Effect of stabilizer nature and concentration time on storage of beet juice

Given the growing interest in natural beetroot betalains, the production of natural pigments of this class is a pressing issue. Concentrates produced from beetroot juice are in demand in the food, pharmaceutical, and medical industries. The properties specified by industry consumers require a stable system during long-term storage, preserving betanin, which has antioxidant activity. The stabilizing properties of citric acid (0.5%) and a mixture of citric (0.25%) and ascorbic (0.25%) acids on the optical properties of Bordeaux‑237 beet juice were studied during concentration and subsequent storage at 4±0.5°C using chemical analysis, electron microscopy, and Fourier transform infrared spectroscopy. The influence of the nature of the stabilizing agents on the kinetics of changes in betanin content and the composition of protein-carbohydrate components during beet juice concentration was determined. It was found that the nature of the stabilizer exerts a stabilizing effect on betanin, even with minor changes in beetroot juice pH during concentration. The increase in betanin content relative to the sample without the stabilizer (0.20%) reached 0.39% in the juice sample with citric acid and 0.35% in the juice sample stabilized with citric and ascorbic acids. The addition of citric acid significantly altered the concentration kinetics. As the 540 nm band maximum shifted to 535 nm, the 490 nm band decreased in intensity, and the weaker 455 nm band became more prominent. Betanin degradation in beetroot juice concentrates was determined to depend on concentration time. The change in spectral characteristics with concentration time is characterized by a delay of up to 10 minutes, followed by a sharp increase. The best system performance is achieved with a 15‑minute concentration at a temperature of 60±0.2°C and a pressure of 72±10 mbar. The minimum betanin loss in concentrates stabilized with citric acid after four months of refrigerated storage at 4±0.5°C was 5%.

1. Sangeeta, A., Barman, M., Mishra, P. (2025). Beverages and Concentrates. Chapter in a book: Fruits and Vegetables Technologies: Postharvest Processing and Packaging. Singapore: Springer Nature Singapore, 2025. https://doi.org/10.1007/978-981-96-8433-5_9

2. Saini, A., Hamid, Shams, R., Dash, K. K., Shaikh, A. M., Kovács, B. (2025). Anthocyanin extraction from black carrot: Health promoting properties and potential applications. Journal of Agriculture and Food Research, 19, Article 101533. https://doi.org/10.1016/j.jafr.2024.101533

3. Li, B., Lee, J. -Y., Luo, Y. (2023). Health benefits of astaxanthin and its encapsulation for improving bioavailability: A review. Journal of Agriculture and Food Research, 14, Article 100685. https://doi.org/10.1016/j.jafr.2023.100685

4. Deshmukh, G. P., Priyanka, Sindhav, R., Jose, N. (2018). Application of beetroot as natural coloring pigment and functional ingredient in dairy and food products. International Journal of Current Microbiology and Applied Sciences, 7(12), 2010–2016. https://doi.org/10.20546/ijcmas.2018.712.231

5. Saw, P. E., Lee, S., Jon, S. (2019). Naturally occurring bioactive compound-derived nanoparticles for biomedical applications. Advanced Therapeutics, 2(5), Article 1800146. https://doi.org/10.1002/adtp.201800146

6. Küçükgöz, K., Venema, K., Chamorro, F., Cassani, L., Donn, P., Prieto, M. A. et al. (2025). Unlocking the potential of fermented beetroot ketchup: Enhancing polyphenol recovery and gut microbiota interactions. Food Chemistry, 463 (Part 1), Article 141141. https://doi.org/10.1016/j.foodchem.2024.141141

7. Gengatharan, A., Dykes, G. A., Choo, W. S. (2015). Betalains: Natural plant pigments with potential application in functional foods. LWT‑Food Science and Technology, 64(2), 645–649. https://doi.org/10.1016/j.lwt.2015.06.052

8. Huang, D. (2018). Dietary antioxidants and health promotion. Antioxidants, 7(1), Article 9. https://doi.org/10.3390/antiox7010009

9. Sawicki, T., Bączek, N., Wiczkowski, W. (2016). Betalain profile, content and antioxidant capacity of red beetroot dependent on the genotype and root part. Journal of Functional Foods, 27, 249–261. https://doi.org/10.1016/j.jff.2016.09.004

10. Czylkowska, A., Rogalewicz, B., Szczesio, M., Raducka, A., Gobis, K., Szymański, P. et al. (2022). Antitumor activity against A549 cancer cells of three novel complexes supported by coating with silver nanoparticles. International Journal of Molecular Sciences, 23(6), Article 2980. https://doi.org/10.3390/ijms23062980

11. Bhat, S. A., Chandramohan, S., Krishna, G. S., Hiranmaya, C., Pajaniradje, S., Nair, A. S. et al. (2024). Betanin-encapsulated starch nanoparticles: Synthesis and cytotoxic effect on colon cancer. 3 Biotech, 14(10), Article 233. https://doi.org/10.1007/s13205-024-04078-4

12. Yin, Z., Yang, Y., Guo, T., Veeraraghavan, V. P., Wang, X. (2021). Potential chemotherapeutic effect of betalain against human non-small cell lung cancer through PI3K/Akt/mTOR signaling pathway. Environmental Toxicology, 36(6), 1011–1020. https://doi.org/10.1002/tox.23100

13. Farabegoli, F., Scarpa, E. S., Frati, A., Serafini, G., Papi, A., Spisni, E. et al. (2017). Betalains increase vitexin-2-O-xyloside cytotoxicity in CaCo-2 cancer cells. Food Chemistry, 218, 356–364. https://doi.org/10.1016/j.foodchem.2016.09.112

14. Singh, T., Pandey, V. K., Dash, K. K., Zanwar, S., Singh, R. (2023). Natural bio-colorant and pigments: Sources and applications in food processing. Journal of Agriculture and Food Research, 12, Article 100628. https://doi.org/10.1016/j.jafr.2023.100628

15. Küçükgöz, K., Venema, K., Trząskowska, M. (2024). Beetroot ketchup as a stable carrier of potential probiotic Lacticaseibacillus rhamnosus K3 and Lactobacillus johnsonii K4: A study on sensory attributes, storage viability, and in vitro gastrointestinal survival. Food and Bioproducts Processing, 148, 519–526. https://doi.org/10.1016/j.fbp.2024.10.004

16. Hadipour, E., Taleghani, A., Tayarani-Najaran, N., Tayarani-Najaran, Z. (2020). Biological effects of red beetroot and betalains: A review. Phytotherapy Research, 34(8), 1847–1867. https://doi.org/10.1002/ptr.6653

17. Madadi, E., Mazloum-Ravasan, S., Yu, J. S., Ha, J. W., Hamishehkar, H., Kim, K. H. (2020). Therapeutic application of betalains: A review. Plants, 9(9), Article 1219. https://doi.org/10.3390/plants9091219

18. Amjadi, S., Abbasi, M. M., Shokouhi, B., Ghorbani, M., Hamishehkar, H. (2019). Enhancement of therapeutic efficacy of betanin for diabetes treatment by liposomal nanocarriers. Journal of Functional Foods, 59, 119–128. https://doi.org/10.1016/j.jff.2019.05.015

19. Di, S., Yu, M., Guan, H., Zhou, Y. (2021). Neuroprotective effect of Betalain against AlCl3-induced Alzheimer’s disease in Sprague Dawley Rats via putative modulation of oxidative stress and nuclear factor kappa B (NF-κB) signaling pathway. Biomedicine and Pharmacotherapy, 137, Article 111369. https://doi.org/10.1016/j.biopha.2021.111369

20. Wang, J., Zhang, D., Cao, C., Yao, J. (2020). Betalain exerts a protective effect against glaucoma is majorly through the association of inflammatory cytokines. AMB Express, 10(1), Article 125. https://doi.org/10.1186/s13568-020-01062-y

21. Aztatzi-Rugerio, L., Granados-Balbuena, S. Y., Zainos-Cuapio, Y., Ocaranza-Sánchez, E., Rojas-López, M. (2019). Analysis of the degradation of betanin obtained from beetroot using Fourier transform infrared spectroscopy. Journal of Food Science and Technology, 56(8), 3677–3686. https://doi.org/10.1007/s13197-019-03826-2

22. Ahmadi, H., Nayeri, Z., Minuchehr, Z., Sabouni, F., Mohammadi, M. (2020). Betanin purification from red beetroots and evaluation of its anti-oxidant and anti-inflammatory activity on LPS-activated microglial cells. PLOS One, 15(5), Article e0233088. https://doi.org/10.1371/jounal.pone.0233088

23. Devadiga, D., Ahipa, T. N. (2020). Betanin: A Red-Violet Pigment — Chemistry and Applications. Chapter in a book: Chemistry and Technology of Natural and Synthetic Dyes and Pigments. IntechOpen, 2020.

24. Hu, T., Dai, T., He, X., Deng, L., Li, T., Sun, J. et al. (2023). Non-covalent interaction of complex plant protein and betanin: Mechanism of improving thermal stability of betanin. Food Hydrocolloids, 138, Article 108456. https://doi.org/10.1016/j.foodhyd.2023.108456

25. Racz, C. -P., Racz, L. Z., Floare, C. G., Tomoaia, G., Horovitz, O., Riga, S. (2023). Curcumin and whey protein concentrate binding: Thermodynamic and structural approach. Food Hydrocolloids, 139, Article 108547. https://doi.org/10.1016/j.foodhyd.2023.108547

26. Janiszewska-Turak, E., Walczak, M., Rybak, K., Pobiega, K., Gniewosz, M., Woźniak, Ł. et al. (2022). Influence of fermentation beetroot juice process on the physico-chemical properties of spray dried powder. Molecules, 27(3), Article 1008. https://doi.org/10.3390/molecules27031008

27. Murugesan, R., Orsat, V. (2012). Spray drying for the production of nutraceutical ingredients — A review. Food and Bioprocess Technology, 5(1), 3–14. https://doi.org/10.1007/s11947-011-0638-z

28. Koshani, R., Jafari, S. M. (2019). Ultrasound-assisted preparation of different nanocarriers loaded with food bioactive ingredients. Advances in Colloid and Interface Science, 270, 123–146. https://doi.org/10.1016/j.cis.2019.06.005

29. Martínez-Sánchez, A., Tarazona-Díaz, M. P., García-González, A., Gómez, P. A., Aguayo, E. (2016). Effect of high-pressure homogenization on different matrices of food supplements. Food Science and Technology International, 22(8), 708–719. https://doi.org/10.1177/1082013216642887

30. Nagai, R., Nagai, M., Shimasaki, S., Baynes, J. W., Fujiwara, Y. (2010). Citric acid inhibits development of cataracts, proteinuria and ketosis in streptozotocin (type 1) diabetic rats. Biochemical and Biophysical Research Communications, 393(1), 118–122. https://doi.org/10.1016/j.bbrc.2010.01.095

31. Yadikar, N., Ahmet, A., Zhu, J., Bao, X., Yang, X., Han, H. et al. (2022). Exploring the mechanism of citric acid for treating glucose metabolism disorder induced by hyperlipidemia. Journal of Food Biochemistry, 46(12), Article e14404. https://doi.org/10.1111/jfbc.14404

32. Miyagawa, Y., Fujita, H., Adachi, S. (2021). Kinetic analysis of thermal degradation of betanin at various pH values using deconvolution method. Food Chemistry, 361, Article 130165. https://doi.org/10.1016/j.foodchem.2021.130165

33. Trishitman, D., Negi, P. S., Rastogi, N. K. (2021). Concentration of beetroot juice colorant (betalains) by forward osmosis and its comparison with thermal processing. LWT, 145, Article 111522. https://doi.org/10.1016/j.lwt.2021.111522

34. Bazaria, B., Kumar, P. (2016). Compositional changes in functional attributes of vacuum concentrated beetroot juice. Journal of Food Processing and Preservation, 40(6), 1215–1222. https://doi.org/10.1111/jfpp.12705

35. Theba, T., Nayi, P., Ravani, A. (2024). Beetroot-based blended juice: Process development, physico-chemical analysis and optimization of novel health drink. Food Chemistry Advances, 4, Article 100607. https://doi.org/10.1016/j.focha.2024.100607

36. Monono, E. M., Wiesenborn, D. P., Vargas-Ramirez, J. M., Zhou, R. (2019). Preserving juice from industrial beets using organic acids. Transactions of the ASABE, 62(1), 177–185. https://doi.org/10.13031/trans.13051

37. 37.Thippeswamy, B., Joshi, A., Sethi, S., Dahuja, A., Kaur, C., Tomar, B. S. et al. (2022). Chemical additives for preserving the betalain pigment and antioxidant activity of red beetroot. Sugar Tech, 24(3), 890–899. https://doi.org/10.1007/s12355-021-01104-0

38. York, J. L., Roberts, M. P. (1976). The iron and subunit binding sites of hemerythrin. The role of histidine, tyrosine and tryptophan. Biochimica et Biophysica Acta (BBA)‑Protein Structure, 420(2), 265–278. https://doi.org/10.1016/0005-2795(76)90318-4

39. Pawade, S. S., Vilbaste, M., Siiman, A., Toom, L., Herodes, K., Leito, I. (2026). A quantitative approach to determine water and moisture content of different types of lignin using attenuated total reflectance Fourier transform infrared spectroscopy combined with partial least squares regression. Biofuels, Bioproducts and Biorefining, Early View. https://doi.org/10.1002/bbb.70124

40. Cengiz, M. F., Durak, M. Z. (2019). Rapid detection of sucrose adulteration in honey using Fourier transform infrared spectroscopy. Spectroscopy Letters, 52(5), 267–273. https://doi.org/10.1080/00387010.2019.1615957

41. Wang, J., Kliks, M. M., Jun, S., Jackson, M., Li, Q. X. (2010). Rapid analysis of glucose, fructose, sucrose, and maltose in honeys from different geographic regions using Fourier transform infrared spectroscopy and multivariate analysis. Journal of Food Science, 75(2), C208-C214. https://doi.org/10.1111/j.1750–3841.2009.01504.x

42. Barth, A., Zscherp, C. (2002). What vibrations tell about proteins. Quarterly Reviews of Biophysics, 35(4), 369–430. https://doi.org/10.1017/S0033583502003815

43. Barčot, O., Balarin, M., Gamulin, O., Ježek, D., Romac, P., Brnjas-Kraljević, J. (2007). Investigation of spermatozoa and seminal plasma by fourier transform infrared spectroscopy. Applied Spectroscopy, 61(3), 309–313. https://doi.org/10.1366/000370207780220804

44. Zannou, O., Oussou, K. F., Chabi, I. B., Odouaro, O. B. O., Deli, M. G. E. P., Goksen, G. et al. (2024). A comprehensive review of recent development in extraction and encapsulation techniques of betalains. Critical Reviews in Food Science and Nutrition, 64(30), 11263–11280. https://doi.org/10.1080/10408398.2023.2235695

45. Nemzer, B., Pietrzkowski, Z., Spórna, A., Stalica, P., Thresher, W., Michałowski, T. et al. (2011). Betalainic and nutritional profiles of pigment-enriched red beet root (Beta vulgaris L.) dried extracts. Food Chemistry, 127(1), 42–53. https://doi.org/10.1016/j.foodchem.2010.12.081