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https://doi.org/10.29013/AJT-21-1.2-45-48
Kutlimurotova R. Kh., Pulatova L. T., Khaitbaev A. Kh., National University of Uzbekistan E-mail: nigora.qutlimurotova@mail.ru
QUANTITATIVE ANALYSIS OF MICRO AND MACROELEMENTS IN LEAVES OF THE PLANT ASARUM EUROPAEUM L. BY METHOD OF MASS SPECTROMETRY
Abstract. The quantitative analysis of micro-and macroelements in the leaves of the plant Asarum europaeum L was carried out by inductively coupled plasma mass spectrometry (ICP-MS). The sum of flavonoids was investigated by spectrophotometry method. It was determined: that the most distributed macroelements are potassium, calcium and magnesium, while the most wide spread trace elements are manganese, barium and zinc. At the same time it was determined that on the intensity of elements accumulation in plant the place of in's growth has influenced.
Keywords: mass spectrometric analysis, European hoof, micro and macroelements, extraction.
At present time population has shown great at- anti-inflammatory and antispasmodic effects. Infu-
tention to medicines and biologically active substances obtained from natural plant raw materials. Despite the long period of using in folk and official medicine of domestic medicinal plants, their true potential is far from fully revealation. One of these medicinal plants is Asarum europaeum L.
Asarum europaeum L is a perennial herbaceous plant from the family Kirkazonaceae with a rather brownish, thick, recumbent stem (up to 30 cm long) covered with short pressed hairs and a creeping rhizome. The plant has a peculiar smell. The leaves are simple, without stipules, long-stemmed, whole-edged, kidney-shaped (5-8 cm in diameter) with well-defined veins. The flowers are fleshy, small (about 1 cm), bisexual, solitary, axillary, on short drooping pedicels, regular (actinomorphic), red-brown, usually formed at the ends of young spring shoots. The perianth is 3-mem-bered, and has a peculiar lateral shape. It blooms in April and May and is found in botanical gardens [1-5].
Asarum europaeum is used in folk medicine as an emetic, expectorant, possezing with antibacterial,
sion of leaves is drunk at diseases from the stomach, liver, kidneys; decoction - at heart diseases, alcoholism and poisoning by poisonous mushrooms, nervous excitement, migraines, as a diuretic and dropsy, jaundice, malaria, eczema, epilepsy. Besides, a decoction of the roots with rhizomes of wild ginger is used as a diuretic for kidney disease. Especially effective is the decoction of underground organs on milk [6]. There are evidences that decoction of rhizomes has increased the amount of male seed. To enhance of sexual desire and potency, an infusion of rhizomes on grape syrup is used. The water infusion of the plant's raw materials is drunked at diseases of the stomach, liver, and kidneys; the decoction is used at heart diseases, alcoholism and poisoning with poisonous mushrooms (as an emetic), nervous excitement, migraines, as a diuretic for dropsy, as well as for jaundice, malaria, eczema, and epilepsy [6].
In Uzbekistan this plant is called ungulate, adapted to the conditions of Uzbekistan, and is grown in the Tashkent Botanical Garden. In this study, the
chemical composition of the leaves of this plant was investigated.
The experimental part
Used reagents: multi-element standard on 29 elements for MS standard-Hg (mercury), nitric acid (h / h), hydrogen peroxide (h / h), deionized water, argon (purity 99.995%).
As an object of research the leaves of European ungulate harvested after the end of fruiting in Uzbekistan (end of June 2020) were used. The raw material was dried to an air-dry state, was packed in double paper bags and has stored in a cool, dry place.
40% ethyl alcohol was used as extractant for extract ion the total sum of flavonoids from raw materials, which according to previous studies, along with 70% ethyl alcohol provides the greatest completeness of flavonoid extraction from the leaves of Asarum europaeum L. [3].
Used instruments and containers: ICP MS NAVISI0N-2000, mass spectrometer, microwave
disintegrator (Germany) or similar autoclave Teflon measuring flasks.
Method of quantitative determination of the amount of flavonoids in the extract
1 ml (exact volume) of the extract is placed in measuring flask with capacity 25 ml, 3 ml of 2% aluminum chloride solution, one drop of diluted acetic acid, were added and the volume of the solution was by 95% ethanol to the mark. After 40 minutes the solution is filtered through paper filter. The optical density ofthe resulting solution is measured on a spectrophotometer at wavelength 400 nm in cuvette with a layer thickness 10 mm. As a comparison solution prepared in the same way but without addition of aluminum chloride, is used. In parallel the optical density of the rutin solution-standard sample prepared similarly to testing solution was measured. The concentration of rutin was determined by the optical density data.
The obtained data by the determination of fixing of flavonoids content in extract are presented on
(Fig. 1).
Figure 1. Dependence of optical density on length of wave
Significant variations of the sum of flavonoids are primarily caused by quality ofthe initial plant raw materials. On the base ofobtained data of physico-chemical analysis and the results of pharmacological investigations the content of sum flavonoids in terms on rutin, which was normed and was not less than 0.060%.
Quantitative analysis of micro-and macroelements in the leaves of the plant Asarum europaeum L.with using mass spectrometrw (ICP-MS) with inductively bonded plasma was carried out in the laboratory [5]. 0.05-0.5 g of plant were weighed
on analytical scales and transferred to Teflon autoclaves, after which they are filled with appropriate amount of concentrate mineral acids (nitric acid and hydrogen peroxide). The autoclaves were closed and placed in a microwave oven for decomposition. After decomposition the content in autoclaves are quantitatively transferred in volumetric flasks on 50 or 100 ml and the volume is adjusted to the mark by 0.5% nitric acid. Determination of the test substance is carried out on ISP MS devices or similar device of an optical emission spectrometer with inductively
coupled argon plasma. In the determination method macroelements is indicated at which they have the the optimal wavelength of the determined micro or maximum emission.
Table 1.- The content of elements in Asarum europaeum L.
Element Content, mcg/g Element Content, mcg/g Element Content, mcg/g
Macroelements Selenium (Se) 0.302 Rubidium (Rb) 5.088
Potassium (K) 41526,479 Titanium (Ti) 23.083 Strontium (Sr) 141.038
Calcium (Ca) 28936,645 Vanadium (V) 2.049 Zirconium (Zr) 1.280
Silicon (Si) 259,275 Chromium (Cr) 4.909 Niobium (Nb) 0.079
Magnesium (Mg) 4647,475 Manganese (Mn) 35.618 Molybdenum (Mo) 0.801
Sodium (Na) 593,325 Iron (Fe) 1593.820 Silver (Ag) 3.369
Phosphorus (P) 1439,850 Cobalt (Co) 0.631 Cadmium (Cd) 0.108
Micro and ultramicroelements Nickel (Ni) 2.690 Tin (Sn) 0.574
Lithium (Li) 3,176 Copper (Cu) 8.636 Indium (In) 0.003
Beryllium (Be) 0,108 Zinc (Zn) 38.112 Antimony (Sb) 0.167
Boron (B) 16,901 Gallium (Ga) 3.931 Cesium (Cs) 0.123
Aluminum (Al) 917,141 Germanium (Ge) 0.016 Barium (Ba) 155.165
Sulfur (S) 10.373 Arsenic (As) 0.693 Tantalum (Ta) 0.024
Tungsten (W) 0.013 Rhenium (Re) 0.013 Mercury (Hg) 0.263
Thallium (Tl) 0.024 Lead (Pb) 5.726 Bismuth (Bi) 0.058
Uranium (U) 0.052
On the base of data presented in the table it is shown that macro -, micro - and ultramicroelements were found in the leaves of Asarum europaeum L., which can be presented in the following rows by the content decreasing: macroelements-K > Ca > Mg > >P > Na > Si; micro and ultra-microelements-Fe > Al > >Ba > Sr > S > Cu > Pb > Rb > Cr > Ga > Ag > Si > >Ni > V > Zn > Mo > As > Co > Sn > Se > Hg > Cs > >Cd = Be > Nb > Bi > U > Ta = Tl = Ge > Re = W > In.
Among the macroelements the most contents are atributelto potassium, calcium, and magnesium; among the trace elements - manganese, barium, zinc. In concentrations from 1 to 10 mcg/g,
B, S, Li, Pb, Rb, Cu, Cr were determined; below 1 (within 0.1180-0.9700 mcg/g) - Be, Co, Cs, As, Sb; below 0.1 (0.0190-0.0760 mcg/g) - Tl, W, U, Re, Ge, Bi; below 0.01 (0.001-0.091 mcg/g) - contain Ag and W.
Such it was determined that place of growth of the plant has intensity influenced on of accumulation of elements. It was determined that leaves of the plant Asarum Europaeum L contain macroelements-potassium, calcium, and magnesium; from micro-and ultramicroelements in its composition there are iron, aluminum, strontium, zinc and manganese in large quantities.
References:
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5. Reference of the doctor. Mineral substances of medicinal plants. (Electronic resource) URL: http: //spravr.ru/mineralnye-veshchestva-lekarstvennyh-rasteniy.html.
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