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28PHARMACEUTICS
THE STUDY OF THE MINERAL COMPOSITION OF HELIOPSIS HELIANTHOIDES
Pavlenko-Badnaoui M.,
PhD student of the Department of Chemistry of Natural Compounds of the National University of Pharmacy, Ukraine
Protska V.,
PhD, assistant of the Department of Chemistry of Natural Compounds of National University of Pharmacy, Ukraine
Zhuravel I.
Doctor of Pharmaceutical Science, professor, professor of Department of Chemistry of Natural Compounds of National University of Pharmacy, Ukraine
Abstract
With the help of the atomic absorption spectroscopy method, 19 macro- and microelements were identified in Heliopsis helianthoides roots, leaves, flowers and seeds.
The greatest amount of mineral elements was accumulated in the leaves of Heliopsis helianthoides (8630.33±215.76 ^g/100 g). In other types of raw materials studied, the total content of these compounds was 2 -3 times lower.
Quantitatively, potassium, magnesium, and silicon prevailed in all types of raw materials, the highest content of which was observed in the Heliopsis helianthoides leaves - 4450.00±111.25 ^g/100 g, 1780.00 ± 44.50 ^g/100 g and 1240.00±31.00 ^g/100 g, respectively.
Iron (120.00±3.00 ^g/100 g) accumulated in significant amounts among the essential elements of Heliopsis helianthoides in the roots, and zinc (35.00 ± 0.88 ^g/100 g) and strontium (5.30±0.13 ^g/100 g) in the leaves.
The content of heavy metals was within the limit of maximum permissible concentrations for medicinal plant materials and food products.
Keywords: atomic absorption spectroscopy, micro- and macroelements composition, Heliopsis helian-thoides, Astersceae.
Introduction
The genus Heliopsis L. belongs to the Asteraceae L. family. Mexico and Latin America are considered to be the habitat of plants of this genus [1].
Heliopsis helianthoides (L) Sweet. is a perennial plant. The root system of this plant is fibrous, although in arid areas of growth it can be modified into a root crop. Its stem is erect, pubescent. Leaves are petiolate, opposite, oval or ovate, with a toothed margin, pubescent. Large flowers of bright yellow inflorescences form a basket. The fruit is a small flat achene of gray-brown colour [1].
Indigenous peoples from Mexico and South America used Heliopsis helianthoides to relieve toothache and muscle pain, and to treat infectious diseases, as an anti-ulcer and insecticide remedy [2-5].
A group of scientists from Cuba, Chile and Mexico found that ethanolic extracts of Heliopsis helian-thoides roots in experiments on rats showed analgesic, antinociceptive, anxiolytic, diuretic activities and suppressed the production of inflammatory process mediators prostaglandins and leukotrienes, inhibited cycloox-ygenase and 5-lipoxygenase enzymes [2, 4, 6]. Mexican scientists in studies on rats found that ethanol and dichloromethane extracts of the Heliopsis roots exhibited a vasodilating effect [6].
Mineral elements play significant role in ensuring the normal functioning of the human body. It is well known that potassium and sodium support the body's homeostatic balance and affect blood pressure. Calcium is involved in the transmission of nerve impulses,
is a structural element of connective tissue. Iron in hemoglobin and cytochrome provides cellular respiration. Magnesium, copper and zinc are cofactors of enzymes and hormones, a number of biochemical processes occur in the body with their participation [7, 8].
At the same time, when assessing the quality of medicinal plant materials, the content of heavy metals, in particular, lead, cobalt, mercury, etc. is equally important. These elements tend to accumulate in the body tissues, and their excess leads to serious consequences. In chronic poisoning with mercury compounds, mental disorders occur, as well as encephalopathy, visual and hearing impairment, tremor, tachycardia, nephrosis, and gastroenteritis are manifested [9, 10]. Excess cadmium in the body leads to osteoporosis, anemia, hypertension, nephro- and cardiopathy, and liver damage. Toxic doses of lead interfere with hematopoietic processes, cause anemia, nephro- and encephalopathy [9, 10]. Intoxication with arsenic compounds manifests damage to the nervous system, hemolysis of erythrocytes, the development of severe forms of cardiac, renal and hepatic failure [9, 10]. Therefore, the content of heavy metals in medicinal plant raw materials and food products is regulated by the Law of Ukraine "On Basic Principles and Requirements for Safety and Quality of Food Products" and the Order of the Ministry of Health of Ukraine "On Approval of State Sanitary Regulations and Rules" Medical Requirements for the Quality and Safety of Food and Alimentary Raw Materials" No. 1140 dated January 09, 2013 [11, 12].
The aim of our work was the study of the content of mineral elements, heavy metals in particular, in the plant raw material of Heliopsis helianthoides.
Material and methods
The shredded, air-dried roots, leaves, flowers, and seeds of Heliopsis helianthoides, collected in 20172018 on the territory of Kharkiv region (Ukraine) were used for this study.
The study of the qualitative composition and quantitative content of mineral elements was carried out on the basis of the State Scientific Institution "Institute for Single Crystals" of the National Academy of Sciences of Ukraine (Kharkov). The element composition of the raw material was analyzed by atomic absorption spec-troscopy with atomization in an air-acetylene flame on the CAS-120 device [13, 14].
Samples of plants were charred in a muffle furnace at a temperature of about 500°C after pretreatment with dilute sulfuric acid. Evaporation of the samples was carried out from craters of graphite electrodes in a discharge of an AC arc with a force of 16 A at an exposure of 60 s. An IVS-28 device was used to excite the source of the spectrum. They were recorded on a photographic film with a spectrograph, with a diffraction grating of 600 lines/mm and a triple-lens slit illumination system [13, 14].
Calibration graphs were constructed by using standard solutions of metal salts (ISORM - 23 - 27) within the limits of the corresponding measured concentrations of elements. The dissolution of the elements was carried out with the use of chemically pure qualifying chemicals and twice purified water [13, 14].
Measurements were carried out using MF-4 microphotometer in the absorption spectrum from 240 nm to 347 nm compared with standard samples of a mixture of elements [13, 14].
Results and discussion
According to the results of the analysis, 19 mineral elements were identified and their quantitative content was determined in Heliopsis helianthoides roots, leaves, flowers and seeds.
The maximum content of macro- and microelements was observed in the Heliopsis helianthoides leaves - 8630.33±215.76 ^g/100 g. At the same time, they accumulated almost in a 2 times less quantity in the roots (3937.38±98.43 ^g/100g), in flowers (2961.15±74.03 ^g/100 g) and seeds (2621.61±65.54 ^g/100 g) - three times less compared to their content in the leaves of this plant. The results of analyzes are presented in the table 1.
Potassium quantitatively dominated among certain elements, which was characteristic of all the objects studied. The content of this mineral in the Heliopsis helianthoides roots was about 47% of the total content of all identified elements and comprised 1860.00±46.50 ^g/100 g. The share of potassium in the leaves and the seeds comprised about 52% (4450.00±111.25 ^g/100 g) and 60% (1560.00±39.00 ^g/100 g), respectively. At the same time, the content of this element in the leaves was 2-4 times higher than its content in other parts of the studied plant. In the Heliopsis helianthoides flowers the potassium content was about one third of the total mineral content.
Tabl. 1
Qualitative composition and quantitative content of macro-to-micronutrients in the raw material of
№ chemical element Content of mineral elements, ^g / 100 g
Roots Leaves Flowers Seeds
1 Ferrum 120.00±3.00 90.00±2.25 14.00±0.35 7.20±0.18
2 Silicium 730.00±18.25 1240.00±31.00 370.00±9.25 180.00±4.50
3 Phosphorus 130.00±3.25 270.00±6.75 280.00±7.00 210.00±5.25
4 Aluminum 160.00±4.00 80.00±2.00 32.00±0.80 12.00±0.30
5 Manganese 4.80±0.12 8.90±0.22 10.20±0.26 3.00±0.08
6 Magnesium 200.00±5.00 530.00±13.25 280.00±7.00 170.00±4.25
7 Nichel 0.30±0.01 0.09±0.01 0.09±0.01 0.09±0.01
8 Molybdenum 0.08±0.01 0.14±0.01 0.13±0.01 0.12±0.01
9 Calcium 650.00±16.25 1780.00±44.50 930.00±23.25 420.00±10.50
10 Cuprum 1.00±0.03 0.90±0.02 0.83±0.02 1.00±0.03
11 Zinc 0.60±0.02 35.00±0.88 16.70±0.42 9.00±0.23
12 Sodium 80.00±2.00 140.00±3.50 93.00±2.33 48.00±1.20
13 Potassium 1860.00±46.50 4450.00±111.25 930.00±23.25 1560.00±39.00
14 Strontium 0.60±0.01 5.30±0.13 4.20±0.11 1.20±0.03
Total mineral content 3937.38±98.43 8630.33±215.76 2961.15±74.03 2621.61±65.54
Content of heavy metals
15 Plumbum <0.03 <0.03 <0.03 <0.03
16 Cobalt <0.03 <0.03 <0.03 <0.03
17 Cadmium <0.01 <0.01 <0.01 <0.01
18 Arsenium <0.01 <0.01 <0.01 <0.01
19 Mercury <0.01 <0.01 <0.01 <0.01
The calcium content in the raw material of corresponding object. The high content of this element Heliopsis helianthoides was from 16% to 31% of the was observed in the leaves of the studied plant -total content of all mineral elements in the 1780.00±44.50 ^g/100 g, which was 2.5 times less than
the potassium content in this type of the raw material. The content of potassium and calcium in the Heliopsis helianthoides flowers was almost at the same level and amounted to 930.00±23.25 ^g/100 g. At the same time, in the flowers calcium was almost half as much as in the leaves. The content of this mineral in the roots and seeds of Heliopsis helianthoides was 650.00±16.25 ^g/100 g and 420.00±10.50 ^g/100 g, respectively. The high content of silicium was recorded in the Heliopsis helianthoides leaves - 1240.00±31.00 ^g/100 g, which amounted to about 14% of the total mineral content in this plant material type. In the Heliopsis helianthoides roots this element was accumulated 1.7 times less, in flowers - more than 3 times less, in seeds - almost 7 times less. The numerical value of the silicium content in the Heliopsis helianthoides roots was 730.00±18.25 ^g/100 g, in flowers and seeds - 370.00±9.25 ^g/100 g and 180.00±4.50 ^g/100 g, respectively.
The largest amount of magnesium was accumulated in the Heliopsis helianthoides leaves and amounted to 530.00±13.25 ^g/100 g. The content of this element in the flowers of the studied plant was almost half as high - 280.00±7.00 ^g/100 g. The Heliopsis helianthoides roots and seeds accumulated almost equal amount of magnesium - 200.00±5.00 ^g/100 g and 170.00±4.25 ^g/100 g, respectively.
A high iron content is noted in the Heliopsis helianthoides roots (120.00±3.00 ^g/100 g). Almost the same maximum amount of phosphorus accumulated in the leaves and flowers of this plant and amounted to 270.00±6.75 ^g/100 g and 280.00±7.00 ^g/100 g. In addition, the leaves also contained a significant quantity of sodium (140.00±3.50 ^g/100 g) and zinc (35.00±0.88 ^g/100 g). It was noted that the zinc content in all the parts of this plant did not exceed 1 ^g/100 g in the roots and seeds contained 1.00±0.03 ^g/100 g of cuprum each, in the leaves - 0.90±0.02 ^g/100 g, in flowers - 0.83±0.02 ^g/100 g. Strontium quantitatively prevailed in the leaves (5.30±0.13 ^g/100 g) and flowers (4.20±0.11 ^g/100 g) of Heliopsis helianthoides
The content of heavy metals in all analyzed types of raw materials did not exceed the maximum permissible concentrations in accordance with the requirements of current regulatory documents.
Conclusion
19 mineral elements were identified and their content determined using the method of atomic absorption spectrometry in the roots, leaves, flowers, and seeds of Heliopsis helianthoides. It was established that the largest number of mineral elements was accumulated in the leaves of Heliopsis helianthoides and amounted to 8630.33±215.76 ^g/100 g.
Among the studied elements, potassium, calcium, and silicium prevailed in all types of raw materials, the maximum content of which was recorded in the Heliopsis helianthoides leaves. The content of potassium in this type of raw material was 4450.00±111.25 ^g/100 g, the content of magnesium and silicium -530.00±13.25 ^g/100 g and 1240.00±31.00 ^g/100 g, respectively.
A significant amount of iron accumulated in the roots of Heliopsis helianthoides (120.00±3.00 ^g/100
g), in the leaves and flowers - phosphorus (270.00±6.75 ^g/100 g and 280.00±7.00 ^g/100 g, respectively).
The obtained results can be used in the standardization of Heliopsis helianthoides raw materials and in the development of medicinal herbal drugs based on it.
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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
