environmental samples with AMS is reliable.