CC BY
55
УДК: 615.322:581.45:582.46:54.061/062
IU. V. IUDINA
National university of pharmacy, Kharkov, Ukraine
STYDING OF NATURE AND QUANTITY OF POLYPHENOLIC SUBSTANCES IN GINKGO BILOBA LEAVES
Resume: This article presents the results of the analysis of the qualitative and quantitative content of polyphenolic compounds in the leaves of Ginkgo biloba gathered during the vegetation period on the territory of the Botanical garden KNU named. Karazin. In the leaves were found rutin, quercetin, cafe and quinic acid, and defined their quantitative content. Deadline was set in raw materials, when the content of these substances reaches maximum values. Keywords: Ginkgo biloba, quinic acid, analysis, substances, Botanical garden KNU named. Karazin.
Chemical composition of Ginkgo biloba leaves begins to investigate from the 80-th of last century. However in a plant always take place biochemical processes, thus its chemical compose constantly changes. Moreover chemical compose of raw material depends on climatic features and cultivating technology. So at development of pharmaceuticals based on herbal raw materials is very important to determine the harvesting period when the content of BAS achieves its maximum. Antioxidant properties of Ginkgo biloba preparations cause polyphenolic substances. Thus it was conducted a row of experiments in order to determine the period of vegetation of Ginkgo biloba when the content of BAS gets its maximum in Ukrainian climatic conditions.
Materials and methods of research. As object of research was choose Ginkgo biloba leaves harvested during vegetation period in Botanical garden of Karazin's Kharkov National University.
For determination of polyphenolic compounds in Ginkgo biliba leaves was used next technique. About 1g of ginkgobiloba leaves grinded up to 1-2 mm was poured by 70% alcoholatratio of raw material to extragent 1:10 and leave for 1 hour for extraction. Than flask with the extract was united to a reverse refrigerator and heated to a
light boiling during 2 hours. After cooling extract was filtrated. Content of polyphenoloic substances were determinate by well-knownmethods [2,3,4]. Qualitive content of flavonoids. Analysis of the qualitative content of flavonoids was performed by one-dimensional ascending paper chromatography chromatographic grade "Filtrak" № 7, 11 in a solvent system 15% acetic acid, butanol: acetic acid: water 4:1:5 (4:1:5 BAW) and the plates "Silufol UV-254", "Silufol UV-366" in the solvent system BAW 4:1:5.
By the analysis of qualitative content of polyphenolic compounds in Ginkgo leaves were selected method for their quantitative determination [3,4].
Results and discussions
Research ofqualitative content of polyphenolic compounds in Ginkgo leaves. To determine the nature of the polyphenolic compounds was carried out colored sedimentary reactionson these compounds, the results are given in Table 1
As can be seen from Table 1, the presence of polyphenolic compounds observed during the summer-autumn period, as evidenced by the intense black and green color of 70% Ginkgo biloba leaves alcohol extract at the reaction with iron III chloride.
Table 1
№ Reaction Growing season
beginning of the growing season Efflorescence formation of fruit scattering of fruits
1 cyanidinic test red color of the solution, the organic phase is less intensely colored red color of the solution, the organic phase is less intensely colored red color of the solution, the organic phase is less intensely colored red color of the solution, the organic phase is less intensely colored
2 1% alcohol solutionof КОН yellow color of the solution transferintoorange yellow color of the solution transferintoorange yellow color of the solution transferintoorange yellow color of the solution transferintoorange
3 solution of acetic lead yellow precipitate yellow precipitate yellow precipitate yellow precipitate
4 solution of ferric chloride (Ill) black - green color of the solution black - green color of the solution black - green color of the solution black - green color of the solution
Qualitative analysis of polyphenolic compounds content in the leaves of Ginkgo biloba during the growing season By cyanidinic test was found that flavonoid compounds in Ginkgo leaves are more in the form of aglycones than glycosides, as evidenced by a less intense color of the
organic phase. By the intensity of color was concluded that their major content observed in September-October To clarify the nature of polyphenolic compounds were performed chromatographic studies of 70% Ginkgo biloba leaves ethanol extract during the growing season. Chromatogramesare shown in Figure 1.
Rf 1
0,9 0,8 0,7 0,6 0,5 0,4 0,3, 0,2 0,1
0 0 0
8Ö 0 C3 G
7 O o o o
6 0 0 0 0
5 ♦ 0 0 0
4^ CJ C
C33 fl CD A CD n
\J 0 \J O 0
°2 °3 °4
□
b.
Rf
1
0,9
7 0
0,8 0 0 0
6 O o o O o
0,7
0,6 5 ^ /D
4 G u o CJ>
0,5 o Q o
0,4 £> CP CO e>
0,3,
0,2 2 0 CD o
0,1 1
■Q o .
1 2 3 4 5
o
Q
'0
Fig 1 a
Fiq 1 b
Figure 1 - Chromatogrames of Ginkgo biloba leaves extracts a - BAW 4:1:5 b -15% acetic acid 1- beginning of the growing season, 2- efflorescence, 3-formation of fruit, 4- scattering of fruits Test samples: 5 - chlorogenic acid, 6 - caffeic acid, 7 - quercetin, 8 - rutin
In solvent system 15% acetic acid was detected compounds 7, BAW 4:1:5 - 9 compounds. Chromatogram were tested in daylight (polyphenolic compounds shown as yellow-brown spots in all solvent systems) and in UV-light at ammonia vapors development spots change their color depending on the nature of the substance (table 2).
Table 2 - The results of chromatographic analysis of polyphenolic compounds isolated from Ginkgo leaves
8
1
a
№ Solvent system
15% acetic acid BAW 4:1:5
Rf Color of stains Rf Color of stains
Daylight UV-light. with NH3 vapors Daylight UV-light. with NH3 vapors
1. 0,02 yell. yell. yell. 0,05 lightyell. blue violet
2. 0,14 yell. yell. yell. 0,21 yell. yell. yell.
3. 0,43 lightyell. dark yell. dark yell. 0,32 lightyell. yell. yell.
4. 0,50 yell. blue blue 0,53 lightyell. yell. dark yell.
5. 0,59 dark yell.. brown brown 0,59 dark yell. brown brown
6. 0,77 yell. blue blue 0,65 lightyell. dark yell. dark yell.
7. 0,84 dark yell. blue blue 0,74 yell. blue blue
8. 0,82 yell. yell. yell.
9. 0,94 yell. blue blue
Comparing the samples of extract strains with test substances and the values of Rf were identified flavonoids rutin, quercetin, kaempferol, mirytcetyn and phenolcarboxylic acids: caffeic and chloragenic. By the intensity of strains color concluded that rutin and quercetin are dominant substances in the extract.
Quantitative analysis of the polyphenol compounds content in Ginkgo biloba leaves. Was conducted spectrophotometry study for determination of quantitative content of polyphenolic compounds in the Ginkgo leaves. UV-spectra of Ginkgo leaf extract and quercetin standard are shown in Figure 2.
D 1
0,9 -
0,8 -
0,7 -
0,6 -
0,5 -
0,4 -
0,3 -
0,2 -
0,1 -
0 -
260 270 280 290 302 310318,8330341,5350364,7375 378 380 390 402L,nm
Figure 2 - UV absorption spectra of standard sample of quercetin (1) and investigated Ginkgo biloba leaf extract (2)
The spectrum of a standard quercetin sample (1) has a maximum absorption at a wavelength of 375±1 and 303±1 nm range tested extracts of Ginkgo leaves (2) has a maximum of absorption at a wavelength of 302±1 nm and 374±1 nm. Calculation of polyphenolic compounds amount in Ginkgo leaves during the growing season are given in Table 3.
Table 3
Vegetation period The quantitative content of polyphenolic compounds in the Ginkgo leaves, % (n=5)
beginning of the growing season 1,7±0,05 Еотн 2 5%
efflorescence 2,4±0,03 Еотн -, -2%%
formation of fruit 3,0±0,04 Еотн= 1,84%%
scattering of fruits 3,4±0,08 Еотн= 2,67%%
subsidence of leaves 2,1±0,04 Еотн= 2,15%%
Amounts of polyphenolic compounds in Ginkgo biloba leaves during the growing season
Research has shown that polyphenolic compounds gradually
accumulated in leaves during the growing season and their maximum observed in September and October, and is 3,4±0,08% in terms of quercetin. Then in November, the content thereof is reduced and takes 2,1 ± 0,04%. Thus the results of the research it can be concluded that September-October is the optimal period of harvestingofrawmaterial, when the amount of polyphenolic compounds reaches maximum values.
REFERENCES
1 Герiатричний лтарський 3aci6 «Пногран» //Ю.В. Щирова, В.Г Дем'яненко, Л.В. Деримедвщь, С.М. Дроговоз. - Пат. Украши 58880 А, МК1 А61К35/78, № 2002119103; заявл. 15.11.02; опубл. 15.08.03. Бюл. №8, 2003 р.
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Conclusions
1. Researches of Ginkgo biloba leaves polyphenolic compounds nature have shown their presence throughout the growing season. Determined that Ginkgo biloba leaves contains flavonoids quercetin, rutin, which are dominant, and kaempferol, mirytsetyn, caffeic and chlorogenic acids.
2. Spectrophotometric study of quantitative content of polyphenolic compounds in Ginkgo leaves shown that the term of raw materials harvesting when the content of polyphenolic compounds reaches maximum value (3,4 ± 0,08%), falls in September-October.
7 Gu Hiaomi, Li Lei Measurment of rutin in leaves of Ginkgo biloba with revers-phase HPLS // Sepu. - 1995. - Vol. 13, №3. -P.216-217.
8 Isolation of amentoflavone from Ginkgo biloba / A. Lobstein-Guth, F. Brianacon-Scheid, C. Victoire, M. Haag- Berrurier, R. Anton // Planta Med. - 2006. - Vol. 54. - №6. - P. 555-556.
9 Kaempferol coumarol glucorhamnoside from Ginkgo biloba/Anton Robert, Nars Chamel, Haag-Berrurier Michelline, Lobstein-Guth Annelise // Phutochemistry. - 2008. - Vol.25. - №3. - P.770-771.
10 You Soug, Yao Xinsheng, Shluyang Yaoxueyau Flavon glucosides from the leaves of Ginkgo biloba // Xuebao. - 2008. - Vol.6. №4. - P. 284-285.
Ю.В. ЮДИНА
Харьков цаласыныц Украина мемлекетт'к фармацевтикалы;университет'1
ГИНКГО БИЛОБА ЖАПРА^ТАРЫНДАгЫ ПОЛИФЕНОЛЬДЫ кОСЫЛЫСТАРДЫн САПАЛЫк ЖэНЕ
К0ЛЕМД1К к¥РАМЫН аныктау
Туйш: Бул ма^алада, Карзин атындагы ХМУ-дыц ботаникалыщ багында барлыщ вегетациялыщ кезецЫде жиналган гинкго билоба жапра^тарыныц ^урамындагы полифенольды ^осылыстардыц сапалыщ жэне сандыщ сараптамасы келтiрiлген. Жапыра^тарда рутин, кверцетин, кофелт жэне хлорагендiк ^ыш^ылдары, сонымен ^атар олардыц сандыщ 1^урамы аны^талды. Бул заттардыщ ^урамыныц ец жогаргы кврсеткiшiтiк кезецЫщ, ши^заттыщ дайындалатын мерзiм аны^талды Тyйiндi сездер: гинкго билоба, кверцетин, сараптама, Карзин атындагы ХМУ-дыц ботаникалыщ ба^.
Ю.В ЮДИНА
Национальный фармацевтический университет, Харьков, Украина
ИЗУЧЕНИЕ КАЧЕСТВННОГО И КОЛИЧЕСТВЕННОГО СОДЕРЖАНИЯ ПОЛИФЕНОЛЬНЫХ СОЕДИНЕНИЙ
В ЛИСТЬЯХ ГИНКГО БИЛОБА
Резюме:В данной статье приведены результаты анализа качественного и количественного содержания полифенольных соединений в листьях гинко билоба, собранных на протяжении периода вегетации на территории ботанического сада ХНУ им. Каразина. В листьях были обнаружены рутин, кверцетин, кофейная и хлорагеновая кислоты, а так же определено их количественное содержание. Был установлен срок заготовки сырья, когда содержание этих веществ достигает максимальных значений.
Ключевые слова: гинко билоба, ботанический сад ХНУ им. Каразина, кверцетин, исследование, вещество
