Convenient methods for ingestion of molecular hydrogen: drinking, injection, and inhalation
© Kurokawa et al. 2015
Received: 3 August 2015
Accepted: 9 October 2015
Published: 26 October 2015
Molecular hydrogen (H2) is clinically administered; however, in some hospitals, H2 is given to patients without consideration of its safe use. In the present study, we prepared convenient and safe devices for the drinking of super-saturated H2 water, for intravenous drip infusion of H2-rich saline, and for the inhalation of H2 gas. In order to provide useful information for researchers using these devices, the changes in H2 concentration were studied. Our experimental results should contribute to the advance of non-clinical and clinical research in H2 medicine.
KeywordsHydrogen water Hydrogen-rich saline Hydrogen gas
Molecular hydrogen (H2) is a medical gas with beneficial effects on oxidative stress , inflammation , apoptosis , lipid metabolism , and signaling pathways . More than 280 articles, including 24 articles on clinical studies, have demonstrated that H2 ameliorates the pathological conditions in numerous human diseases  or disease models in animals , since Ohsawa et al. reported that H2 could be used in antioxidant therapy .
H2 is clinically administered through the oral intake of H2 water [9–12], intravenous drip infusion of H2-rich saline [12–15], or inhalation of air with 2-4 % H2 gas . However, in some hospitals, H2 is given to patients by intravenous drip infusion and/or inhalation without consideration of its safe use. We have developed and provided various devices for the ingestion of H2 to solve this problem. Furthermore, the beneficial effects of H2 using our devices have been reported in 7 human diseases [9–16].
In the present study, we prepared convenient and safe devices for drinking super-saturated H2 water, for intravenous drip infusion of H2-rich saline, and for the inhalation of H2 gas. We examined the changes in H2 concentrations in these devices in order to provide useful information for researchers. Our experimental results reported in this article should contribute to the advance of non-clinical and clinical research in H2 medicine.
Preparation of super-saturated H2 water for drinking
Similarly, H2 water was obtained by the use of non-woven fabric. As shown in Fig. 1c and d, the non-woven fabric containing H2-generating agent (0.65 g) was first inserted into an acrylic resin tube, and 0.5 mL of water was added. The tube was tightly closed with a cap attached to a check valve, and inserted into a pressure-resistant PET bottle filled with water. H2 generated in the tube was transferred to the bottle through the valve. In about 5 min at room temperature, the agent started a reaction in the wet fabric. The H2 gas produced was emitted into the water through the check valve attached to the acrylic resin tube. During the reaction, the PET bottle was gradually pressurized to approximately 6 atmospheric pressures due to the generation of H2 gas. After 24 h, the H2 gas was dissolved by shaking the bottle for about 30 s.
Preparation of H2-rich saline for injection
As shown in Fig. 2c, non-woven fabric containing the H2-generating agent was moistened with a small amount of water, and then both a drip infusion bag and the non-woven fabric were wrapped with aluminum foil under reduced pressure. The water reacted with the agent in the non-woven fabric to generate H2, and the H2 gas permeating through the polyethylene film in the bag dissolved into the solution.
Preparation of H2-containing gas for inhalation
Measurement of H2 concentration
The concentration of H2 gas in the water was measured using the methylene blue platinum colloid reagent-based titration method, as described previously , and verified using an electrochemical gas sensor (model DHD1-1, DKK-TOA Corp., Tokyo, Japan). On the other hand, the concentration of H2 in the air was measured using an H2 gas sensor (FIS Inc., Hyogo, Japan).
The concentration of H2 gas in the water or air is presented as ppm (mg/L, weight/volume) or % (volume/volume), respectively. Most of the experimental data are expressed as mean ± standard deviation (mean ± SD) of more than three individual measurements. However, in the examination of H2-rich saline, the H2 concentration is expressed as an individual measurement to examine the differences between each bag and plastic vessel. The statistical significance was assessed by Student’s paired or unpaired t-test for single comparisons or by one-way analysis of variance (ANOVA) followed by Fisher’s LSD test for multiple comparisons. A p value of less than 0.05 was considered to be statistically significant.
H2 concentration of super-saturated H2 water prepared by Method I
H2 concentration of super-saturated H2 water prepared by Method II
H2 concentration of H2-rich saline prepared by Method III
Details of drip infusion bag, dialysis fluid bag, and injection ampoule used in the experiment
5 % Glucose injection
Solulact (Lactate ringer sol.)
Isotonic sodium chloride sol.
Otsuka normal saline
Hartman’s sol. pH 8 (Lactate ringer sol.)
5 % Glucose injection (for animals)
7 % Sodium hydrogen carbonate sol. (for animals)
Otsuka normal saline
Otsuka normal saline
Isotonic sodium chloride sol.
Isotonic sodium chloride sol.
H2 concentration of H2-rich saline prepared by Method IV
H2 concentration of gas introduced by inhaler
We examined the H2 gas concentration for up to 3 h after starting use of the inhaler, because stability of the gas concentration is required in order to examine the performance of the gas inhaler. The H2 gas concentration in the inhaler was 2.91 ± 0.08 % after 0.5 min, and a H2 gas concentration of approximately 3 % was maintained for 3 h. There was no significant difference among of the time points after starting (Fig. 7b). These results demonstrate that the H2 gas could be supplied stably for 3 h using this inhaler.
In summary, we prepared two types of super-saturated H2 water (7 or 10 ppm) for drinking. The concentrations in these waters were maintained for 7 days without opening, but the waters should be drunk within 2 h of the cap being opened. We also prepared two types of H2-rich saline for injection. Although intravenous drip injection with the H2-rich saline should be completed within 1 h, H2 concentrations in the saline prepared by aluminum foil (Method IV) were maintained for 12 months without opening. Moreover, we prepared H2-containing gas for inhalation. The gas was controlled under the detonation limit of the mixture of H2 gas and air, and the gas could be supplied stably for 3 h. In a recent study, we examined the H2 concentration in rat tissue following administration of H2 via various routes . We demonstrated that H2 concentrations in the tissues depend on the H2 concentration of the administered water or gas, and that the specific uptake of H2 in the tissues is due to the difference in administration route . The present results suggest the importance in choosing the more efficient route of H2 treatment for each disease or tissue . Therefore, we believe that the super-saturated H2 water (10 ppm) prepared by Method II, the H2-rich saline prepared by Method IV, and the H2 gas prepared by our method are convenient and safe preparatory methods. The present results should contribute to the advance of non-clinical and clinical research in H2 medicine.
- H2 :
The authors are grateful to Dr. Yoshio Okamoto for his excellent advice in the writing of this manuscript.
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- Li J, Wang C, Zhang JH, Cai JM, Cao YP, Sun XJ. Hydrogen-rich saline improves memory function in a rat model of amyloid-beta-induced Alzheimer’s disease by reduction of oxidative stress. Brain Res. 2010;1328:152–61.View ArticlePubMedGoogle Scholar
- Xie K, Yu Y, Zhang Z, Liu W, Pei Y, Xiang L, et al. Hydrogen gas improves survival rate and organ damage in zymosan-induced generalized inflammation model. Shock. 2010;34(5):495–501.View ArticlePubMedGoogle Scholar
- Yang Y, Li B, Liu C, Chuai Y, Lei J, Gao F, et al. Hydrogen-rich saline protects immunocytes from radiation-induced apoptosis. Med Sci Monit. 2012;18(4):BR144–148.PubMed CentralView ArticlePubMedGoogle Scholar
- Song G, Li M, Sang H, Zhang L, Li X, Yao S, et al. Hydrogen-rich water decrease serum low-density lipoprotein cholesterol levels and improves high-density lipoprotein function in patients with potential metabolic syndrome. J Lipid Res. 2013;54(2):1884–93.PubMed CentralView ArticlePubMedGoogle Scholar
- Itoh T, Fujita Y, Itoh M, Masuda A, Ohno K, Ichihara M, et al. Molecular hydrogen suppresses FcepsilonRI-mediated signal transduction and prevents degranulation of mast cells. Biochem Biophys Res Commun. 2009;389(4):651–8.View ArticlePubMedGoogle Scholar
- Nakao A, Toyoda Y, Sharma P, Evans M, Guthrie N. Effectiveness of hydrogen rich water on antioxidant status of subjects with potential metabolic syndrome: an open label pilot study. J Clin Biochem Nutr. 2010;46(2):140–9.PubMed CentralView ArticlePubMedGoogle Scholar
- Yuan L, Chen X, Qian L, Shen J, Cai J. Administration of hydrogen-rich saline in mice with allogeneic hematopoietic stem-cell transplantation. Med Sci Monit. 2015;21:749–54.PubMed CentralView ArticlePubMedGoogle Scholar
- Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, et al. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med. 2007;13(6):688–94.View ArticlePubMedGoogle Scholar
- Yoritaka A, Takanashi M, Hirayama M, Nakahara T, Ohta S, Hattori N. Pilot study of H2 therapy in Parkinson’s disease. A randomized double-blind placebo-controlled trial. Mov Disord. 2013;28(6):836–9.View ArticlePubMedGoogle Scholar
- Ishibashi T, Sato B, Rikitake M, Seo T, Kurokawa R, Hara Y, et al. Consumption of water containing a high concentration of molecular hydrogen reduces oxidative stress and disease activity in patients with rheumatoid arthritis: an open-label pilot study. Med Gas Res. 2012;2(1):27.PubMed CentralView ArticlePubMedGoogle Scholar
- Sakai T, Sato B, Hara K, Hara Y, Naritomi Y, Koyanagi S, et al. Consumption of water containing over 3.5 mg of dissolved hydrogen could improve vascular endothelial function. Vasc Health Risk Manage. 2014;10:591–7.Google Scholar
- Ishibashi T, Ichikawa M, Sato B, Shibata S, Hara Y, Naritomi Y, et al. Improvement of psoriasis-associated arthritis and skin lesions by treatment with molecular hydrogen: A report of three cases. Mol Med Rep. 2015;12(2):2757–64.PubMedGoogle Scholar
- Ono H, Nishijima Y, Adachi N, Tachibana S, Chitoku S, Mukaihara S, et al. Improved brain MRI indices in the acute brain stem infarct sites treated with hydroxyl radical scavengers, Edaravone and hydrogen, as compared to Edaravone alone. A non-controlled study. Med Gas Res. 2011;1(1):12.PubMed CentralView ArticlePubMedGoogle Scholar
- Ono H, Nishijima Y, Adachi N, Sakamoto M, Kudo Y, Nakazawa J, et al. Hydrogen (H2) treatment for acute erythymatous skin disease. A report of 4 patients with safety data and a non-controlled feasibility study with H2 concentration measurement on two volunteers. Med Gas Res. 2012;2(1):14.PubMed CentralView ArticlePubMedGoogle Scholar
- Ishibashi T, Sato B, Shibata S, Sakai T, Hara Y, Naritomi Y, et al. Therapeutic efficacy of infused molecular hydrogen in saline on rheumatoid arthritis: A randomized, double-blind placebo-controlled pilot study. Int Immunopharmacol. 2014;21(1):468–73.View ArticlePubMedGoogle Scholar
- Terawaki H, Zhu WJ, Matsuyama Y, Terada T, Takahashi Y, Sakurai K, et al. Effect of hydrogen (H2)-enriched solution on the albumin redox of hemodialysis patients. Hemodial Int. 2014;18(2):459–66.View ArticlePubMedGoogle Scholar
- Seo T, Kurokawa R, Sato B. A convenient method for determining the concentration of hydrogen in water: Use of methylene blue with colloidal platinum. Med Gas Res. 2012;2(1):1.PubMed CentralView ArticlePubMedGoogle Scholar
- Liu C, Kurokawa R, Fujino M, Hirano S, Sato B, Li XK. Estimation of the hydrogen concentration in rat tissue using an airtight tube following the administration of hydrogen via various routes. Sci Rep. 2014;4:5485.PubMed CentralPubMedGoogle Scholar