Gastric acid measurement is useful in assessing the effectiveness of antisecretory drugs, however, the conventional tests involve invasive nasogastric intubation. Orally administered 13 C-labeled calcium carbonate (Ca 13 CO 3 ) reacts with gastric acid to produce 13 C-labeled carbon dioxide ( 13 CO 2 ), which is then excreted in the breath. The objective of this study was to evaluate the suitability of Ca 13 CO 3 breath test for estimating gastric acid secretion in human noninvasively. First, the Ca 13 CO 3 breath test and the measurement of pooled gastric acid under a fasting condition were performed in 6 healthy volunteers to evaluate the correlation between the two parameters. Next, endoscopic gastric acid collection and the Ca 13 CO 3 breath test were performed on different days after pentagastrin injection in 20 subjects to evaluate the correlation between the tests and the reproducibility. Finally, the same studies were repeated in 4 subjects before and after 1-week rabeprazole, a proton pump inhibitor, administration. The maximum 13 CO 2 concentration (Cmax) correlated very well with the amount of pooled gastric acid (r = 0.95), suggesting that Ca 13 CO 3 breath test values well reflected the fasting intragastric acidity. The 13 CO 2 concentration after pentagastrin injection correlated well with pentagastrin-stimulated maximal acid output (r = 0.79 at 20 min). The reproducibility of the Ca 13 CO 3 breath test under pentagastrin-stimulation was good (coefficient of variation = 0.11). Rabeprazole administration markedly reduced the values of the Ca 13 CO 3 breath test, suggesting that it can sensitively assess the efficacy of rabeprazole. The Ca 13 CO 3 breath test can potentially be a useful method for non-invasive estimation for gastric acid secretion in human.