ISSN 2308-4057 (Печать),
ISSN 2310-9599 (Онлайн)

A phytochemical study of the clover growing in Kuzbass

In addition to studying bioactive organic compounds in plants, it is increasingly important to determine the biological role of elements in plants growing in environmentally unfavorable areas. One of such regions in Russia is Kuzbass with its intensively developing chemical, metallurgical, and coal mining sectors. In this study, we assessed the plant materials of red clover (Trifolium pratense L.), alsike clover (Trifolium hybridum L.), and white clover (Trifolium repens L.) collected from their natural populations in Kuzbass.
The qualitative and quantitative composition of heavy metals in the clover samples was determined voltammetrically. The contents of molybdenum and phosphorus were measured by the photocolorimetric method. Total nitrogen and protein were determined by the Kjeldahl method. Nickel, cobalt, and chromium were quantified by spectrophotometry.
We analyzed the plant materials of the clover samples for heavy metals and found that the content of lead was the least in red clover and the highest in alsike clover. Copper varied in a larger range and was minimal in red clover compared to that in alsike and white clover. Zinc was found at higher concentrations of in white and red clover compared to that in alsike clover. The levels of cadmium exceeded the maximum permissible concentrations in all the clover samples. We also revealed that the clover samples contained different amounts of various amino acids, including arginine, valine, lysine, glycine, aspartic acid, and alanine.
The plant materials of the clover species growing in Kuzbass can be used to improve the fertility of soil and nitrogen regime. However, the clover species should not be used in bulk feed for farm animals because of high concentrations of cadmium.
Ключевые слова
Clover, Trifolium pratense L., Trifolium hybridum L., Trifolium repens L., soil, microelements, amino acids, heavy metals, plants
This study was part of the Russian Science Foundation (RSF) project “Fundamentals of obtaining bioactive substances from Siberian medicinal plants and creating phytogenic feed additives with immunomodulatory effects on their basis” (Agreement No. 23-16-00113 dated May 15, 2023).
  1. Aslam M, Aslam A, Sheraz M, Ali B, Ulhassan Z, Najeeb U, et al. Lead toxicity in cereals: Mechanistic insight into toxicity, mode of action, and management. Frontiers in Plant Science. 2021;11.
  2. Chen G, Li J, Han H, Du R, Wang X. Physiological and molecular mechanisms of plant responses to copper stress. International Journal of Molecular Sciences. 2022;23(21).
  3. Asyakina LK, Dyshlyuk LS, Prosekov AYu. Reclamation of post-technological landscapes: International experience. Food Processing: Techniques and Technology. 2021;51(4):805–818. (In Russ.).
  4. Budantsev AL. Plant resources of Russia: Wild flowering plants, their composition and biological activity. Vol. 2. The families Actinidiaceae – Malvaceae, Euphorbiaceae – Haloragaceae. St. Petersburg, Moscow: KMK; 2009. 513 p. (In Russ.).
  5. Nizamutdinov ТI, Suleymanov AR, Morgun EN, Dinkelaker NV, Abakumov EV. Ecotoxicological analysis of fallow soils at the Yamal experimental agricultural station. Food Processing: Techniques and Technology. 2022;52(2):350–360.
  6. Titov AF, Talanova VV, Kaznina NM, Laidinen GF. Plant resistance to heavy metals. Petrozavodsk; 2007. 170 p. (In Russ.).
  7. Li X, Wang Z, Bai M, Chen Z, Gu G, Li X, et al. Effects of polystyrene microplastics on copper toxicity to the protozoan Euglena gracilis: Emphasis on different evaluation methods, photosynthesis, and metal accumulation. Environmental Science and Pollution Research. 2022;29:23461–23473.
  8. Pelikhovich YuV, Begday IV, Kharin KV, Tsesar TA. Heavy metal accumulation in medicinal plants and risk assessment. Science. Innovations. Technologies. 2020;(4):171–183. (In Russ.).
  9. Goncharuk EA, Zagoskina NV. Heavy metals, their phytotoxicity, and the role of phenolic antioxidants in plant stress responses with focus on cadmium: Review. Molecules. 2023;28(9).
  10. Capuozzo M, Santorsola M, Bocchetti M, Perri F, Cascella M, Granata V, et al. p53: From fundamental biology to clinical applications in cancer. Biology. 2022;11(9).
  11. Dʹyakova NA. The content of heavy metals and arsenic in the medicinal plants of the Voronezh region. Russia patent RU 2022620084. 2022.
  12. Ho L-H, Rode R, Siegel M, Reinhardt F, Neuhaus HE, Yvin J-C, et al. Potassium application boosts photosynthesis and sorbitol biosynthesis and accelerates cold acclimation of common plantain (Plantago major L.). Plants. 2020;9(10).
  13. Li Y, Yin M, Li L, Zheng J, Yuan X, Wen Y. Optimized potassium application rate increases foxtail millet grain yield by improving photosynthetic carbohydrate metabolism. Frontiers in Plant Science. 2022;13.
  14. Chen J, Zhang N-N, Pan Q, Lin X-Y, Shangguan Z, Zhang J-H, et al. Hydrogen sulphide alleviates iron deficiency by promoting iron availability and plant hormone levels in Glycine max seedlings. BMC Plant Biology. 2020;20.
  15. Dyshlyuk LS, Osintseva MA. Kozlova OV, Fotina NV, Prosekov AYu. Antiradical and oxidative stress release properties of Trifolium pratense L. extract. Journal of Experimental Biology and Agricultural Sciences. 2022;10(4):852–860.
  16. El'kina GYa. Content of amino acids in plants at different levels of lead in the soil. Agrohimia. 2023;(6):63–72. (In Russ.).
  17. Tabalenkona GN, Silina EV. Influence of habitat conditions on the content and composition of free amino acids of Plantago media L. leaves. Proceedings of Voronezh State University. Series: Chemistry. Biology. Pharmacy. 2023;(2):54–61. (In Russ.).
  18. Lezhnina MG, Khanina MA, Podolina EA, Rodin AP. The chemical composition of urban gravilate (Geum urbanum L.). The Prospects for Innovative Technologies in Medicine and Pharmacy: Proceedings of the 6th All-Russian Scientific and Practical Conference. Volume 2; 2019; Orekhovo-Zuevo. Orekhovo-Zuevo: State University of Humanities and Technology; 2019. p. 161–167. (In Russ.).
  19. Prosekov AYu, Ulrikh EV, Kozlova OV, Dyshlyuk LS. Study pectin as vegetable analog pharmaceutical gelatine. Modern Problems of Science and Education. 2014;(5). (In Russ.).
  20. Boyarskykh IG, Siromlya TI. Macro- and trace elements composition of some medicinal plants in the geochemically abnormal environment in the Altai Mountains (Russia). Rastitelnye Resursy. 2022;58(4):376–387.
  21. Yermokhin YuI, Bobrenko IA, Bobrenko EG. Microelement composition of agricultural plants in Siberia. Research and Scientific Electronic Journal of Omsk SAU. 2020;21(2).
  22. Kruglov DS, Prokusheva DL. The trace-element constituents of the most widespread plants of genus Artemisia. Chemistry of Plant Raw Material. 2022;(3):139–149. (In Russ.).
  23. Hilfiker A, Bovet L. Modulation of the content of amino acids in the plant. Russia patent RU 2799785C2. 2023.
  24. Saleh TA. Trends in the sample preparation and analysis of nanomaterials as environmental contaminants. Trends in Environmental Analytical Chemistry. 2020;28.
  25. Babich O, Sukhikh S, Pungin A, Asyakina L, Ivanova S, Prosekov A. Modern trends in the in vitro production and use of callus, suspension cells and root cultures of medicinal plants. Molecules. 2020;25(24).
  26. Babich O, Sukhikh S, Prosekov A, Asyakina L, Ivanova S. Medicinal plants to strengthen immunity during a pandemic. Pharmaceuticals. 2020;13(10).
  27. Polozhij AV, Vydrina SN, Kurbatskij VI, Nikiforova OD. Flora Sibiriae. Tomus 9. Fabaceae (Leguminosae). Novosibirsk: Nauka; 1994. 280 p. (In Russ.).
  28. Popova E. Distribution of heavy metals in plant communities of the West Siberian Arctic and Subarctic. E3S Web of Conferences. 2021;284.
  29. Sardans J, Peñuelas J. Potassium control of plant functions: Ecological and agricultural implications. Plants. 2021;10(2).
  30. Dyshlyuk L, Babich O, Prosekov A, Ivanova S, Pavsky V, Chaplygina T. The effect of postharvest ultraviolet irradiation on the content of antioxidant compounds and the activity of antioxidant enzymes in tomato. Heliyon. 2020;6(1).
  31. Dolganyuk V, Sukhikh S, Kalashnikova O, Ivanova S, Kashirskikh E, Prosekov A, et al. Food proteins: Potential resources. Sustainability. 2023;15(7).
  32. D'yakova, N. A. Peculiarities of heavy metal and arsenic trans-medium transition along the chain “soil – medicinal plant material – water etracts”. Humans and their Health. 2023;26(1):64–71. (In Russ.).
  33. Rubanka EV, Terletskaya VA, Zinchenko IN. The migration of heavy metals during the extraction of plant materials. Food Science and Technology. 2013;24(3):70–72. (In Russ.).
  34. Dmitrieva AI, Belashova OV, Ivanova SA, Prosekov AYu, Milenteva IS. Assessment of the content of heavy metals in medicinal plants of Genus trifolium from the growing area on the example of the Siberian federal district. International Journal of Pharmaceutical Research. 2020;12(3):1880–1893.
  35. Reut AA. The content of biologically active substances to the introduced members of the genus Hemerocallis L. News of FSVC. 2019;(1):93–96. (In Russ.).
  36. Faskhutdinova ER, Sukhikh AS, Le VM, Minina VI, Khelef MEA, Loseva AI. Effects of bioactive substances isolated from Siberian medicinal plants on the lifespan of Caenorhabditis elegans. Foods and Raw Materials. 2022;10(2):340–352.
  37. Kamanina IZ, Kaplina SP, Salikhova FS. The content of heavy metals in medicinal plants. Scientific Review. Biological Science. 2019;(1):29–34. (In Russ.).
  38. Kutlimurotova RKh, Pulatova LT. Study of the amino acid composition of Asarum europaeum L plants growing in Uzbekistan. Universum: Chemistry and Biology. 2021;86(8):27–30. (In Russ.).
  39. Babich OO, Milentyeva IS, Dyshlyuk LS, Ostapova EV, Altshuler OG. Structure and properties of antimicrobial peptides produced by antagonist microorganisms isolated from Siberian natural objects. Foods and Raw Materials. 2022;10(1):27–39.
  40. Dobrosmyslova IA, Sazanova AA, Semenov VG, Tuleubaev Zh, Yesimbekova ZT, Ziyaeva GK. Plant growth and biological productivity after processing with microelements. The Bulletin the National Academy of Sciences of the Republic of Kazakhstan. 2021;(1):74–80. (In Russ.).
  41. Prosekov AYu, Babich OO, Zaushintseva AV, Belashova OV, Milenteva IS, Asyakina LK, et al. Method for producing curd mass enriched with common skullcap and red clover concentrates. Russia patent RU 2753361C1. 2021.
  42. The State Pharmacopoeia of the Russian Federation. Vol. IV. Moscow; 2018. 1926 p. (In Russ.).
Как цитировать?
Belashova OV, Kozlova OV, Velichkovich NS, Fokina AD, Yustratov VP, Petrov AN. A phytochemical study of the clover growing in Kuzbass. Foods and Raw Materials. 2024;12(1):194–206. 
О журнале