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FATTY ACIDS COMPOSITION STUDY OF BIRTHWORT DUTCHMAN'S PIPE (ARISTOLOCHIA
CLEMATITIS L.) HERB AND ROOTS
Pohodina L.,
Postgraduate student of Department of Chemistry of Natural Compounds, National University of Pharmacy, Kharkiv, Ukraine Burda N.,
Doctor of Pharmaceutical Science, associate professor, associate professor of Department of Chemistry of
Natural Compounds, National University of Pharmacy, Kharkiv, Ukraine
Kyslychenko V.
Doctor of Pharmaceutical Science, professor, Head of Department of Chemistry of Natural Compounds,
National University of Pharmacy, Kharkiv, Ukraine
Abstract
The gas chromatography determined the fatty acids composition of birthwort Dutchman's pipe (Aristolochia clematitis L.) herb and roots. The results of experiment showed that palmitic acid prevails among the saturated fatty acids, whereas linoleic, linolic and oleic acids prevail among the non-saturated fatty acids. The bulk of the saturated fatty acids were contained in roots, and the non-saturated fatty acids were found mostly in herb.
Keywords: birthwort Dutchman's-pipe, fatty acids, gas chromatography.
Introduction
Aristolochia clematitis L. (birthwort Dutchman's pipe) is a perennial herbaceous plant relating to Aris-tolochiaceae family [1].
Birthwort Dutchman's pipe is met in Europe as well as in Asia, North America. The plant grows in wet places - along river and lake shores. Birthwort Dutchman's pipe is also found in flow meadows, brushlands and forests. It often appears as a weed [1, 2, 6, p. 199].
Birthwort Dutchman's pipe is not a compendia plant and is applied only in folk medicine of certain countries. The plant is popular in Chinese folk medicine and since ancient days serves for treatment of many diseases as antimicrobial, anti-inflammatory drug [1, 2]. Birthwort Dutchman's pipe herb is also used in cancer treatment [8, p. 594].
The chemical composition of birthwort Dutchman's pipe is presented by aristolochic acids, alkaloids, phenolic compounds, including flavonoids, coumarins, lignans, terpenic-type compounds [2, 3, p. 713-715, 5, p. 249-250, 6, p. 199-201].
One of the classes of bioactive substances possessing high pharmacological activity are the fatty acids. The fatty acids are known to produce anti-inflammatory effect [4, 7, p. 380-382]. Therefore, a study in fatty acids composition of birthwort Dutchman's pipe herb may prove to be expedient.
Moreover, the development of drugs on the basis of birthwort Dutchman's pipe herb with anti-inflammatory action may turn prospective, in particular, for cutaneous use.
Thus, a study of birthwort Dutchman's pipe herb and roots was feasible for their detailed phytochemical exploration.
Experimental
We studied the fatty acids composition of birthwort Dutchman's pipe herb and roots as collected before blossom and during the blossoming period in Kharkiv Region, Ukraine, in summer 2018.
The qualitative composition study and quantitative fatty acids content determination in herb and roots of birthwort Dutchman's-pipe were conducted on Selmichrom-1 gas chromatography unit with a flame ionization detector after preliminary methylation. Unit chromatographic column is made of stainless steel, 2.5 m long, internal diameter 4 mm. Stationary phase was Inerton after preliminary treatment with 10% diethy-leneglycol succinate (DEGS), carrier gas was nitrogen.
Chromatography unit operation parameters: temperature of column thermostat 180°C, of evaporator -230°C, of detector - 220°C. Nitrogen flow rate was 30 cm3/min. Sample volume of herb under study fatty acids methyl esters solution in hexane was 2 mm3.
The fatty acids methyl esters were obtained by modified Peisker method. Methylation was conducted with a mixture of chloroform, methanol and sulfuric
acid in relation 100:100:1. To 30-50 ^l of lipophilic fraction in glass ampoules 2.5 ml methylating mixture were added, ampoules were sealed and placed for 3 hours to thermostat at stable temperature of 105°C. Then the contents of ampoules was transferred to a test tube, a small amount of zinc sulfate powder and 2 ml each of purified water and hexane was added to extract methyl esters. The extract was stirred, settled and hexane phase was filtered off for chromatographic analysis.
In chromatograms the fatty acids were identified
by retention time as compared with standard samples of fatty acids. Reference samples were Sigma Ltd standards of saturated and unsaturated fatty acid methyl esters. Fatty acids content was calculated by normalization [9, p. 74-75, 10, p. 21-22].
Results and Discussion
The chromatograms of the fatty acid composition study in birthwort Dutchman's pipe herb and roots are specified in Figs. 1-4.
Fig. 1 The chromatogram of the fatty acid composition determination in birthwort Dutchman's pipe herb collected before blossom
Fig. 2 The chromatogram of the fatty acid composition determination in birthwort Dutchman's pipe herb collected in the period of blossoming
Fig. 3 The chromatogram of the fatty acid composition determination in birthwort Dutchman's pipe roots collected before blossom
Fig. 4 The chromatogram of the fatty acid composition determination in birthwort Dutchman's pipe roots collected in the period of blossoming
The results of the fatty acid composition study in birthwort Dutchman's pipe herb and roots are specified in Table 1.
Table 1
Birthwort Dutchman's pipe raw materials fatty acids composition
# Methyl ethers of fatty acids Samples
1 2 3 4
Content, %
1 С 12:0 lauric - - 8.00 3.30
2 С 14:0 myristic 0.83 0.74 0.22 0.10
3 С 14:1 myristoleic 0.16 0.08 0.65 0.80
4 С 16:0 palmitic 25.87 25.93 27.05 31.32
5 С 16:1 palmitoleic 0.65 0.53 0.07 0.07
6 Unidentified acid 0.25 0.17 0.10 0.04
7 С 18:0 stearic 4.32 4.78 1.68 1.77
8 С 18:1 oleic 9.52 10.70 10.85 9.98
9 С 18:2 linoleic 18.42 20.42 38.30 39.93
10 С 18:3 linolenic 36.98 34.35 11.44 11.27
11 С 22:0 behenic 1.40 0.87 0.71 0.72
12 С 24:0 lignoceric 1.60 1.43 0.93 0.70
Sum of unidentified acids 0.25 0.17 0.10 0.04
Sum of identified saturated fatty acids 34.02 33.75 38.59 37.91
Sum of identified non-saturated fatty acids 65.73 66.08 61.31 62.05
Note. 1 - birthwort Dutchman's pipe herb collected before blossom
2 - birthwort Dutchman's pipe herb collected in the period of blossoming
3 - birthwort Dutchman's pipe roots collected before blossom
4 - birthwort Dutchman's pipe roots collected in the period of blossoming
The retention time of fatty acids are specified in Table 2.
Table 2
_The retention time of fatty acids_
# Methyl ethers of fatty acids Sam] ales
1 2 3 4
Retention time, sec
1 С 12:0 lauric - - 100.47 100.38
2 С 14:0 myristic 160.93 160.59 156.80 157.48
3 С 14:1 myristoleic 204.81 203.14 201.52 201.00
4 С 16:0 palmitic 252.25 251.67 252.84 251.76
5 С 16:1 palmitoleic 314.17 314.02 311.05 310.17
6 Unidentified acid 336.42 336.57 342.03 342.36
7 С 18:0 stearic 409.04 410.29 411.23 408.47
8 С 18:1 oleic 485.13 484.36 485.52 485.83
9 С 18:2 linoleic 624.06 623.82 623.84 622.98
10 С 18:3 linolenic 845.27 844.45 843.24 842.64
11 С 22:0 behenic 1126.64 1125.51 1121.90 1127.13
12 С 24:0 lignoceric 1895.67 1893.64 1900.16 1896.01
Note. 1 - birthwort Dutchman's pipe herb collected before blossom
2 - birthwort Dutchman's pipe herb collected in the period of blossoming
3 - birthwort Dutchman's pipe roots collected before blossom
4 - birthwort Dutchman's pipe roots collected in the period of blossoming
As seen from the Table 1 data, in all objects of study 12 fatty acids were found, 11 of them being identified. In all birthwort Dutchman's pipe herb samples among the saturated acids palmitic acid prevailed, whereas most abundant non-saturated acids were lino-lenic, linoleic and oleic.
In comparison between birthwort Dutchman's pipe herb collected before blossom and in the period of blossoming we must point out that quantitatively the
saturated acids prevailed in herb collected before blossom (34.02 %), whereas the non-saturated acids were abundant in the blossoming herb (66.08 %).
The same trend was observed for roots as well.
The sum of the saturated fatty acids prevailed in birthwort Dutchman's pipe roots collected before and in the period of blossoming, the non-saturated acids -in both herb samples.
It must be noted that the content of palmitic and linoleic acids in root samples was higher than in birthwort Dutchman's pipe herb.
We also noted that the oleic acid content in herb collected before blossom was somewhat less than in roots collected within the same period (9.52 % and 10.85 % respectively). An inverse trend was observed for this acid in the blossoming herb and roots collected in the blossoming period (10.70 % and 9.98 % respectively).
The lauric acid was found only in birthwort Dutchman's pipe roots, its content being higher in roots collected before blossom (8.00 %).
Conclusion
Thus, the results obtained from the fatty acids composition study of birthwort Dutchman's pipe herb and roots collected at the different vegetation phases may be used and considered in the preparation of new substances and new drugs on the basis of these raw materials.
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DETERMINATION OF THE CONTENT OF ARBUTIN IN CRANBERRY FRUITS BY HPLC
Kuznietsova V.,
Candidate of Pharmaceutical Sciences, Associate Professor Dean of Pharmaceutical Department National University of Pharmacy Kharkiv, Ukraine Kyslychenko V. Doctor of Pharmacy, Professor Head of the Department of Chemistry of Natural Compounds
National University of Pharmacy Kharkiv, Ukraine
ОПРЕДЕЛЕНИЕ СОДЕРЖАНИЯ АРБУТИНА В ПЛОДАХ КЛЮКВЫ МЕЛКОПЛОДНОЙ
МЕТОДОМ ВЭЖХ
Кузнецова В.Ю.,
кандидат фармацевтических наук, доцент декан фармацевтического факультета Национальный фармацевтический университет
г. Харьков, Украина Кисличенко В.С. доктор фармацевтических наук, профессор заведующая кафедрой химии природных соединений Национальный фармацевтический университет
г. Харьков, Украина
