Determination of the optimal conditions for the extraction of anthraglycosides of alder buckthorn bark, development and validation of a method for their quantitative determination by high performance liquid chromatography

Objective: optimization of extraction conditions for anthraglycosides of alder buckthorn bark, development and validation of a method for their quantitative determination using high performance liquid chromatography. Materials and methods. The object of the study was alder buckthorn bark from different manufacturers. The work was performed on an Agilent 1100 liquid chromatograph complete with a G1311A four-solvent supply and degassing system, a G1315B diode array detector, a G1316A column thermostat, and a G1313A automatic sample injection device. Chromatographic analysis was carried out on a Zorbax SB-C18 reverse-phase column (4.6×250 mm, 5 μm) in a gradient elution mode of the mobile phase consisting of acetonitrile and highly purified water, adjusted to pH 2 with phosphoric acid. Results. As a result of the study, the optimal conditions for the chromatographic separation of anthraglycosides of alder buckthorn bark were determined. The optimal conditions for their extraction have been established: the grinding of plant raw materials is 250 µm, the extractant is ethyl alcohol 40%, the extraction temperature is 80℃, the extraction time is 90 minutes, the ratio of raw materials and extractant is 1:50. It has been proved that the developed method for the quantitative determination of anthraglycosides of alder buckthorn bark is specific, linear, correct, accurate and robust. The stability of the solution of the standard sample of glucofrangulin A and the obtained extract was established. Conclusion. A specific method for the quantitative determination of anthraglycosides of alder buckthorn bark has been developed, which can be used in quality control of this type of medicinal plant material.

1. Murav’yeva D.A., Samylina I.A., Yakovlev G.P. Pharmacognosy. Moscow: Medicine, 2002. 656 p. (in Russ.)

2. Alekseyeva G.M., Belodubrovskaya G.A., Blinova K.F., Goncharov M.Yu., Zhokhova E.V., Zelentsova A.B., MargnaU.V., Povydysh M.N., et al. Pharmacognosy. Medicinal raw materials of plant and animal origin. St. Petersburg: SpetsLit, 2010. 863 p. (in Russ.)

3. Roudbaraki, S. J., Nori-Shargh D. Analysis of the volatile constituents of Frangula alnus Mill. from Iran.Russian Chemical Bulletin. 2016;65(11): 2770-2772. DOI: 10.1007/s11172-016-1652-0.

4. Arsenijevic J., Drobac M., Slavkovska V., Kovacevic N. Anatomical analysis and phytochemical screening of Frangula rupestris (Scop.) Schur (Rhamnaceae). Botanica Serbica. 2018;42(2):231-239. DOI: 10.5281/zenodo.1468339.

5. Romanyuk A.A., Moiseyev D.V. Alder buckthorn bark: component composition, pharmacological properties, standardization. Vestnik farmatsii. 2020;2(88):95-104 (in Russ.). EDN: XWYMHG.

6. Bacha A., Jemel I., Moubayed N., Abdelmalek I. Purification and characterization of a newly serine protease inhibitor from Rhamnus frangula with potential for use as therapeutic drug. Journal of Biotech Research. 2017;7(2):1-13. DOI: 10.1007/s13205-017-0764-z.

7. Izhaki I. Emodin-a secondary metabolite with multiple ecological function in higher plants. New Phytologist. 2002;155(2):205-217. DOI:10.1046/j.1469-8137.2002.00459.x.

8. Sydiskis R., Owen D., Lohr J., Rosler K., Blomster R. Inactivation of enveloped viruses by anthraquinones extracted from plants. Antimicrobical Agents Chemotherapy. 1991;35(12):2463-2466. DOI: 10.1128/AAC.35.12.2463.

9. Manojlovic N., Solujic S., Sukdolak S., Milosev M. Antifungal activity of Rubia tinctorum, Rhamnus frangula and Caloplaca cerina. Fitoterapia. 2005;76(2):244-246. DOI: 10.1016/j.fitote.2004.12.002.

10. Abad M., Ansuategi M., Bermejo P. Active antifungal substances from natural sources. Archive for Organic Chemistry. 2007;7:116-145. DOI: 10.3998/ark.5550190.0008.711.

11. Stef D., Gergen I., Trasca T., Harmanescu M., Lavinia S., Ramona B., Heghedus M. Total antioxidant and radical scavenging capacities for different medicinal herbs. Romanian Biotechnological Letters. 2009;14(5):4704-4707. URL: http://www.rombio.eu/rbl5vol14/12.pdf.

12. Marchenko S.I. State Pharmacopoeia of the Republic of Belarus. Volume №2. Quality control of substances for pharmaceutical use and medicinal plant raw materials. Molodechno: Victory, 2016. 1368 p. (in Russ.)

13. Emshinova S.V., Potanina O.G., Budanova E.V., Chistyakov V.V. State Pharmacopoeia of the Russian Federation. VolumeIV. Moscow: Medicine, 2018. 7019 p. (in Russ.)

14. Tulegenova A.U. State Pharmacopoeia of the Republic of Kazakhstan. Volume No 3. Astana: Zhibekzholy, 2014. 872 p. (in Russ.)

15. Marianne E.K., editor. Eurорeаn Рhаrmаcороeiа:EDQM. 10th Editiоn. URL: https://pheur.edqm.eu/home

16. Romanyuk A.A., Moiseyev D.V. Optimization of conditions for extraction of frangulin A into adler buckthorn brittle by high performance liquid chromatography. Vestnik Bashkirskogo gosudarstvennogo meditsinskogo universiteta. 2019;(4):291-295 (in Russ.). EDN: QUXDIR.

17. Sheryakov A.A. State Pharmacopoeia of the Republic of Belarus. Volume №1. General methods of drug quality control. Molodechno: Victory, 2012.1220 p. (in Russ.)

18. Samylina I.A., Moiseyev D.V., Marchenko S.I., Veremchuk O.A., Moiseyeva A.M., Sorokina A.A. Harmonization of methodological approaches to the standardization of pharmacopoeial types of plant materials containing flavonoids. Pharmacy. 2020;5(69):5-11 (in Russ.). DOI: 10.29296/25419218-2020-05-01. EDN: PLLZEG.

19. Epshteyn N.A., Sevast’yanova V.L., Koroleva A.I. Robustness study in the validation of HPLC and UPLC methods: a modern approach including risk analysis. Drug development and registration. 2018;4:96-109 (in Russ.). EDN: VYLMKT.