The acid dissociation constants of the hydroxyl groups in tetrasodium 25,26,27,28-tetrahydroxycalix[4]arene-5,11,17,23-tetrasulfonate (CALX-S4) were determined at 25-degrees-C by potentiometric and spectrophotometric titration methods. The first acid dissociation constant(pK(a1)) was found to be 3.26 +/- 0.02 (mu = 0.1 with KNO3), which demonstrated a remarkable pK(a) shift due to intramolecular hydrogen-bonding interactions among the hydroxyl groups in CALX-S4. On the other hand, considerably weak acidities were observed on the residual three hydroxyl groups of CALX-S4 (pK(a2) = 11.8 +/- 0.3 (mu = 0.1 with KNO3), 11.3 +/- 0.3 (mu = 2.0 with KCl), pK(a3) = 12.8 +/- 0.3 (mu = 2.0 with KCl), and pK(a4) = ca. 14 (mu = 2.0 with KCl)).