Chromato-mass-spectroscopic research of chemical composition of Elaeagnus angustifolia L.

The aim of the work was chromato-mass-spectroscopic research of chemical composition of Elaeagnus angustifolia L. fruits and leaves and identifying further prospects for the use of this plant in medicine.
Materials and methods. Raw materials of Elaeagnus angustifolia L. were selected as objects of study. The tincture was obtained by maceration and the raw material was extracted with methyl alcohol at room temperature for 10 days according to the method of making tinctures. The study of the chemical composition of Elaeagnus angustifolia L. was carried out using gas chromatograph Agilent 7890B GC System (Agilent, Santa Clara, CA, USA) with mass spectrometric detector Agilent 5977 BGC/MSD (Agilent, Santa Clara, CA, USA) and chromatographic column DB-5ms (30 m × 250 mkm × 0,25 mkm).
Results. 23 compounds (1 in the isomeric state) in fruits and 20 compounds (2 in the isomeric state) in the leaf of Elaeagnus angustifolia L. were identified. The main components of fruits were sitosterol (phytosterols) – 12.53 %, propyl acetate (esters of carboxylic acids) – 12.60 %, chamazulene (terpenes) – 11.97 % and palmitic acid (fatty acids) – 8.28 %. The main component of leaves were sitosterol (phytosterols) – 17.57 %, 1-(2-hydroxy-5-methylphenyl)-ethanone (ketone) – 8.35 %, phytol (terpenes) – 6.10 %.
It is known from the literature that chamazulene has antioxidant, antinociceptive, cytotoxic activity. Sitosterol has anti-inflammatory and antidiabetic activity. Hexadecanoic (palmitic) acid has antimicrobial, antidiabetic and antioxidant properties. Phytol is characterized by a wide range of biological action – antimicrobial, antinociceptive, anti-inflammatory, antioxidant and cytotoxic.
Conclusions. Based on the above, the olive can be considered as a source of antimicrobial, antinociceptive, anti-inflammatory, antioxidant, antidiabetic and cytotoxic drugs.

Актуальные вопросы фармацевтической и медицинской науки и практики. 2021. Т. 14, № 2(36). С. 179-184 The family Eleagnaceae includes about 80 species of plants [1]. In forms of life, these are deciduous trees and shrubs, less often vines, which grow mainly in Western Europe, Asia Minor, Central, and Southeast Asia, the Far East, and North America [2]. These plants play a very important ecological role and are used in folk medicine in some eastern countries [3]. Of note is the narrow-leaved olive Elaeagnus angustifolia L. Thus, Iranian researchers have proven the effectiveness of aqueous extract of olive fruit in the treatment of osteoarthritis [4,5]. Scientists from the Tehran University of Medical Sciences have described the cardioprotective, antioxidant, analgesic, antitumor, and anti-inflammatory effects of olive extracts [6]. These data are confirmed by studies of a team of American and Iranian scientists [7]. Other Iranian scientists have studied the antioxidant effect and considered the possibility of using olives as a biological additive of probiotic nature [8].
However, it was not revealed sufficient information on the chemical composition of Elaeagnus angustifolia L. in the literature. Therefore, research in this area is expedient and relevant.

Aim
The purpose of our work is to study the chemical composition of olives of narrow-leaved fruits and leaves by chromato-mass spectroscopic and to catch out on further promising applications of this plant in medicine.

Materials and methods
The object of the study was fruits and leaves of narrow-leaved olives. Raw materials were collected in August 2020 during the flowering and fruiting phase (Zaporizhzhia, Vyrva tract).
The tincture was obtained by maceration, and the raw material was extracted with methyl alcohol at room temperature for 10 days according to the method of making tinctures. 0.1 ml of the extract was placed in a 1 ml micro flask and made up to 0.5 ml with methanol [9].
Qualitative and quantitative determination of active compounds was carried out at the Department of Natural Sciences for Foreign Students and Toxicological Chemistry (Head of the Department -PhD, DSc, Professor O. I. Panasenko).
The completeness of the reactions and the individuality of the resulting compounds were controlled by the gas chromatograph Agilent 7890B with a 5977B mass spectrometry detector. The column is DB-5ms 30 m × 250 μm × 0.25 μm with length. The gas-carrier speed (helium) is 1.6 ml/min. Injection volume -0.5 μl. Separation of the flow is 1:50. The temperature of the sampling unit is 230 °C → 12 °C/s → 275 °C. Thermostat temperature: programmable, 240 °C (1 minute delay) → 5 °C/ min → 280 °C (delay 1 min.). The total time of examination is 10 min. Temperature of interface GS/MS -280 °C; ion sources -230 °C; quadrupole mass analyzer -150 °C. Type of ionization: EI with an electron energy of 70 eV. The range of mass numbers that was scanned: 30-500 m/z.

Results
According to the results of the study, 23 compounds (1 in the isomeric state) in fruits and 20 compounds (2 in the isomeric state) were identified in the leaf of Olive narrow-leaved (Tables 1, 2), which by chemical structure belong to different classes of organic substances. The largest part in the chemical composition of fruits, was esters of carboxylic acids (5 com-pounds, 24.32 %), heterocyclic compounds (5 compounds, 14.53 %), phytosterols (1 compound, 12.53 %), terpenes (1 compound, 11.97 %).

Discussion
According to the literature chamazulene has antioxidant [10] and antinociceptive activity [11]. Italian scientists have studied its effects on human melanoma cells, in which it was found that plants rich in chamazulene were particularly active against cancer cells, causing their death [12].
Sitosterol is characterized by anti-inflammatory activity [13,14]. Japanese scientists have examined its ability to inhibit chronic inflammation associated with obesity [15]. Based on the data obtained, it was concluded that sitosterol can be used to treat metabolic diseases, in particular diabetes [16].
Hexadecanoic (palmitic) acid has antimicrobial [17], antioxidant properties and can also be used in the treatment of diabetes [18].
Phytol is characterized by a wide range of biological activity, that is antimicrobial, antinociceptive, anti-inflammatory, antioxidant, and cytotoxic [19,20].
Based on the research, the olive can be considered as a source of antimicrobial, antinociceptive, anti-inflammatory, antioxidant, and cytotoxic medicines. Also, it can be considered useful in the treatment of diabetes.

Conclusions
1. For the first time the chemical composition of olive-leaved fruits and leaves was established with GS/MS.
2. According to the results of the study, 23 compounds (1 in the isomeric state) in fruits and 20 compounds (2 in the isomeric state) were identified in the leaf of Olive narrow-leaved, which by chemical structure belong to different classes of organic substances.
4. Olive narrow-leaved can be recommended for further research on antimicrobial, antinociceptive, anti-inflammatory, antioxidant, and cytotoxic, antidiabetic activity.
Prospects for further research. Olive narrow-leaved fruits and leaves can be considered as a potential source of antimicrobial, antinociceptive, anti-inflammatory, antioxidant, cytotoxic, and antidiabetic drugs.

Funding
The work is part of the joint integrated work of the Department of Pharmacognosy, Pharmacology and Botany of Zaporizhzhia State Medical University.