We have synthesized the 1,3-alternate 25,27-dioctyloxy-26,28-bis-[3- aminopropyloxy]-calix[4]arene and then immobilized onto γ- chloropropylsilica gel (CPS). The high-performance liquid chromatographic behavior of some aromatic hydrocarbons, phenolic compounds, aromatic amines and drug compounds was studied on this 1,3-alternate-calix[4]arene-bonded silica gel stationary phase (CIMS). The effect of organic modifier content and pH of the mobile phase on retention and selectivity of these compounds were investigated. According to chromatographic data, it can be concluded that the selectivity of CIMS for analytes ascribes to various interactions between CIMS and the analytes, such as hydrophobic interaction, hydrogen bonding interaction, π-π interaction and inclusion interaction. © 2010 Elsevier B.V. All rights reserved.