Author(s): Nikulin A.V., Radus F.V., Panov A.V., Mironova E.V., Fedorina L.I., Martynov L. Y., Shatalov D. O., Potanina O. G., Ivanova A.V., Trashchenkova D.A., Dorovskikh E. A., Kovaleva T.Y.

Email(s): kovaleva_t_yu@staff.sechenov.ru

DOI: 10.52711/0974-360X.2026.00584   

Address: Nikulin A.V.1, Radus F.V.1, Panov A.V.1, Mironova E.V.1, Fedorina L.I.1, Martynov L. Y.1, Shatalov D. O.1, Potanina O. G.2, Ivanova A.V.3, Trashchenkova D.A.3, Dorovskikh E. A.3, Kovaleva T.Y.1,3*
1MIREA - Russian Technological University, 78 Vernadsky Ave, 119454, Russian Federation.
2Federal State Budget Educational Institution of Higher Education M.V.Lomonosov Moscow State University (Lomonosov MSU), Leninskie Gory, 1, Moscow, 119991, Russian Federation.
3Sechenov First Moscow State Medical University (Sechenov University), 8 Trubetskaya St., bldg. 2, Moscow, 119991, Russian Federation.
*Corresponding Author

Published In:   Volume - 19,      Issue - 9,     Year - 2026


ABSTRACT:
Red clover (Trifolium pratense L.) and zigzag clover (Trifolium medium L.), family Fabaceae – legumes, are widespread in the world flora, have valuable medicinal properties. The Trifolium pratense L. herb monographs are included in the State Pharmacopoeia of the Russian Federation XV edition and USP 36/NF31, Trifolium medium L. is currently being studied. The overlapping habitat of both Trifolium species and similar external appearance make it important to differentiate both plants during the harvesting and the herbal drug when identification determining. To conduct a comparative macro- and microscopic analysis of red clover (Trifolium pratense L.) and zigzag clover (Trifolium medium L.) herb to establish the specific characteristics of external and anatomical diagnostic features. Red clover (Trifolium pratense L.) and zigzag clover (Trifolium medium L.) herb samples were collected in Moscow region of the Russian Federation in summer 2024. The analysis of microscopic structures was carried out on a Leica DM1000 light microscope (Germany) with a 10x/20 eyepiece and 10x/0.25 and 40x/0.65 objectives. A comparative analysis of the macroscopic and microscopic features of the red clover (Trifolium pratense L.) and zigzag clover (Trifolium medium L.) herb was carried out. The stems, leaves, inflorescences were examined. External features were determined, their importance for differentiating species features both in nature and in determining identification of medicinal plant raw material were revealed. The size and frequency of occurrence per 1 mm2 of the main anatomical-diagnostic features of all parts of the herb were established.


Cite this article:
Nikulin A.V., Radus F.V., Panov A.V., Mironova E.V., Fedorina L.I., Martynov L. Y., Shatalov D. O., Potanina O. G., Ivanova A.V., Trashchenkova D.A., Dorovskikh E. A., Kovaleva T.Y.. Comparative Study of Macroscopic and Microscopic Features of Red Clover (Trifolium pratense L.) and Zigzag Clover (Trifolium medium L.). Research Journal Pharmacy and Technology. 2026;19(9):4173-1. doi: 10.52711/0974-360X.2026.00584

Cite(Electronic):
Nikulin A.V., Radus F.V., Panov A.V., Mironova E.V., Fedorina L.I., Martynov L. Y., Shatalov D. O., Potanina O. G., Ivanova A.V., Trashchenkova D.A., Dorovskikh E. A., Kovaleva T.Y.. Comparative Study of Macroscopic and Microscopic Features of Red Clover (Trifolium pratense L.) and Zigzag Clover (Trifolium medium L.). Research Journal Pharmacy and Technology. 2026;19(9):4173-1. doi: 10.52711/0974-360X.2026.00584   Available on: https://rjptonline.org/AbstractView.aspx?PID=2026-19-9-30


REFERENCES: 
1.    Ahmad S, Zeb A. Phytochemical profile and pharmacological properties of Trifolium repens. J Basic Clin Physiol Pharmacol. 2021; 32(1). doi:10.1515/jbcpp-2020-0015
2.    Kshnikatkina AN, Grishin GYe, Gorbunov MV. Formation of legume-rhizobium symbiosis of clover pannonian variety Anik depending on methods of cultivation. Niva Povolzhya. 2016; (3): 39–48. [in Russian].
3.    Efremov AP. Medicinal plants and fungi of Central Russia. Moscow: Fitogon XXI; 2018. 504 p. [in Russian].
4.    Syutkina AI. Clover grass as a promising source of medicinal plant raw materials. Molodoy Uchenyi. 2015; 11(91): 719–21. [in Russian].
5.    Dluhošová J, Ištvánek J, Nedělník J, Řepková J. Red clover (Trifolium pratense) and zigzag clover (T. medium) – a picture of genomic similarities and differences. Front Plant Sci. 2018; 9: 724.
6.    Ates E, Tenikecier HS. Content of isoflavones and macroelements in five clover species (Trifolium spp.) grown on pasture: the influence of a slope aspect. J Elementol. 2022; 27(2): 379–92. doi:10.5601/jelem.2022.27.2.2240
7.    Maciejewska-Turska M, Zgorka G. In-depth phytochemical and biological studies on potential AChE inhibitors in red and zigzag clover dry extracts using RP-LC–PDA–ESI-QToF/MS-MS and TLC-bioautography. Food Chem. 2021; 375: 131846. doi:10.1016/j.foodchem.2021.131846
8.    Dluhošová J, Řepková J, Jakešová H, Nedělník J. Impact of interspecific hybridization of T. pratense × T. medium and backcrossing on genetic variability of progeny. Genet Plant Breed. 2016;52(1):125–31.
9.    Chaudhary N, Tripathi S. A review on chemical and biological activity of Trifolium pratense. PharmaTutor. 2014; 2(3): 93–101.
10.    Bajaj M, Bahri S, Sinha R. Phytoestrogens of Trifolium pratense L. as therapeutics: a review. South Asian J Exp Biol. 2023; 13(4): 252–62. doi:10.38150/sajeb.13(4).p252-262
11.    Lagari V, Levis S. Phytoestrogens for menopausal bone loss and climacteric symptoms. J Steroid Biochem Mol Biol. 2014; 139: 294–301. doi:10.1016/j.jsbmb.2012.12.002
12.    Myers S, Vigar V. Effects of a standardized extract of Trifolium pratense (Promensil) at a dosage of 80 mg in the treatment of menopausal hot flushes: a systematic review and meta-analysis. Phytomedicine. 2017; 24: 141–7. doi:10.1016/j.phymed.2016.12.003
13.    Zgorka G, Maciejewska-Turska M, Makuch-Kocka A, Plech T. In vitro evaluation of the antioxidant activity and chemopreventive potential in human breast cancer cell lines of the standardized extract obtained from the aerial parts of zigzag clover (Trifolium medium L.). Pharmaceuticals. 2022; 15(6): 699. doi:10.3390/ph15060699
14.    Arjun S. An Overview on Phytoestrogen based antihypertensive agent for their potential Pharmacological Mechanism. Research Journal of Pharmaceutical Dosage Forms and Technology. 2023; 15(3): 211-4.
15.    Ahsas G, Jeetendra KG, Debapriya G. Phytoestrogens are emerging medicine in Prevention and Management of Cognitive deficits in Postmenopausal Women. Research J. Pharm. and Tech. 2021; 14(1): 513-515.
16.    Harsha VT, Shalini BR, Sachin SP, Amol KR, Arvind RU. Soyabean as a Neutraceutical a Phytopharmacological: Review. Research J. Pharmacognosy and Phytochemistry. 2012; 4(2): 112-118.
17.    Vlaisavljević S, Kaurinović B, Popović M, Vasiljević S. Profile of phenolic compounds in Trifolium pratense L. extracts at different growth stages and their biological activities. Int J Food Prop. 2017; 20(3): 3090–101. doi:10.1080/10942912.2016.1273235
18.    Lemežienė N, Padarauskas A, Butkutė B, Ceseviciene J, Taujenis L, Norkevičienė E. The concentration of isoflavones in red clover (Trifolium pratense L.) at flowering stage. Zemdirbyste. 2015; 102(4): 443–8. doi:10.13080/z-a.2015.102.057
19.    Les F, Cásedas G, López V. Bioactivity of medicinal plants and extracts. Biology (Basel). 2021; 10(7): 634. doi:10.3390/biology10070634
20.    Zhang H, Zhao J, Shang H, Guo Y, Chen S. Extraction, purification, hypoglycemic and antioxidant activities of red clover (Trifolium pratense L.) polysaccharides. Int J Biol Macromol. 2020; 148: 750–60. doi:10.1016/j.ijbiomac.2020.01.194
21.    Kaurinović B, Popović M, Vlaisavljević S, Schwartsova H, Vojinović-Miloradov M. Antioxidant profile of Trifolium pratense L. Molecules. 2012; 17(9): 11156–72. doi:10.3390/molecules170911156
22.    Noor MN, Talal AA, Ibrahim SA. Isolation of Isoflavones from Iraqi Trifolium pretense. Research Journal of Pharmacy and Technology. 2022; 15(10):4692-6.
23.    Haflin Soraya Hutagalung, Aldy Safruddin Rambe, Tri Widyawati, Mustafa Mahmud Amin, Endang Mutiawati Rahayuningsih, Kiking Ritarwan, Puji Pinta Omar Sinurat, Sabri Ibrahim. Red clover (Trifolium pratense) Extract Ability Enhances Motor Coordination and Spatial Memory in Parkinson Murine Model. Research Journal of Pharmacy and Technology. 2025; 18(10): 4949-4.
24.    Ramasamy A, Aiyalu R. Fingerprint Analysis of Herbal Raw Materials available in Market Belongs to Combretacea And Fabaceae family by HPTLC technique using antioxidant markers. Asian Journal of Pharmacy and Technology. 2021; 11(2):130-4.
25.    Saranya Sh, Mythili S. A Comparative Study on the Biological Activities of Medicinal Plants Leaf Extracts Belongs to the Family Fabaceae. Research J. Pharm. and Tech. 2019; 12(3): 1129-1136.
26.    Hisham A. Abbas. Antibacterial, Anti-swarming and Antibiofilm Activities of Local Egyptian Clover Honey Against Proteus Mirabilis Isolated from Diabetic Foot Infection. Asian J. Pharm. Res. 3(3): July-Sept. 2013; Page 114-117.
27.    Butkutė B, Lemežienė N, Padarauskas A, Norkevičienė E, Taujenis L. Chemical composition of zigzag clover (Trifolium medium L.). In: Breeding Grasses and Protein Crops in the Era of Genomics. Cham: Springer; 2018. p. 83–7. doi:10.1007/978-3-319-89578-9_15
28.    Gharibzahedi SMT, Jafari SM. The importance of minerals in human nutrition: bioavailability, food fortification, processing effects and nanoencapsulation. Trends Food Sci Technol. 2017;62:119–32. doi:10.1016/j.tifs.2017.02.017
29.    Zgórka G, Maciejewska-Turska M. Cultivation and utilization of red clover (Trifolium pratense L.). In: Medicinal Plants: Domestication, Biotechnology and Regional Importance. Cham: Springer; 2021. p. 315–38.
30.    Dyshlyuk L, Fedorova A, Loseva A, Eremeeva N. Callus cultures of Thymus vulgaris and Trifolium pratense as a source of geroprotectors. Food Process Tech Technol. 2021;51(2):423–32. [in Russian] doi:10.21603/2074-9414-2021-2-423-432
31.    Manzoureh R, Farahpour MR. Topical administration of hydroethanolic extract of Trifolium pratense (red clover) accelerates wound healing by apoptosis and re-epithelialization. Biotech Histochem. 2021;96(4):276–86. doi:10.1080/10520295.2020.1797875
32.    Kubasova ED, Krylov IA, Kubasov RV, Nezgovorov DV, Korobitsyn AP, Groshilin VS, et al. Perspectives for the use of plant raw materials of the clover meadow (Trifolium pratense L.) in pharmacy. Hum Ecol. 2024;31(3). [in Russian] doi:10.17816/humeco634190
33.    Novikov VS, Shustov EB. The role of minerals and microelements in maintaining human health. Herald Educ Sci Dev RANS. 2017; 21(3): 5–16. [in Russian].
34.    Goldfein MD, Adaev ON, Timush LG, Zaikov GE, Yaroshevskaya HM. The role of chemical elements and their compounds in nature and in human life processes. Part 1. Chemicals in ecology, microelementoses and general issues of food safety. Herald Technol Univ. 2015; 18(16): 296–300. [in Russian].
35.    Vikrant A, Raneev Th. Plant Saponins –A Recent Update. Asian J. Research Chem. 6(9): September 2013; Page 871-876.
36.    Anjali S, Patel F, Preeti Sh. In-vitro Cytotoxic Activity of Plant Saponin Extracts on Breast Cancer Cell-Line. Res. J. Pharmacognosy and Phytochem. 2017; 9(1): 17-22.
37.    Nguyen TSh, Ditkovskaya ZR, Generalova YuE, Kaukhova IE, Sorokin VV. Determination of the elemental composition of red clover grass (Trifolium pratense L.). Probl Biol Med Pharm Chem. 2020;23(2):51–5. [in Russian] doi:10.29296/25877313-2020-02-08
38.    Gounden T, Moodley R, Jonnalagadda SB. Elemental analysis and nutritional value of edible Trifolium (clover) species. Environ Sci Health B. 2018; 53(8): 487–92. doi:10.1080/03601234.2018.1462923
39.    Dyshlyuk L, Osintseva M, Kozlova O, Fotina N, Prosekov A. Antiradical and oxidative stress release properties of Trifolium pratense L. extract. J Exp Biol Agric Sci. 2022; 10(4): 852–60. doi:10.18006/2022.10(4).852.860
40.    State Pharmacopoeia of the Russian Federation. XV edition [Internet]. Moscow: Ministry of Health of the Russian Federation; [cited 2024 Sep 19]. Available from: https://pharmacopoeia.regmed.ru/pharmacopoeia/izdanie-15/
41.    The United States Pharmacopeia: USP 36/NF 31. Baltimore (MD): United States Pharmacopeial Convention; 2013. p. 1572–4.Interactive Agricultural Ecological Atlas of Russia and Neighboring Countries [Internet]. [cited 2024 Sep 19]. Available from: https://agroatlas.ru/en/index.htmlublication). Access mode: https://agroatlas.ru/en/index.html

Recomonded Articles:

Research Journal of Pharmacy and Technology (RJPT) is an international, peer-reviewed, multidisciplinary journal.... Read more >>>

RNI: CHHENG00387/33/1/2008-TC                     
DOI: 10.52711/0974-360X 

1.3
2021CiteScore
 
56th percentile
Powered by  Scopus


SCImago Journal & Country Rank

Journal Policies & Information


Recent Articles




Tags


Not Available