Evaluation of antimicrobial activity of dosage forms based on Michurin's chokeberry leaves

Introduction. Recently, researchers have shown great interest in studying the phytochemical composition of Michurin's aronia leaves (Aronia × mitschurinii A.K. Skvortsov & Maitul) due to the significant content of such valuable biologcally active substances (BAS) as flavonoids, leucoanthocyanins and tannins of the condensed group. Considering the high potential astringent, capillary-protective and anti-inflammatory activity of the main target groups of the BAS of the studied raw materials, in the framework of the present study we have analysed the antimicrobial activity of aqueous-alcoholic liquid extracts of the medicinal forms from Michurin's chokeberry leaves. In our recent studies, we have also established the biocidal effect of the decoction in vitro at a dilution of 1 : 10 on the test system of infusoria Parametium caudatum under the damaging effect of sodium chloride solution in the test "Functional load".

Aim. The purpose of the work was to determine the antimicrobial activity of aqueous-alcoholic liquid extraction dosage forms obtained from the leaves of chokeberry Michurin, in order to assess the prospects for creating medicinal herbal preparations (MHRP).

Materials and methods. The object of the study were liquid extraction dosage forms, tincture (1 : 5) and liquid extract (1 : 1), obtained from dried leaves of Michurin's chokeberry using 60 % ethanol. The antimicrobial activity of the samples was determined by the double serial dilution method. Staphylococcus aureus, Bacillus cereus, Candida albicans, Escherichia coli, Pseudomonas aeruginosa strains were used as test cultures. The prediction of the main types of pharmacological activity by in silico method was carried out for the pre-identified components of the leaf polyphenolic complex – catechin, rutin and leucocyanidin – using PassOnline and Phyto4Health platform. The medicinal forms obtained were standardised in accordance with the requirements for tinctures and extracts of the State Pharmacopoeia XV edition.

Results and discussion. The in silico screening of the main target groups of the BAS of the studied raw materials confirmed their high potential astringent and anti-inflammatory activity. The BAS complex of the liquid extract of Michurin's chokeberry leaves in comparison with the tincture shows antimicrobial activity against Pseudomonas aeruginosa up to the 4^th dilution and antifungal activity against Candida albicans up to the 6th dilution. In relation to such pathogenic microorganisms as Bacillus cereus and Staphylococcus aureus both tested dosage forms showed antimicrobial activity up to the 6^th–7^th dilution. The dosage form with a wider spectrum of antimicrobial activity is the liquid extract. In addition, the obtained dosage forms were standardized according to the requirements of the State Pharmacopoeia of the Russian Federation.

Conclusion. The results of the evaluation of the antimicrobial activity of the obtained alcoholic dosage forms based on Michurin's chokeberry leaves (tincture and liquid extract), confirmed the prospects of creating herbal medicines, e.g. for external use in the complex therapy and prevention of diseases of the oral cavity in the form of rinses or applications.

1. Brezhneva T. A., Nedoluzhko E. I., Logvinova E. E, Gudkova A. A., Slivkin A. I. The study of biologically active substances in the leaves of black chokeberry. Vestnik VGU. Seriya: Khimiya. Biologiya. Farmatsiya. 2018;2:306–311. (In Russ.)

2. Nedoluzhko E. I., Brezhneva T. A., Logvinova E. E. et al. Study of biologically active substances of chokeberry leaves. In: University science: a look into the future. Collection of scientific papers based on the materials of the International Scientific Conference dedicated to the 83rd anniversary of Kursk State Medical University. 2018. P. 74–77. (In Russ.)

3. Pugacheva O. V., Sviridova O. L., Brezhneva T. A., Slivkin A. I. Validation of a method for the quantification of tannins in black chokeberry leaves. Proceedings of Voronezh State University. Series: Chemistry. Biology. Pharmacy. 2022;1:98–104. (In Russ.)

4. Pugacheva O. V., Brezhneva T. A., Slivkin A. I. Determination of tannins in the leaves of chokeberry by various analytical methods. In: Collection of proceedings of the seventh scientific conference "Modern trends in the development of health-saving technologies". 2019. P. 292–298. (In Russ.)

5. Pugacheva O. V., Brezhneva T. A., Slivkin A. I. Determination of tannins in chokeberry leaves to select the timing of raw material procurement. In: "Modern trends in the development of health-saving technologies", collection of materials from the IX International Scientific Conference of Young People scientists. 2021. P. 345–350. (In Russ.)

6. Pugacheva O. V., Brezhneva T. A., Slivkin A. I. Determination of flavonoids in chokeberry leaves by TLC. In: “Ways and forms of improving pharmaceutical education. Current issues in the development and research of new drugs”, collection of proceedings of the 8th International Scientific and Methodological Conference. 2022. P. 429–433. (In Russ.)

7. Pugacheva O. V., Trineeva O. V., Panova K. E. Quantitative determination of anthocyanins in leaves of chokeberry Michurin. In: Ways and forms of improving pharmaceutical education. Current issues of development and research new medicines. Collection of proceedings of the 9th International Scientific and Methodological Conference. Voronezh. 2023. P. 356–359. (In Russ.)

8. Pugacheva O. V., Brezhneva T. A., Slivkin A. I. Development and validation of a method for the quantification of flavonoids in aronia Michurina leaves. Proceedings of Voronezh State University. Series: Chemistry. Biology. Pharmacy. 2023;3:92–99. (In Russ.)

9. Pugacheva O. V., Trineeva O. V., Panova K. E. Validation of a method for the quantification of antocianins in aronia Michurina leaves. Vestnik of the Smolensk State Medical Academy. 2023;22(4):214–222. (In Russ.) DOI: 10.37903/vsgma.2023.4.29.

10. Pugacheva O. V., Panova K. E., Brezhneva T. A., Slivkin A. I. Selection of the optimal dosage form of chokeberry leaves Michurin. Biofarmatsevticheskii zhurnal. 2023;5:3–5. (In Russ.) DOI: 10.30906/2073-8099-2023-15-5-3-5.

11. Cvetanović A., Zengin G., Zeković Z., Švarc-Gajić J., Ražić S., Damjanović A., Mašković P., Mitić M. Comparative in vitro studies of the biological potential and chemical composition of stems, leaves and berries Aronia melanocarpa’s extracts obtained by subcritical water extraction. Food and Chemical Toxicology. 2018;121:458–466. DOI: 10.1016/j.fct.2018.09.045.

12. Suvorova I. N., Davydov V. V., Prozorovskiy V. N., Shvets V. N. Peculiarity of extract from Aronia melanocarpa leaves antioxidant action for the brain. Biomeditsinskaya Khimiya. 2005;51(1):66–71. (In Russ.)

13. Ipatova O. M., Prozorovskaya N. N., Prozorovsky V. N., Knzheva V. A., Baranova V. S., Gruzdeva A. E. Chokeberry leaf extract with biological activity and method for its preparation. Patent RUS No. 2171111. 27.07.2001. Available at: https://patentimages.storage.googleapis.com/74/9a/85/256ef6a82683f1/RU2171111C1.pdf. Accessed: 25.12.2024. (In Russ.)

14. Efenberger-Szmechtyk M., Nowak A., Czyżowska A., Śniadowska M., Otlewska A., Żyżelewicz D. ntibacterial mechanisms of Aronia melanocarpa (Michx.), Chaenomeles superba Lindl. and Cornus mas L. leaf extracts. Food Chemistry. 2021;350:129218. DOI: 10.1016/j.foodchem.2021.129218.

15. Ohgami K., Ilieva I., Shiratori K., Koyama Y., Jin X.-H., Yoshida K., Kase S., Kitaichi N., Suzuki Yu., Tanaka T., Ohno S. Anti-inflammatory effects of aronia extract on rat endotoxin-induced uveitis. Investigative Ophthalmology & Visual Science. 2005;46(1):275–281. DOI: 10.1167/iovs.04-0715.

16. Bell D. R. Gochenaur K. Direct vasoactive and vasoprotective properties of anthocyanin-rich extracts. Journal of Applied Physiology. 2006;100(4):1164–1170. DOI: 10.1152/japplphysiol.00626.2005.

17. Wu X., Gu L., Prior R. L., McKay S. Characterization of Anthocyanins and Proanthocyanidins in Some Cultivars of Ribes, Aronia, and Sambucus and Their Antioxidant Capacity. Journal of Agricultural and Food Chemistry. 2004;52(26):7846–7856. DOI: 10.1021/jf0486850.

18. Szopa A., Kokotkiewicz A., Kubica P., Banaszczak P., Wojtanowska-Krośniak A., Krośniak M., Marzec-Wróblewska U., Badura A., Zagrodzki P., Bucinski A., Luczkiewicz M., Ekiert H. Comparative analysis of different groups of phenolic compounds in fruit and leaf extracts of Aronia sp.: A. melanocarpa, A. arbutifolia, and A. prunifolia and their antioxidant activities. European Food Research and Technology. 2017;243:1645–1657.

19. State Pharmacopoeia of the Russian Federation. XIV edition. Moscow: Federalnaia elektronnaia meditsinskaia biblioteka; 2018. 7019 p. (In Russ.).

20. Shabunin S. V., Buzlama V. S., Ermakova T. I., Meshcheryakov N. P., Buzlama S. V., Buzlama A. V., Trutaev I. V., Dolgopolov V. N., Maksimova L. N. Screening of biostimulating and biocidal substances (adaptogens, bactericides and other drugs). Moscow – Voronezh: All-Russian Research Veterinary Institute of Pathology, Pharmacology and Therapy; 2006. 51 p. (In Russ.)

21. Trineeva O. V., Kolosova O. A., Slivkin A. I. Forecast of types of biological activity of the herb valerian doubty and volga with the help of the pass-online web resource. Proceedings of Voronezh State University. Series: Chemistry. Biology. Pharmacy. 2022;4:130–137. (In Russ.)

22. Kubasova E. D., Krylov I. A., Korelskaya G. V., Kubasov R. V. Plant tannins and some mechanisms of their pharmacological properties. Bulletin "Biomedicine & Sociology". 2022:7(4):5–11. (In Russ.) DOI: 10.26787/nydha-2618-8783-2022-7-4-5-11.

23. Azarova O. V., Galaktionova L. P. Flavonoids: anti-inflammatory mechanism of action. Khimiya rastitel’nogo syr’ya. 2012;(4):61–78. (In Russ.)