Application of TLC for the assessment of the flavonoid profile of sea buckthorn leaves during different phases of their preparation
Abstract
Sea buckthorn (Hippophae rhamnoides L.) is a perennial wind-pollinated dioecious berry shrub. The leaves, according to the literature, contain flavonoids, tannins, vitamins, organic, polyunsaturated fatty acids and amino acids and are a source of phenolic compounds (flavonoids, tannins, phenolic acids). According to the literature, sea buckthorn leaves contain various flavonoids, extracted mainly by alcohol-water mixtures. Thus, the composition of the flavonoid complex contains derivatives of quercetin, isorhamnetin and kaempferol (rutin, quercitrin, myricetin, luteolin, vitexin, narcissin, etc.). According to the authors, the annual supply of sea buckthorn leaves in the Greater Caucasus is 3-5 tons. Moreover, the leaves are an annually renewable by-product obtained during the procurement of the main raw material – fruits, which determines the relevance of comprehensive studies of the phytochemical composition of leaves within the framework of resource-saving technologies for the production of herbal medicines.
The purpose of our study was to substantiate the choice of conditions for the chromatographic separation of flavonoids from sea buckthorn leaves using TLC and investigation of their composition at different harvesting times.
Based on the parameters of chromatography efficiency, using a set of methods for visual assessment of chromatograms and their processing using the computer program “Densitometer Sorbfil”, the optimal conditions were selected for the separation of flavonoids from sea buckthorn leaves in a thin layer of sorbent: mobile phase ethyl acetate-glacial acetic acid-water (5:1: 1); developer – 5% alcohol solution of aluminium chloride when observed under UV light; 7 µl of extraction using HPTLC plates “Silica gel 60 F254” (10x15 cm). The presence of 18 biologically active substances with a flavonoid nature has been established. It has been established that when leaves are harvested from the beginning of June to the end of August (phases 1-3), the qualitative composition of pigments does not change. However, when the leaves are collected simultaneously with the fruits, in the TLC profile of the extracts, the zones of flavonoids were visually characterized by a smaller area and colour intensity than when they were harvested during the period of fruit formation. Now, research to substantiate the optimal phase of harvesting leaves from the point of view of their phytochemical composition and preserving the value of the fruit is ongoing. However, the method developed in the study can be recommended for inclusion in modern ND for sea buckthorn leaves of any phenological phase of plant life for the identification of a specific flavonoid profile of this raw material. In further studies, it is also necessary to evaluate the influence of the climatic conditions of plant growth, cultivation characteristics and varietal diversity on the reproducibility of the TLC profile of leaf flavonoids obtained in this study.
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References
Miheev A.M., Demenko V.I. Oblepiha. M.: Rosagropromizdat, 1990. 40 p. (In Russ.)
Hasenova A.B., Aralbaeva A.N., Utegalieva R.S., Mamataeva A.T., Mur-zahmetova M.K. Oblepiha krushinovidnaja (Hippophae rhamnoides L.) – istochnik bi-oaktivnyh veshhestv. Vestnik Almatinskogo tehnologicheskogo universiteta. 2020; 1: 82-86. (In Russ.)
Abdykalikova K.A., Nechiporenko L.P. Fitohimicheskij sostav nadzemnoj chasti oble-pihi krushinovidnoj. Estestven-nye nauki. 2008; 4: 104-107. (In Russ.)
Jaroszewska A., Biel W. Chemical composition and antioxidant activity of leaves of mycorrhized sea-buckthorn (Hip-pophae rhamnoides L.). Chilean Journal of Agricultural Research. 2017; 77: 155-162. http://dx.doi.org/10.4067/S0718-58392017000200155 (In Russ.)
Suryakumar G., Gupta A. Medicinal and therapeutic potential of Sea buckthorn (Hippophae rhamnoides L.). J Ethnophar-macol. 2011; 138(2): 268-278. https://doi.org/10.1016/j.jep.2011.09.024
Novruzov Je.N., Mamedov Z.G., Mustafaeva L.A., Mirjusifova H.M., Zejnalova A.M. Sostav i soderzhanie fla-vonoidov list'ev Hippophae rhamnoides L., proizrastajushhih v Azerbajdzhane. Himija rastitel'nogo syr'ja. 2018; 3: 209-214. https://doi.org/10.14258/jcprm.2018033772 (In Russ.)
Dharam P.A., Amrit K.S., Jyoti K., Tanveer N. Pharmacognostical Characteri-zation and Preliminary Phytochemical In-vestigation of Sea buckthorn (Hippophae rhamnoides L.) Leaves // Indo Global Jour-nal of Pharmaceutical Sciences. 2012. №2(2). рр. 108-113. https://doi.org/10.35652/IGJPS.2012.13.
Criste A., Urcan A.C., Bunea A., Furtuna F.R.P., Olah N.K., Madden R.H. et al. Phytochemical Composition and Bio-logical Activity of Berries and Leaves from Four Romanian Sea Buckthorn (Hippophae rhamnoides L.) Varieties. Molecules. 2020; 25(5): 1170. https://doi.org/10.3390/molecules25051170 (In Russ.)
Tkacz K., WojdyłoIgor A., Turkie-wicz P., Nowicka P. Triterpenoids, phenol-ic compounds, macro- and microelements in anatomical parts of sea buckthorn (Hip-pophaë rhamnoides L.) berries, branches and leaves. Journal of Food Composition and Analysis. 2021; 103: 104-107. https://doi.org/10.1016/j.jfca.2021.104107
Isachkin A.V., Zubik I.N., Potapova A.V., Ermakov M.A. Korreljacionnyj ana-liz fenofaz i fenointervalov u sortov oblepihi krushinovidnoj (Hippophae rham-noides L.) v kollekcii GBS RAN im. N.V. Cicina. Vestnik Kurskoj gosudarstvennoj sel'skoho-zjajstvennoj akademii. 2019; 2: 64-69. (In Russ.)
Korul'kin D.Ju., Abilov Zh.A., Muzychkina R.A., Tolstikov G.A. Prirod-nye flavo-noidy. Novosibirsk: Akad-emicheskoe izd-vo «Teo», 2007, 232 р. (In Russ.)
Lesovaja Zh.S., Pisarev D.I., Novikov O.O., Romanova T.A. Razrabotka metodiki kolichestvennogo opredelenija flavonoidov v trave manzhetki obyknovennoj Alchemilla vulgaris L.S.L. Nauchnye vedomosti Belgorodskogo gosu-darstvennogo universiteta. Se-rija: Medici-na. Farmacija. 2010; 22(93): 145-149. (In Russ.)
Drozdova I.L., Lupilina T.I. Razrabotka i validacija metodiki kolichestven-nogo opredelenija flavonoidov v trave ikotnika serogo. Kurskij nauchno-prakticheskij vestnik Chelovek i ego zdo-rov'e. 2016; 1: 106-112. (In Russ.)
Trineeva O.V., Rudaja M.A., Perova I.B., Slivkin A.I., Jeller K.I. Izuchenie profilja flavonoidov plodov oblepihi krushinovidnoj razlichnyh sortov. Biofarmacevticheskij zhurnal. 2021; 13(3): 22-29. (In Russ.)
Trineeva O.V., Rudaja M.A., Sliv-kin A.I. Issledovanie profilja flavonoidov plodov oblepihi krushinovidnoj razlichnyh sortov metodom tonkoslojnoj hromatografii. Sorbtsionnye i khromatograficheskie protsessy. 2020; 20(1): 79-86.
Trineeva O.V., Safonova I.I., Safonova E.F., Slivkin A.I. Opredelenie flav(In Russ.)onoidov i issledovanie vlija-nija uslovij hranenija na ih soderzhanie v plodah oblepihi metodom TSH. Sorbtsionnye i khromatograficheskie protsessy. 2012; 12(5): 806-813. (In Russ.)
Geiss F. Osnovy tonkosloinoi khro-matografii. M., Mir, 1999, 405 p. (In Russ.)
Rudakov O.B., Vostrov I.A., Fedorov S.V. i dr. Sputnik khromatografista. Metody zhidkostnoi khromatografii. Voro-nezh: Izd-vo «Vodolei», 2004, 528 p. (In Russ.)
Trineeva O.V., Slivkin A.I. Sravnitel'naya kharakteristika pigmentnogo sostava syr'ya i maslyanogo ekstrakta list'ev krapivy dvudomnoi. Razrabotka i registratsiya lekarstvennykh sredstv. 2016; 1(14): 142-148. (In Russ.)
Kovaleva N.A., Trineeva O.V., Chu-vikova I.V., Slivkin A.I. Development and validation of a procedure for quantitative determination of flavonoids in sea buck-thorn leaves by spectrophotometry. Bulle-tin of the Scientific Centre for Expert Eval-uation of Medicinal Products. Regulatory Research and Medicine Evaluation. 2023; 13(2): 216-226 https://doi.org/10.30895/1991-2919-2023-531 (In Russ.)
