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Significance: The production and characteristics of protonated small water clusters (PSWCs) were reported in this work, where in electrospray ionization (ESI) of pure water, the species obtained were singly charged molecular ions consisting of 2, 3, 4 or 5 water molecules attached to a hydrogen ion, [(H2O)n+H]+ , where n = 2, 3, 4 or 5. We proposed a new type of PSWCs structure: 2, 3, 4, 5 water molecules wrapped around a hydrogen ion which is located at the electrical and geometric center, forming a very stable molecular structure. Furthermore, biological tests of the PSWCs on mitochondrial function of intestinal epithelial cells and liver cells in mice showed the better therapeutic effect on inflammatory bowel diseases compared to that of the biologic agent Infliximab. Aim: To produce stable PSWCs as a new source of nutrient for health. Approach: The PSWCs were produced by electrospray technology, and identified by an electrospray mass spectrometer. The biological effects of the PSWCs on mitochondrial function of intestinal epithelial cells and liver cells in mice are carefully checked. Results: The production, characteristics and the biological effects of the PSWCs are studied, that we first generate the PSWCs by electrospray mechanism, which is proved to be charged (with positive charge). The obtained PSWCs are composed of 2, 3, 4, 5 water molecules plus a proton, respectively. The test of biological effects shows that the PSWCs can reduce radicals and improve the cell functions, indicating very important biological functions of the PSWCs. Conclusions: PSWCs was successfully produced by electrospray technology in our lab, the produced PSWCs were very stable under normal conditions without any obvious concentration changes of hydrogen ions in the last 3 years after produced, even the water is heated to 100 degrees Celsius. Furthermore, the effect of PSWCs on mitochondrial function of intestinal epithelial cells and liver cells in mice showed that the therapeutic effect of PSWCs on inflammatory bowel disease was better than that of the biologic agent Infliximab, where the higher the concentration of hydrogen ions, the better the therapeutic effect is.
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