Bacterial Disinfection Using Ozone Gas to Extent Spinach’s Shelf Life: The Effect of Dosage, Duration, and Spraying Frequency

Aprilia Ayuning Putri, Rais Salsa Muhammad, Eva Fathul Karamah


Spinach (Amaranthus tricolor) is one type of leafy vegetable famous as a nutrition source in Indonesia. However, it has short shelf life due to bacterial activity, which requires an alternative preservation solution besides drying. Ozone can be used as a disinfectant that is not harmful to humans and has been applied in food and agricultural product preservation. Ozone gas spraying reduces bacterial growth so that quality degradation due to decay can be slowed. In this research, spinach is preserved by spraying ozone gas to the extent of its shelf life. Spinach is ozonated with ozone gas at various dosages of 30.72 mg/hr, 48.60 mg/hr, and 80.16 mg/hr, with the spraying frequency of once, twice, and 3 times for 15 minutes. Ozone gas spraying duration is also variated: 2 minutes, 3 minutes, and 6 minutes. Then, the sample is stored for 7 days to see the change of its characteristic. The best quality during storage is shown by spinach treated with ozonation at a dosage of 48.60 mg/hr; 6 minutes; and the frequency of 3 times. In addition to variations in duration, dosage, and frequency, the same ozone exposure with different combination of duration-frequency and duration-dosage were also observed to evaluate the optimum ozone treatment for preserving spinach, and it was found that longest duration-least frequency and moderate duration-moderate dosage are preferred in maintaining the quality of spinach during storage.


Bacterial disinfection; ozone gas; preservation; shelf life; spinach.

Full Text:



H. Karaca, and Y.S. Velioglu,†Effects of ozone and chlorine washes and subsequent cold storage on microbiological quality and shelf life of fresh parsley leaves,†Food Science and Technology, vol. 127, pp. 1-9, 2020.

R. Yang, and G.B. Keding,†Nutritional contributions of important African indigenous vegetables,†In African indigenous vegetables in urban agriculture. C. M. Shackleton, M. W. Pasquini, and A. Drescher, Ed. London: Earthscan, 2009, pp. 105-143.

Alfaena, “Proyeksi Konsumsi Buah dan Sayur Utama di Indonesia Tahun 2018-2022,†B. Sc. Thesis, Institut Pertanian Bogor, Indonesia, 2018.

E. P. Wahyuni, “Mempelajari Karakteristik Pengeringan Bayam Hijau,†B. Sc. Thesis, Universitas Lampung, Indonesia, 2018.

A. A. Kader, Post-harvest technology of holticultural crops, 3rd ed., Oakland: University of California, 2002.

S. Shezi, L.S. Magwaza, A. Mditshwa, and S.Z. Tesfay,†Review. Changes in biochemistry of fresh produce in response to ozone postharvest treatment,†Scientia Horticulturae, vol. 269, pp.1-9, 2020.

R. Aslam, M.S. Alam, and P.A. Saeed, “Sanitization Potential of Ozone and Its Role in Postharvest Quality Management of Fruits and Vegetables,†Food Engineering Reviews, vol. 12, pp. 48–67, 2020.

E.F. Karamah and N. Wajdi, “Application of ozonated water to maintain the quality of chicken meat: effect of exposure time, temperature, and ozone concentration,†E3S Web of Conferences, vol. 67, no. 04044. 2018.

E.F. Karamah, N. Ismaningtyas, and A.P. Ilmiyah. “The effect of exposure time and water replacement in the application of ozonated water to maintain the quality of tunaâ€. IOP Conf. Ser.: Mater. Sci. Eng. Vol, 509, no. 012092, 2019.

A. Alexopoulos, S. Plessas, Y. Kourkoutas, C. Stefanis, S. Vavias, and C. Voidarou, “Experimental effect of ozone upon the microbial flora of commercially produced dairy fermented products,†International Journal of Food Microbiology, vol. 246, pp. 5–11, 2017.

F. Zhu. “Effect of ozone treatment on the quality of grain products,†Food Chemistry, vol. 264, pp. 358–366, 2018.

E.F. Karamah, F. Amalia, R. Ghaudenson, and S. Bismo, Disinfection of Escherichia coli Bacteria Using Combination of Ozonation and Hydrodynamic Cavitation Method with Venturi Injector,†International Journal on Advanced Science, Engineering and Information Technology, vol. 8, no. 3, pp. 811-817, 2018.

A. Baggio, · M. Marino, · N. Innocente, · M. Celotto, · and M. Maifreni, “Antimicrobial effect of oxidative technologies in food processing: an overview,†European Food Research and Technology, vol. 246, pp. 669–692, 2020.

R. Rachmawati, M.R. Defiani, and N. L. Suriani, “Pengaruh Suhu dan Lama Penyimpanan Terhadap Kandungan Vitamin C pada Cabai Rawit Putih,†Jurnal Biologi, vol. XIII, no. 2, pp. 36 – 40, 2009.

N. A. Buchmeier, S. J. Libby, Y. Xu, P. C. Loewen, J. Switala, and D. G. Guiney, “DNA repair is more important than catalase for Salmonella virulence in mice,†Journal of Clinical Investigation, vol. 95, no. 3, pp. 1047 – 1053, 1995.

N. K. Hunt and B. J. Marinas, “Inactivation of Escherichia coli with ozone: chemical and inactivation kinetics,†Water Research, vol. 33, no. 11, pp. 2633 – 2641, 1999.

S. Wani, J. Maker, J. Thompson, and J. Barnes, “Effect of Ozone Treatment on Inactivation of Eschericia coli and Listeria sp. on Spinach,†Agriculture, vol. 5, pp. 155-169, 2015.

D. Bermúdez-Aguirre, and G.V. Barbosa-Cánovas, “Disinfection of selected vegetables under nonthermal treatments: Chlorine, citric acid, ultraviolet light and ozone,†Food Control, vol. 29, pp. 82–90, 2013.

J.R. Junttila, S.I. Niemela and J. Hirn, “Minimum growth temperatures of listeria monocytogenes and non-haemolytic listeria,†Journal of Applied Bacteriology, vol. 65, pp. 321-327, 1988.

B. Maherani, · M. Harich, · S. Salmieri, · M. Lacroix, “Antibacterial Properties of Combined non-thermal treatments based on bioactive edible coating, ozonation, and gamma irradiation on ready-to-eat frozen green peppers: evaluation of their freshness and sensory qualities,†European Food Research and Technology, vol. 245, pp. 1095–1111, 2019.

A.M. Vettraino, V. Vinciguerra, G. Pacini, R. Forniti, V. Goffi, R. Botondi, “Gaseous Ozone as a Suitable Solution for Postharvest Chestnut Storage: Evaluation of Quality Parameter Trends,†Food and Bioprocess Technology, vol. 13, pp. 187–193, 2020.

L. Anderson, P. Koech, A. Plymale, E. Landorf, A. Konopka, F. Collart, M. Lipton, M. Romine, and A. Wright, “Live Cell Discovery of Microbial Vitamin Transport and Enzyme-Cofactor Interactions,†ACS Chemical Biology, vol. 11, no. 2, pp. 345-354, 2015.

A.J. Keutgen, and E. Pawelzik, “Influence of pre-harvest ozone exposure on quality of strawberry fruit under simulated retail condition,†Postharvest Biology and Technology, vol. 49, no. 1, pp. 10 – 18, 2008.

E. Porto, E.G.A. Filho, L.M.A. Silva, T.V. Fonteles, R.B.R. do Nascimento, F.A.N. Fernandesc, E. S. de Brito, and S. Rodrigues. “Ozone and plasma processing effect on green coconut water,†Food Research International, vol. 131, pp. 1-9, 2020.

L. Wang, X. Fan, K. Sokorai, and J. Sites, “Quality deterioration of grape tomato fruit during storage after treatments with gaseous ozone at conditions that significantly reduced populations of Salmonella,†Food Control, vol. 103, pp. 9–20. 2019.

G. Kaiser. (2019). Biology Libretexts: Factors that Influence Bacterial Growth. [Online]. Available:

T. Sopandi, and Wardah, Mikrobiologi Pangan – Teori dan Praktik. Yogyakarta: ANDI, 2016.



  • There are currently no refbacks.

Published by INSIGHT - Indonesian Society for Knowledge and Human Development