ANTIBACTERIAL, IN VITRO CYTOTOXIC, AND ANTIOXIDANT ACTIVITIES OF ELECTROLYZED OXIDIZING/REDUCING WATER
DOI:
https://doi.org/10.22159/ijap.2019.v11s5.T0043Keywords:
Antibacterial activity, Antioxidant, Anticancer, Electrolyzed water, A healthy waterAbstract
Objective: Electrolyzed oxidizing/reducing water is popular as health beneficial water in Indonesia. In this study, we examined the level of antibacterial,
anticancer, and antioxidant activity of the electrolyzed water.
Methods: The efficacy of electrolyzed water produced by Enagic® at six level pH (2.5, 6.0, 7.0, 8.5, 9.0, and 9.5) was investigated. Antibacterial
activity was evaluated using a macrodilution method. The anticancer activity was performed against human breast cancer (T47D) cell lines using
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Moreover, the antioxidant activity was determined using antioxidant model,
2,2-diphenyl-1picrylhydrazyl radical scavenging activity.
Results: The results show that electrolyzed water exhibited antibacterial activity against Propionibacterium acnes and Staphylococcus epidermidis.
Among six level pH, electrolyzed water at pH 2.5 showed the highest antibacterial activity. The in vitro cytotoxic activity of electrolyzed water showed
potential moderate cytotoxicity. The activity tends to be higher in alkaline electrolyzed water. However, the electrolyzed water showed free radical
scavenging activity.
Conclusion: Electrolyzed water that marked in Indonesia has some potential health benefits. The activity is dependent on pH.
Downloads
Download data is not yet available.
References
1. Ignatov I, Gluhchev G, Karadzhov S, Miloshev G, Ivanov N, Mosin O.
Preparation of electrochemically activated water solutions (catholyte/
anolyte) and studying their physical-chemical properties. J Health Med
Nurs 2015;13:64-78.
2. Rahman SM, Khan I, Oh DH. Electrolyzed water as a novel sanitizer in
the food industry: Current trends and future perspectives. Compr Rev
Food Sci Food Saf 2016;15:471-90.
3. Sharma RR, Demirci A. Treatment of Escherichia coli O157:H7
inoculated alfalfa seeds and sprouts with electrolyzed oxidizing water.
Int J Food Microbiol 2003;86:231-7.
4. Bari ML, Sabina Y, Isobe S, Uemura T, Isshiki K. Effectiveness
of electrolyzed acidic water in killing Escherichia coli O157:H7,
Salmonella enteritidis, and Listeria monocytogenes on the surfaces of
tomatoes. J Food Prot 2003;66:542-8.
5. Al Haq MI, Sugiyama J, Isobe S. Review applications of electrolyzed
Fig. 2: In vitro cytotoxic activity of electrolyzed water against
T47D cell lines
Fig. 3: Phase contrast images of T47D cells after incubation with
100% of electrolyzed waters at pH 2.5, 6.0, and 9 for 24 h (×20)
Table 3: Antioxidant activity of electrolyzed water, using DPPH
free radical scavenging method
Treatment EC50±SD (%) EC20±SD (%)
pH 2.5 227.4±9.7 102.5±8.0
pH 6.0 293.0±34.4 130.3±16.5
pH 7.0 607.4±28.7 219.6±13.1
pH 8.5 195.8±4.5 63.3±2.3
pH 9.0 177.4±1.6 63.2±4.0
pH 9.5 167.6±2.8 56.0±3.2
Ascorbic acid 9.3±0.4 (?g/mL) 3.4±0.3 (?g/mL)
DPPH: 2,2?diphenyl?1picrylhydrazyl, EC50: 50% efficient concentration
water in agriculture and food industries. Food Sci Technol Res
2005;11:135-50.
6. Kim MJ, Kim HK. Anti-diabetic effects of electrolyzed reduced
water in streptozotocin-induced and genetic diabetic mice. Life Sci
2006;79:2288-92.
7. Ye J, Li Y, Hamasaki T, Nakamichi N, Komatsu T, Kashiwagi T, et al.
Inhibitory effect of electrolyzed reduced water on tumor angiogenesis.
Biol Pharm Bull 2008;31:19-26.
8. Tsai CF, Hsu YW, Chen WK, Ho YC, Lu FJ. Enhanced induction of
mitochondrial damage and apoptosis in human leukemia HL-60 cells
due to electrolyzed-reduced water and glutathione. Biosci Biotechnol
Biochem 2009;73:280-7.
9. Saitoh Y, Harata Y, Mizuhashi F, Nakajima M, Miwa N. Biological
safety of neutral-pH hydrogen-enriched electrolyzed water upon
mutagenicity, genotoxicity and subchronic oral toxicity. Toxicol Ind
Health 2010;26:203-16.
10. Shirahata S, Kabayama S, Nakano M, Miura T, Kusumoto K, Gotoh M,
et al. Electrolyzed-reduced water scavenges active oxygen species and
protects DNA from oxidative damage. Biochem Biophys Res Commun
1997;234:269-74.
11. Shirahata S, Hamasaki T, Teruya K. Advanced research on the health
benefit of reduced water. Trends Food Sci Technol 2012;23:124-31.
12. Moorman E, Montazeri N, Jaykus LA. Efficacy of neutral electrolyzed
water for inactivation of human norovirus. Appl Environ Microbiol
2017;83:e00653-17.
13. Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method
to evaluate antioxidant activity. Lebenson Wiss Technol 1995;28:25-30.
14. Kedare SB, Singh RP. Genesis and development of DPPH method of
antioxidant assay. J Food Sci Technol 2011;48:412-22.
15. Nakagawara S, Goto T, Nara M, Ozawa Y, Hotta K, Arata Y.
Spectroscopic characterization and the pH dependence of bactericidal
activity of the aqueous chlorine solution. Anal Sci 1998;14:691-8.
16. Venkitanarayanan KS, Ezeike GO, Hung YC, Doyle MP. Efficacy of
electrolyzed oxidizing water for inactivating Escherichia coli O157:H7,
Salmonella enteritidis, and Listeria monocytogenes. Appl Environ
Microbiol 1999;65:4276-9.
17. McPherson LL. Understanding ORP’s in the disinfection process.
Water Eng Manage 1993;140:29-31.
18. Marriott NG, Gravani RB. Principles of Food Sanitation. 5th ed.
New York: Springer; 2006.
19. Korting HC, Lukacs A, Vogt N, Urban J, Ehret W, Ruckdeschel G,
et al. Influence of the pH-value on the growth of Staphylococcus
epidermidis, Staphylococcus aureus and Propionibacterium acnes in
continuous culture. Zentralbl Hyg Umweltmed 1992;193:78-90.
20. Sandvik EL, McLeod BR, Parker AE, Stewart PS. Direct electric current
treatment under physiologic saline conditions kills Staphylococcus
epidermidis biofilms via electrolytic generation of hypochlorous acid.
PLoS One 2013;8:e55118.
21. O’Connor PM, Jackman J, Bae I, Myers TG, Fan S, Mutoh M, et al.
Characterization of the p53 tumor suppressor pathway in cell lines of
the national cancer institute anticancer drug screen and correlations
with the growth-inhibitory potency of 123 anticancer agents. Cancer
Res 1997;57:4285-300.
22. Blois MS. Antioxidant determinations by the use of a stable free radical.
Nature 1958;181:1199-200.
23. Hamasaki T, Harada G, Nakamichi N, Kabayama S, Teruya K, Fugetsu B,
et al. Electrochemically reduced water exerts superior reactive oxygen
species scavenging activity in HT1080 cells than the equivalent level of
hydrogen-dissolved water. PLoS One 2017;12:e0171192.
24. Pisoschi AM, Negulescu GP. Methods for total antioxidant activity
determination: A review. Biochem Anal Biochem 2011;1:1000106.
Preparation of electrochemically activated water solutions (catholyte/
anolyte) and studying their physical-chemical properties. J Health Med
Nurs 2015;13:64-78.
2. Rahman SM, Khan I, Oh DH. Electrolyzed water as a novel sanitizer in
the food industry: Current trends and future perspectives. Compr Rev
Food Sci Food Saf 2016;15:471-90.
3. Sharma RR, Demirci A. Treatment of Escherichia coli O157:H7
inoculated alfalfa seeds and sprouts with electrolyzed oxidizing water.
Int J Food Microbiol 2003;86:231-7.
4. Bari ML, Sabina Y, Isobe S, Uemura T, Isshiki K. Effectiveness
of electrolyzed acidic water in killing Escherichia coli O157:H7,
Salmonella enteritidis, and Listeria monocytogenes on the surfaces of
tomatoes. J Food Prot 2003;66:542-8.
5. Al Haq MI, Sugiyama J, Isobe S. Review applications of electrolyzed
Fig. 2: In vitro cytotoxic activity of electrolyzed water against
T47D cell lines
Fig. 3: Phase contrast images of T47D cells after incubation with
100% of electrolyzed waters at pH 2.5, 6.0, and 9 for 24 h (×20)
Table 3: Antioxidant activity of electrolyzed water, using DPPH
free radical scavenging method
Treatment EC50±SD (%) EC20±SD (%)
pH 2.5 227.4±9.7 102.5±8.0
pH 6.0 293.0±34.4 130.3±16.5
pH 7.0 607.4±28.7 219.6±13.1
pH 8.5 195.8±4.5 63.3±2.3
pH 9.0 177.4±1.6 63.2±4.0
pH 9.5 167.6±2.8 56.0±3.2
Ascorbic acid 9.3±0.4 (?g/mL) 3.4±0.3 (?g/mL)
DPPH: 2,2?diphenyl?1picrylhydrazyl, EC50: 50% efficient concentration
water in agriculture and food industries. Food Sci Technol Res
2005;11:135-50.
6. Kim MJ, Kim HK. Anti-diabetic effects of electrolyzed reduced
water in streptozotocin-induced and genetic diabetic mice. Life Sci
2006;79:2288-92.
7. Ye J, Li Y, Hamasaki T, Nakamichi N, Komatsu T, Kashiwagi T, et al.
Inhibitory effect of electrolyzed reduced water on tumor angiogenesis.
Biol Pharm Bull 2008;31:19-26.
8. Tsai CF, Hsu YW, Chen WK, Ho YC, Lu FJ. Enhanced induction of
mitochondrial damage and apoptosis in human leukemia HL-60 cells
due to electrolyzed-reduced water and glutathione. Biosci Biotechnol
Biochem 2009;73:280-7.
9. Saitoh Y, Harata Y, Mizuhashi F, Nakajima M, Miwa N. Biological
safety of neutral-pH hydrogen-enriched electrolyzed water upon
mutagenicity, genotoxicity and subchronic oral toxicity. Toxicol Ind
Health 2010;26:203-16.
10. Shirahata S, Kabayama S, Nakano M, Miura T, Kusumoto K, Gotoh M,
et al. Electrolyzed-reduced water scavenges active oxygen species and
protects DNA from oxidative damage. Biochem Biophys Res Commun
1997;234:269-74.
11. Shirahata S, Hamasaki T, Teruya K. Advanced research on the health
benefit of reduced water. Trends Food Sci Technol 2012;23:124-31.
12. Moorman E, Montazeri N, Jaykus LA. Efficacy of neutral electrolyzed
water for inactivation of human norovirus. Appl Environ Microbiol
2017;83:e00653-17.
13. Brand-Williams W, Cuvelier ME, Berset C. Use of a free radical method
to evaluate antioxidant activity. Lebenson Wiss Technol 1995;28:25-30.
14. Kedare SB, Singh RP. Genesis and development of DPPH method of
antioxidant assay. J Food Sci Technol 2011;48:412-22.
15. Nakagawara S, Goto T, Nara M, Ozawa Y, Hotta K, Arata Y.
Spectroscopic characterization and the pH dependence of bactericidal
activity of the aqueous chlorine solution. Anal Sci 1998;14:691-8.
16. Venkitanarayanan KS, Ezeike GO, Hung YC, Doyle MP. Efficacy of
electrolyzed oxidizing water for inactivating Escherichia coli O157:H7,
Salmonella enteritidis, and Listeria monocytogenes. Appl Environ
Microbiol 1999;65:4276-9.
17. McPherson LL. Understanding ORP’s in the disinfection process.
Water Eng Manage 1993;140:29-31.
18. Marriott NG, Gravani RB. Principles of Food Sanitation. 5th ed.
New York: Springer; 2006.
19. Korting HC, Lukacs A, Vogt N, Urban J, Ehret W, Ruckdeschel G,
et al. Influence of the pH-value on the growth of Staphylococcus
epidermidis, Staphylococcus aureus and Propionibacterium acnes in
continuous culture. Zentralbl Hyg Umweltmed 1992;193:78-90.
20. Sandvik EL, McLeod BR, Parker AE, Stewart PS. Direct electric current
treatment under physiologic saline conditions kills Staphylococcus
epidermidis biofilms via electrolytic generation of hypochlorous acid.
PLoS One 2013;8:e55118.
21. O’Connor PM, Jackman J, Bae I, Myers TG, Fan S, Mutoh M, et al.
Characterization of the p53 tumor suppressor pathway in cell lines of
the national cancer institute anticancer drug screen and correlations
with the growth-inhibitory potency of 123 anticancer agents. Cancer
Res 1997;57:4285-300.
22. Blois MS. Antioxidant determinations by the use of a stable free radical.
Nature 1958;181:1199-200.
23. Hamasaki T, Harada G, Nakamichi N, Kabayama S, Teruya K, Fugetsu B,
et al. Electrochemically reduced water exerts superior reactive oxygen
species scavenging activity in HT1080 cells than the equivalent level of
hydrogen-dissolved water. PLoS One 2017;12:e0171192.
24. Pisoschi AM, Negulescu GP. Methods for total antioxidant activity
determination: A review. Biochem Anal Biochem 2011;1:1000106.
Published
15-09-2019
How to Cite
DJALIL, A. D., SUNDHANI, E., WAHYUNINGRUM, R., HARTANTI, D., LESTIOWATI, N., PANGESTU, P., SETIAJI, R., & SUWANDRI, S. (2019). ANTIBACTERIAL, IN VITRO CYTOTOXIC, AND ANTIOXIDANT ACTIVITIES OF ELECTROLYZED OXIDIZING/REDUCING WATER. International Journal of Applied Pharmaceutics, 11(5), 40–43. https://doi.org/10.22159/ijap.2019.v11s5.T0043
Issue
Section
Original Article(s)
