Few pharmacokinetic data of acamprosate were available in Chinese population and no medication is approved for alcohol dependence in China. 1. Investigate the pharmacokinetic properties of acamprosate calcium in healthy Chinese male volunteers on single- and multiple-dose administration. 2. Compare the bioequivalence of two formulations of acamprosate calcium tablets both under fasting and fed conditions. This open-label, randomized study included 3 stages. In each stage, a 2-way crossover bioequivalence study was conducted to study the pharmacokinetic properties and bioequivalence of acamprosate calcium tablets on multiple dosing after standardized meals, single dosing under fasting conditions and fed conditions, respectively. The washout period between each treatment in a stage and between each stage was 1week. Plasma acamprosate calcium was quantified by a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Tolerability was evaluated by monitoring adverse events, physical examinations, 12-lead ECG, and laboratory tests. Totally, 36 male subjects were enrolled in the study and all of them completed the whole 3 study stages. Main pharmacokinetic parameters of test and reference formulations were as follows: multiple dosing, Tmax 9.94±6.59 and 9.47±5.47h, Cmax 435.74±348.10 and 346.54±155.66ng·mL(-1), AUC0-t 8600.52±5264.77 and 9315.10±6820.03ng·mL(-1)·h, AUC0-∞ 8845.38±5838.18 and 9669.24±7326.53ng·mL(-1)·h, t1/2 10.06±8.83 and 9.87±10.35h; single dosing under fasting conditions, Tmax 7.29±4.87 and 6.57±1.85h, Cmax 247.85±110.05 and 244.64±132.43ng·mL(-1), AUC0-t 3385.41±1418.92 and 3496.24±1767.29ng·mL(-1)·h, AUC0-∞ 3781.53±1556.96 and 3829.56±1981.25ng·mL(-1)·h, t1/2 13.07±17.24 and 10.26±7.78h; single dosing under fed conditions, Tmax 17.72±9.42 and 19.50±9.84h, Cmax 183.90±74.52 and 168.14±60.67ng·mL(-1), AUC0-t 3181.71±1368.24 and 3575.11±1416.39ng·mL(-1)·h, AUC0-∞3442.39±2002.53 and 3624.44±1418.12ng·mL(-1)·h, t1/2 8.76±12.28 and 6.67±4.84h, respectively. In all three stages, 90% CIs for the test/reference ratio of AUC0-t and AUC0-∞ were located within 80%-125%, 90% CI for Cmax was within 70%-143%. Similar pharmacokinetic results of acamprosate calcium tablets in healthy Chinese volunteers were found as those in Caucasic population. In all three stages, the two formulations met the regulatory criteria for bioequivalence. Chictr.org identifier: ChiCTR-TTRCC-14004853. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.