Научная статья на тему 'ESSENTIAL OIL COMPOSITION OF TWO SPECIES OF SCUTELLARIA AERIAL PARTS'

ESSENTIAL OIL COMPOSITION OF TWO SPECIES OF SCUTELLARIA AERIAL PARTS Текст научной статьи по специальности «Фундаментальная медицина»

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SCUTELLARIA ADENOSTEGIA / SCUTELLARIA COMOSA / ESSENTIAL OIL / GC/MS/FID / ANTIBACTERIAL ACTIVITY

Аннотация научной статьи по фундаментальной медицине, автор научной работы — Karimov Abdurashid Musakhonovich, Bobakulov Khairulla Mamadievich, Ostroushko Yulia Vladimirovna, Botirov Erkin Khozhiakbarovich, Mamadrakhimov Azimjon Akparalievich

The chemical composition of essential oils obtained by hydrodistillation method from two plants of the genus Scutellaria, grown in Uzbekistan and used in folk medicine were comparatively investigated by GC/MS and FID. Overall individually thirty three constituents were identified in both of aerial parts of S. adenostegia and S. comosa essential oils, representing 94.4 and 97.0% of the total, respectively. The main components were determined as acetophenone (24.2%), eugenol (12.3%), caryophyllene oxide (8.9%), and β-caryophyllene (7.0%) in the oil of S. adenostegia. β-Caryophyllene (12.5%), phytol (11.4%), linalool (11.1%), acetophenone (10.4%), caryophyllene oxide (6.6%),1-hexanol (5.3%), and (E)-2-hexenal (5.1%) were found as major components in the S. comosa oil. The composition of the oils of S. adenostegia and S. comosa was being reported for the first time. The essential oils of S. adenostegia and S. comosa showed significant antimicrobial properties against Bacillus subtilis, moderate effect against Salmonella enterica and Escherichia coli.

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Текст научной работы на тему «ESSENTIAL OIL COMPOSITION OF TWO SPECIES OF SCUTELLARIA AERIAL PARTS»

XHMHfl PACTfflE.nbHOro CblPka. 2021. №4. C. 139-144.

DOI: 10.14258/jcprm.2021049121

UDC 547.913:543.544.32

ESSENTIAL OIL COMPOSITION OF TWO SPECIES OF SCUTELLARIA AERIAL PARTS

© A.M. Karimov1, Kh.M. Bobakulov1'2, Yu.V. Ostroushko3, E.Kh. Botirov3', A.A. Mamadrahimov4, N.D. Abdullaev1

1 Acad. S.Yu. Yunusov Institute of the Chemistry of Plant Substances, Academy of Sciences of the Republic of Uzbekistan, ul. M. Ulugbeka, 77, Tashkent, 100170 (Uzbekistan)

2 Tashkent Institute of Irrigation and Agricultural Mechanization Engineers, ul. Kori Niyoziy, 39, Tashkent, 100000 (Uzbekistan)

3 Surgut State University, ul. Lenina, 1, Surgut, 628412 (Russia), e-mail: botirov-nepi@mail.ru

4 Institute of Bioorganic Chemistry, UzAS, ul. M. Ulugbeka, 83, Tashkent, 100125 (Uzbekistan)

The chemical composition of essential oils obtained by hydrodistillation method from two plants of the genus Scutellaria, grown in Uzbekistan and used in folk medicine were comparatively investigated by GC/MS and FID. Overall individually thirty three constituents were identified in both of aerial parts of S. adenostegia and S. comosa essential oils, representing 94.4 and 97.0% of the total, respectively. The main components were determined as acetophenone (24.2%), eugenol (12.3%), caryo-phyllene oxide (8.9%), and ¿8-caryophyllene (7.0%) in the oil of S. adenostegia. ¿8-Caryophyllene (12.5%), phytol (11.4%), lin-alool (11.1%), acetophenone (10.4%), caryophyllene oxide (6.6%),1-hexanol (5.3%), and (E)-2-hexenal (5.1%) were found as major components in the S. comosa oil. The composition of the oils of S. adenostegia and S. comosa was being reported for the first time. The essential oils of S. adenostegia and S. comosa showed significant antimicrobial properties against Bacillus subtilis, moderate effect against Salmonella enterica and Escherichia coli.

Keywords: Scutellaria adenostegia; Scutellaria comosa; essential oil; GC/MS/FID; antibacterial activity.

This work was supported by the Program for Fundamental Scientific Research of the Uzbekistan Academy

of Sciences under the Grant VA-FA-F6-010.

Introduction

The genus of Scutellaria L., skullcaps, "Ko'kamaron" (local name) which belongs to the family Lamiaceae, is represented by 360 species and is widely spread in mild, subtropical, and tropical regions of the world, including Eu-

Karimov Abdurashid Musakhonovich - PhD (Chemistry), Senior Researcher, Laboratory of Chemistry of coumarins and terpenoids, e-mail: abdurashidka@mail.ru Bobakulov Khairulla Mamadievich - Candidate of Chemical Sciences, Senior Researcher, Laboratory of Physical Research Methods, e-mail: khayrulla@rambler.ru Ostroushko Yulia Vladimirovna - postgraduate student, e-mail: julliyaost@gmail.com Botirov Erkin Khozhiakbarovich - Professor of the Department of Chemistry, e-mail: botirov-nepi@mail.ru Mamadrakhimov Azimjon Akparalievich - Candidate of Chemical Sciences, Senior Researcher, Laboratory of Complex Compounds, e-mail: kimyogar@yahoo.com Abdullaev Nasrulla Jalilovich - Professor, leading researcher, Laboratory of Physical Methods of Research, e-mail: n_abdullaev@rambler.ru

rope, North America, and Eastern Asia [1]. Approximately 120 species and subspecies of the genus grow across the countries of Commonwealth of Independent States (CIS), mainly in the Caucasus Mountains and in Middle Asia [1, 2]. Skullcaps are perennial or, very rarely, annual grasses, rarely subshrubs or half-shrubs. Many of the skullcap species are decorative plants, some are medicinal herbs, but all belong to dyeing plants. In Uzbekistan, there are thirty eight species of the Scutellaria L. genus plants. It is used in Uzbekistan in treatment of inflammation, chorea, nervous tension and high-blood pressure [3]. Chemical composition of

* Corresponding author.

plants of the Scutellaria L. genus is diverse. Earlier, flavonoids, phenylpropanoids, phenolic acids, iridoids, clerodane diterpenoids, steroids, triterpenes, lignans, alkaloids, phytosterols, polysaccharides, tannin substances, essential oils, and other classes of natural compounds have been isolated from different species [1, 2, 4]. S. adenostegia Briq. is a perennial native plant growing on rocky and clay mountain slopes, dried up riverbeds and streams, rocky placers and gravels along the banks of the rivers of Tian-Shan, Pamir-Alay Mountains (Central Asia). Previous phytochemical studies on this plant reported the isolation and identification of flavonoids [3]. S. comosa Juz. mainly occurring in the Tian Shan and Pamir-Alai Mountains, it is a perennial shrub species which endemic to the Central Asia. Flavonoids on this plant species have been extensively studied [3, 5]. Several authors reported on the study of essential oils of plants of Scutellaria genus [1, 6-10].

It was previously reported that essential oils isolated from plants of this genus possess antioxidant and antibacterial activities [10-14]. In the literature, there is a report on the study of the component composition and antioxidant activity of essential oils of three species of plants of the genus Scutellaria, growing in Uzbekistan [10]. But until now, there are no published reports concerning the phytochemistry and biological activities of the essential oils of S. adenostegia and S. comosa. We have reported here the isolation and characterization of essential oils, which to the best of our knowledge, is the first investigation on volatile compositions of the aerial parts of two Scutellaria species.

Experimental

The aerial parts (stems, leaves, flowers) of S. adenostegia and S. comosa employed in this investigation were collected in the flowering stage (May, 2019) from Chust (41°00'00" N 71°13'59.88" E) and Turakurgan District (41°00'00" N 71°30'56.88" E) Namangan Region of Uzbekistan respectively. The plants were identified at the Flora of Uzbekistan Department, Institute of Botany (Uzbekistan) by Dr. O.T. Turginov. The voucher specimens of S. adenostegia (accession number (A.N.) N20190550) and S. comosa (A.N. N20190551) have been deposited at the Flora of Uzbekistan Department. Isolation of the essential oil. The air-dried aerial parts (moisture content was 1113% w.b.) of the S. adenostegia and S. comosa were hydrodistilled three times (3x100 g each) for 3 h, using a Clevenger-type apparatus. Further hydrodistillation of plant raw materials did not lead to an increase in the yield of essential oil. The obtained essential oils were then dried using anhydrous sodium sulfate and stored at 4 °C in the dark until use. The essential oil content was calculated as a relative percentage (v/w) of the dry plant material.

GC and GC/MS analysis of essential oils. The qualitative and quantitative composition of the essential oils were determined on an Agilent 5977B MSD/8890A GC (Agilent Technologies, USA) gas chromatography-mass spectrometer equipped with flame ionization detector (FID) and an Agilent 7693A ALS autoinjector. The components of the mixture were separated on an Agilent DB-Wax quartz capillary column (30 m x 250 |im x 0.25 |im film thickness) in the following temperature mode: 50 °C (1 min) - 4 °C/min to 200 °C (6 min) - 15 °C/min to 250 °C (35 min). The samples were prepared in dichloromethane and 1.0 |L injected in splitting mode (50 : 1). The flow rate of the mobile phase (H2) was 1.1 mL/min. The injector temperature was 220 °C. MS conditions were as follows: ionization energy 70 eV, source temperature 230 °C, quadrupole temperature 150 °C. EI-MS spectra were obtained in the m/z range of 10-550 a.m.u.

FID was used for the quantification of the volatile compounds. The chromatographic conditions and the column were identical to those used for the GC/MS analysis. The injector temperature was 250 °C and the carrier gas was H2 at 1.1 mL/min.

Components of essential oils were identified by comparison of the chromatographic peaks retention times with those of authentic compounds analyzed under the same conditions, and by comparison of retention indices (as Kovats indices) with literature data [15].

Comparisons of MS fragmentation patterns with mass spectrum database search were performed using the Wiley Registry of Mass Spectral Data-9th Ed., NIST Mass Spectral Library (2011) and the Automated Mass Spectral Deconvolution and Identification System (AMDIS, Version 2.72) containing NIST14 Library. A C9-C32 n-alkane standard solution (Agilent Technoligies, USA) was used for the determination of chromatographic retention indices (RI). Percent composition was obtained for each constituent on the basis of flame ionization detection analyses of the essential oils.

Antimicrobial activity. The essential oils antimicrobial activities were assessed against four pathogenic bacterial strains, two Gram-positive Staphylococcus aureus (MTCC 737) and Bacillus subtilis (NK-1, isolated from

Natto) and two Gram-negative Salmonella enterica (ATCC 14028) and Escherichia coli (MTCC 1302). The above microorganisms were obtained from the Department of Microbiology of the Medical Institute of Surgut State University. The assay was performed using a 96-well microliter plate-based method with resazurin as a cell growth indicator following a previously used method with minor adjustments [16].

Statistical analysis. Results were expressed as mean + standard deviation. Statistical comparisons were performed with Student's t-test using GraphPad Prism version 7.00. Differences were considered significant at p <0.05.

Results and discussion

Chemical composition of essential oils. The average yields of essential oils obtained from three independent determinations by the hydrodistillation method were 0.19% (v/w; S. adenostegia; light yellow), and 0.17% (v/w; S. comosa; yellow) on a dry weight. The chemical compositions of the essential oils isolated from the air-dried aerial parts of plants were investigated by GC/MS/FID. Chromatographic profile of the volatiles from two Scutellaria species on the DB-Wax column is presented on Figure and Table 1 show the composition of compounds identified from the studied oil samples. Totally thirty three compounds representing 94.4% of the total oil were characterized in essential oil of S. adenostegia. The classes of compounds present in S. adenostegia were aldehydes and ketones (35.2%), phenols (16.0%), alcohols (12.4%), sesquiterpene hydrocarbons (12.3%) and oxygenated sesquiterpenes (11.0%).

The main compounds of S. adenostegia were acetophenone (24.2%), eugenol (12.3%), caryophyllene oxide (8.9%), and ^-caryophyllene (7.0%). Moreover, high amount of 1-hexanol (3.8%), furfural (3.3%) and y-himachalene (2.7%) were also observed in essential oil. It should be noted that acetophenone was also the main component of the essential oils of S. immaculate and S. schachristanica [10]. The total phenolic compounds content in essential oil of this plant was 13.3%, of which 11.8% was eugenol. S. adenostegia essential oil were rather poor in content of oxygenated monoterpenes (3.0%).

Chromatographic profile of the volatiles from essential oils of Scutellaria adenostegia Briq. (upper) and

Scutellaria comosa Juz. (lower)

Table 1. Essential oil composition of two Scutellaria species

No Compounds RT, min LRI* S. adenostegia, S. comosa,

(Conc., %) (Conc., %)

1 1,8-Cineol 9.231 1192 - 0.9

2 (£)-2-Hexenal 9.495 1202 1.6 5.1

3 1-Hexanol 13.466 1341 3.8 5.3

4 (Z)-3-Hexen-1-ol, 14.364 1369 - 2.0

5 1-Octen-3-ol 16.315 1433 - 3.9

6 Furfural 16.584 1442 3.3 -

7 a-Cubebene 17.465 1472 0.7 0.8

8 Pentadecane 17.637 1500 - 1.4

9 Camphor 18.158 1516 - 1.7

10 Benzaldehyde 18.301 1520 1.8 1.1

11 Linalool 19.142 1549 1.1 11.1

12 1-Octanol 19.491 1560 1.9 1.5

13 (,E)-S-Caryophyllene 20.475 1593 7.0 12.5

14 Acetophenone 21.923 1624 24.2 10.4

15 a-Caryophyllene 22.461 1644 1.1 1.7

16 a-Terpineol 23.319 1676 0.6 1.2

17 y-Himachalene 23.886 1697 2.7 2.4

18 5-Cadinene 24.841 1733 0.8 0.5

19 Grandlure II 27.290 1829 - 0.7

20 (Z)-Geranylacetone 27.387 1833 0.5 -

21 Benzyl alcohol 27.960 1856 2.1 1.3

22 2-Phenylethanol 28.835 1892 1.2 0.7

23 (£')-yS-Ionone 29.476 1918 0.4 0.5

24 Caryophyllene oxide 30.546 1961 8.9 6.6

25 Alloaromadendrene oxide-(1) 30.866 1974 - 0.5

26 o-Cresol 31.038 1981 1.6 -

27 (Z)-Nerolidol 31.845 2015 0.6 -

28 4-Phenyl-3-buten-2-one 33.573 2092 0.4 -

29 Hexahydrofarnesyl acetone 33.882 2120 3.0 2.1

30 Eugenol 34.769 2145 12.3 1.3

31 a-Muurolol 35.101 2159 0.6 0.9

32 9yS-Acetoxy-3,4,8-trimethyltricyclo[6.3.1.0(1,5)]dodec-3-ene 35.272 2167 - 0.8

33 ^-Vinyl-guaiacol 35.393 2172 1.3 0.6

34 Caryophylla-4(12),8(13 )-dien-5a-ol 37.647 2272 1.5 1.2

35 Dihydroactinidiolide 38.471 2309 1.3 1.8

36 8-Cedren-13-ol 39.335 2347 - 0.9

37 2,3-Dihydro-benzofuran 39.650 2361 2.1 -

38 Coumarin 40.863 2416 0.7 -

39 Pentacosane 42.528 2500 0.8 1.1

40 Phytol 45.601 2643 2.8 11.4

41 Acetovanillone 46.253 2675 0.8 -

42 Methoxyacetic acid 2-pentadecyl ester 47.260 2723 0.9 1.1

Oxygenated monoterpenes - 1, 9, 11, 16, 19, 35. 3.0 17.4

Sesquiterpene hydrocarbons - 7, 13, 15, 17, 18. 12.3 17.9

Oxygenated sesquiterpenes - 24, 25, 31, 32, 34, 36. 11.0 10.9

Aldehydes and ketones - 2, 6, 10, 14, 20, 23, 28, 29. 35.2 19.2

Alcohols - 3, 4, 5, 12, 21, 22, 27, 40. 12.4 26.1

Phenols - 26, 30, 33, 41. 16.0 1.9

Others - 8, 37, 38, 39, 42 4.5 3.6

Total 94.4 97.0

Identified compounds 33 33

*LRI - Linear retention indices on DB-Wax column; Conc., % calculated from FID data.

In the essential oil of S. comosa thirty five components, representing 97.0% of the total ones were characterized (Tab. 1). In the studied essential oil alcohols (26.1%) dominated. Aldehydes and ketones (19.2%), sesquiterpene

hydrocarbons (17.9%), oxygenated monoterpenes (17.4%) and oxygenated sesquiterpenes (10.9%) were the remaining groups of components (Tab. 1).

¿^-Caryophyllene (12.5%), phytol (11.4%), linalool (11.1%), acetophenone (10.4%), caryophyllene oxide (6.6%), 1-hexanol (5.3%), and (E)-2-hexenal (5.1%) were found as the main constituents. Literature information indicated that ¿^-caryophyllene was the main component of the essential oils of S. brevibracteata [6], S. sibthorpii [7], S. luteo-caerulea [9], S. albida [12] and other species. Acetophenone was the main component of S. schachristanica and S. immaculata essential oil [10], while linalool was determined as a major component for S. cypria var. elatior [7]. In total, forty four volatile compounds were identified in two Scutellaria species from Uzbekistan. Aldehydes and ketones, alcohols, phenols and sesquiterpene hydrocarbons are the dominant components of the essential oil of the plant S. adenostegia. Sesquiterpene hydrocarbons and oxygenated monoterpenes were the major group of terpenes found in the essential oil of S. comosa. On the contrary, monoterpene hydrocarbons were not detected in both oil of S. adenostegia and S. comosa.

Evaluation of antibacterial activity. The literature contains data on the antimicrobial activity of essential oils of some plants of the Scutellaria genus [7, 11-14]. That is, we have studied the antibacterial properties of the isolated essential oils. The essential oils of S. adenostegia and S. comosa showed significant antimicrobial properties against Bacillus subtilis (318.0±8.62 and 401.1±14.49 ^g/mL), moderate effect against Salmonella enterica (519.4±16.29 and 803.1±31.62 ^g/mL) and Escherichia coli (528.3±14.63 and 802.4±32.57 ^g/mL), and weak effect against Staphylococcus aureus (1297.6±34.78 and 1676.3±52.94 ^g/mL) respectively (Table 2). We are inclined to believe that the antibacterial activity of essential oils is due to the presence of ^-caryophyllene and eugenol in their composition [17, 18]. Pure ¿^-caryophyllene showed more pronounced antibacterial activity against Gram-positive bacteria than Gram-negative bacteria [17]. Overall, the antibacterial activity of the essential oils can be related to the content of many of the compounds identified in the oils, including eugenol and ¿^-caryophyllene.

Table 2. Minimum inhibition concentration (MIC) of essential oils of S. adenostegia and S. comosa against four bacterial strains

Samples MIC (^g/mL)a

E. coli S. aureus B. substilis S. enterica

Streptomycinb 25.8±1.23 3.1±0.12 25.8±1.07 25.8±1.28

S. adenostegia. 528.3±14.63 1297.6±34.78 318.0±8.62 519.4±16.29

S.comosa. 802.4±32.57 1676.3±52.94 401.1±14.49 803.1±31.62

a All results are presented as mean±standard deviations for triplicate assays.

b Reference. Conclusions

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For the first time, the chemical composition of the essential oils grown in Uzbekistan of two Scutellaria species was studied. Acetophenone and ¿^-caryophyllene are the dominant terpenes of the essential oils of S. adenostegia and S. comosa. On the other hand the high amount of eugenol and caryophyllene oxide in essential oil of S. adenostegia and linalool, phytol and caryophyllene oxide in essential oil of S. comosa were revealed. The essential oils of these plants showed significant antibacterial properties against Bacillus subtilis, moderate effect against Salmonella enterica and Escherichia coli.

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Received January 4, 2021 Revised April 25, 2021 Accepted June 28, 2021

For citing: Karimov A.M., Bobakulov Kh.M., Ostroushko Yu.V., Botirov E.Kh., Mamadrahimov A.A., Abdullaev N.D. Khimiya Rastitel'nogo Syr'ya, 2021, no. 4, pp. 139-144. (in Russ.). DOI: 10.14258/jcprm.2021049121.

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