CHEMICAL COMPONENTS AND VARIABILITY OF THE ESSENTIAL OILS ISOLATED FROM INFLORESCENCES OF CARDUUS SPP.

Transcription

1 Acta Poloniae Pharmaceutica ñ Drug Research, Vol. 73 No. 5 pp. 1405ñ1411, 2016 ISSN Polish Pharmaceutical Society CHEMICAL COMPONENTS AND VARIABILITY OF THE ESSENTIAL OILS ISOLATED FROM INFLORESCENCES OF CARDUUS SPP. MA GORZATA KOZYRA 1 * and MAREK MARDAROWICZ 2 1 Department of Pharmacognosy with Medicinal Plant Laboratory, Medical University of Lublin, 1 Chodüki St., Lublin, Poland 2 Analytical Laboratory, Maria Curie-Sklodowska University of Lublin, Poland Keywords: Carduus species, essential oil, GC/MS method Carduus is a predominantly Eurasian genus of about 90 species that belongs to the Asteraceae family (1). That plant contains multiple secondary metabolites: flavonoids, phenolic acids, alkaloids, sterols, triterpenes, polyacetylenes, acetylenes, sterols hydrocarbons, coumarins and lignans. (1-6) There are only a few information about essential oils in Carduus sp. The prominent component found in essential oils obtained from aerial parts of C. pycnocephalus L. was palmitic acid (39.62%). In GC/MS analysis of the petroleum ether extract of the aerial parts of the plant sixteen compounds were identified. Olean-12-en-3-α-ol (20.39%), ursa-9(11),12- dien-3-ol (17.74%) and palmitic acid (17.62%) were the major components (7). Another authors identified 29 components in the oil of C. pycnocephalus representing 83.4% of the total oil with hexadecanoic acid (23.3%) as a main constituent (8). The major essential oil components from flowers of C. candicans ssp. globifer were benzaldehyde (22.1%), palmitic acid (8.9%), methyl salicylate (7%), heptacosane (6%), tricosane (6.1%), pentacosane (5%), Z-12-pentacosene (3%) and β- caryophyllene (3%). Palmitic acid (17.9%) together with methyl salicylate (14%), benzaldehyde (13.2%), trans-nerolidol (4%), p-cymen-8-ol (4%) and tricosane (2%) were detected as most characteristic compounds occurring in the essential oils from C. thoermeri (9). Essential oils received from aerial parts of C. nutans growing in Italy, were investigated and the major component of this essential oils was hexadecanoic acid (18.6%), hexahydrofarnesylacetone (7.8%), heptacosane (5.9%), 4-vinyl guaiacol (5.8%), pentacosane (3.8%) and eugenol (3.6%). (1). The aim of these studies was qualitative analysis of essential oils hydrodistilled from the inflorescences of three Carduus species growing in Poland. These were: Carduus crispus L., C. defloratus L. and C. nigrescens Vill. Essential oils from the inflorescences of investigated species have not been examined so far. MATERIALS AND METHODS Plant material The inflorescence of Carduus L. species were collected in the Medicinal Plant Garden, of the Department of Pharmacognosy, Lublin, Poland. The inflorescence were dried in air at room temperature and powdered according to the accepted normal procedures. The procedure of preparation followed the conditions described in the Polish Pharmacopoeia VI. Voucher specimens were given to the plants and the samples were deposited in the herbarium of The Department of Pharmacognosy, Medical University, Lublin. Hydrodistillation Air dried and powdered inflorescence of Carduus L. spp. (10.0 g) were distilled in glass Deryngís apparatus for 3 h to obtain yellowish oils according to the parameters presented in the Polish Pharmacopoeia VI (10). In this method, three-hour hydrodistilations of the plants material with water vapor with the addition of m-xylene is applied. The above methods allowed to obtain samples of essential oils from Carduus spp. The samples were * Corresponding author: mkozyra@pharmacognosy.org; phone: ,

2 1406 MA GORZATA KOZYRA and MAREK MARDAROWICZ Table 1. The composition of the examined essential oils from inflorescences Carduus crispus L., Carduus defloratus L., Carduus nigrescens Vill. (hydrodistillation with m-xylene) determined by GC/MS. Compounds RI* Carduus spp. (Ref. no.) name RI C. crispus C. defloratus C. nigrescens Peak area (%) 2-Pentyl-furan Linalool Nonanal Terpinen-4-ol p-cymen-8-ol α-terpineol Methyl salicylate n-decanal Thymol methyl ether Thymol Carvacrol Eugenol (E)-fl-Damascenone β-cubebene Tetradecane fl-caryophyllene α-humulene E-fl-Farnezane E-fl-Ionone fl-selinen Tridecanal (E)-Nerolidiol Dodecanoic acid Hexadecane Humulene epoxide Tetradecanal α- Cadinol α-bisabolol Unknown

3 Chemical components and variability of the Carduus spp Table 1. Cont. Compounds RI* Carduus spp. (Ref. no.) name RI C. crispus C. defloratus C. nigrescens Peak area (%) Pentadecanal Tetradecanoid acid methyl ester Unknown Tetradecanoic acid Octadecane Hexadecanal Pentadecanone Nonadecane Heptadecan-2-one Heptadecanal 1923 NIST Palmitic acid methyl ester Z-11-Hexadecenoic acid Palmitic acid Unknown compound 2081a Linoleic acid methyl ester (14) Linolenic acid methyl ester Docosane Tricosane Tetracosane Pentacosene isomer I 2475 NIST Pentacosene isomer II 2483 NIST n-pentacosane n-hexacosane n-heptacosane n-octadecane Squalene 2833 NIST n-nonacosane ,8 Tricosane Hentriacontane RI - retention indices, * retention indices from the literature data. NIST - National Institute of Standards and Technology, Gaithersburg, MD, USA. placed in small glass vials, dried over anhydrous sodium sulfate, and stored at 4 C until further analysis. Gas chromatography-mass spectrometry analysis GC-MS method was employed for the analysis of obtained essential oils. Analysis was performed by use of Thermo-Finnigan (USA) GCQ GC-MS apparatus, working in electron impact mode. The extracts were separated on a 20 m 0.18 mm i.d. capillary column coated with a 0.2 µm film of RT-5 (Restek). The volume of sample injected was 1 µl and split injection was used (split ratio 1 : 50).

4 1408 MA GORZATA KOZYRA and MAREK MARDAROWICZ Helium was used as carrier gas at a flow rate of 0.5 ml/min. The analyses were carried out in the programmed mode with temperature gradient of O C at 4 O C/min. The final temperature was held for 1 min. The mass spectrometer in the electron impact (EI) ionization mode (70 ev), and full-scan mass spectra were recorded in the range m/z a.m.u. (11). The identification of individual compounds was based on the calculated retention indices, as well as was made by the comparison of received mass spectra with those of reference compounds, and available in NIST (National Institute of Standards and Technology, Gaithersburg, MD, USA) library, MS data from the literature (12-16) and our library databases. RESULTS AND DISCUSSION Volatile compounds identified by GC/MS method are presented in Table 1. A gas chromatograms of the essential oils from Carduus spp. are presented in Figures 1-3. As it is seen there, compounds were separated and identified from the essential oils examined. Table 1 presents in detail the results of the analysis. All described compounds were identified on the basis of the comparison of their mass spectra with Figure 1. GC chromatogram of essentials oil (hydrodistillation with m-xylene) obtained from inflorescences of Carduus crispus L. The rate of temperature programming 4 O C/min Figure 2. GC chromatogram of essentials oil (hydrodistillation with m-xylene) obtained from inflorescences of Carduus defloratus L. The rate of temperature programming 4 O C/min

5 Chemical components and variability of the Carduus spp Figure 3. GC chromatogram of essentials oil (hydrodistillation with m-xylene) obtained from inflorescences of Carduus nigrescens Vill. The rate of temperature programming 4 O C/min Figure 4. MS spectrum of an unknown compound with RI-1711 from essentials oil obtained from inflorescences Carduus L. spp. Figure 5. MS spectrum of an unknown compound with RI-1732 from essentials oil obtained from inflorescences Carduus L. spp.

6 1410 MA GORZATA KOZYRA and MAREK MARDAROWICZ Figure 6. MS spectrum of an unknown compound with RI 2081a from essentials oil obtained from inflorescences Carduus L. spp. the data available in libraries (see Materials and Methods). The main components in volatile oil obtained from inflorescence from investigated plants are fatty acid - palmitic acids in C. crispus (38.7%), C. nigrescens (31.9%), C. defloratus (30.9%). Another fatty acids with odd number of carbon (linoleic myristic, lauric, linolenic and their derivatives) were also observed. Volatile oils contains aliphatic hydrocarbons with a long carbon side chain. Small amounts of terpenes were also present. Thymol is represented in C. crispus (6.8%), C. defloratus (3.4%), C. nigrescens (0.3%). Carvacrol was identified in C. crispus and C. defloratus (0.3%). Eugenol was represented only in C. nigrescens (0.6%). Linalool was present in inflorescence of C. defloratus (0.3%), and C. nigrescens (0.5%). β-caryophyllene is represented in C. crispus (1.3%) and in C. defloratus (2.8%). Small amount of α-bisabolol was identified only in C. nigrescens (0.5%). In essential oil of C. crispus 38 compounds were analyzed: palmitic acid (38.7%), thymol (6.8%), n-heptacosane (5.7%), linolenic acid metyl ester (4.2%),myristic acid (2.7%), n- pentacosane (2.7%), n-nonacosane (2.6%), linoleic acid (1.4%). In essential oil of C. defloratus 49 compounds were found: palmitic acid (30.9%), myristic acid (6.7%), n- heptacosane (6.7%), thymol (3.4%), n-pentacosane (3.3%), β-caryophyllene (2.8%), n- nonacosane (2.9%). In essential oil of C. nigrescens 44 compounds were analyzed: methyl salicylate (2.3%), myristic acid (9.4%), palmitic acid (31.9%), lauric acid (6.45%), tricosane (3.8%), n-heptacosane (3.6%), n-pentacosane (2.5%). CONCLUSION The aim of this studies were phytochemical investigations of essential oils from Carduus spp. inflorescence by GCñMS method. Compounds of essential oils obtained from inflorescence Carduus species have been identified for the first time. Essential oils were obtained by hydrodistillation in the Deryngís apparatus. All identified compounds were identified on the basis of the comparison of their mass spectra with the data available in libraries. Most of them are ketones and aldehydes with a long carbon side chain. Our study shows the differences in chemical composition of volatile oils obtained from different Carduus spp. The major component of the volatile oil received from the investigated inflorescence was palmitic acid. The percentage of this compound in the essential oils of C. crispus was 38.7%, for C. nigrescens 31.9%, and for C. defloratus 30.9%. Another fatty acids with odd number of carbon (linoleic, myristic, lauric, linolenic and their derivatives) were also observed. In the investigated essential oils the presence of small quantities of terpenoids were also detected These were: thymol, linalool, carvacrol, and β-caryophyllene, among others. In C. nigrescens α-bisabolol was identified (0.5%). Three further unknown compounds with retention indices 1711, 1732, 2081a were also found.

7 Chemical components and variability of the Carduus spp Carduus sp. are sources of bioactive compounds such as thymol, carvacrol, β-caryophyllene and linalool. Declaration of interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. REFERENCES 1. Formisano C., Rigano D., Senatore F., Feo V., Bruno M., Rosselli S.: J. Plant Interact. 2, 115 (2007). 2. Harborne J.B.: in The Biology and Chemistry of the Compositae. Heywood V.H., Harborne J.B., Turner B.L., Eds., p. 359, Academic Press, London Jordon-Thaden I.E., Louda S.M.: Biochem. Syst. Ecol. 31, 1353 (2003). 4. Kozyra M., G owniak K., Boguszewska M.: Curr. Issues Pharm. Med. Sci. 26, 10 (2013). 5. Mabry T.J., Markham K.R., Thomas M.B.: The Systematic Identification of Flavonoids. Springer-Verlag, New York Zhang Q.Y., Wang X.Y., Ying H.P., Cheng T.M., Zhao Y.Y.: Zhongguo Zhongyao Zazhi, 26, 837 (2001). 7. Esmaeili A., Rustaiyan A., Nadimi M., Masoudi S., Tadayon F. et al.: J. Essent. Oil Res. 17, 539 (2005). 8. Al-Shammari L.A., Hassan W.H.B., Al- Youssef H.M: J. Chem. Pharm. Res. 4, 1281 (2012). 9. Zhelev I., Dimitrova-Dyulgerova I., Merdzhanov P., Stoyanova A.J.: Chem. Pharm. Res. 17, 196 (2014). 10. Polish Pharmacopoeia VI. Polish Pharmaceutical Society, Warsaw Kozyra M., Mardarowicz M., KochmaÒska J.: Nat. Prod. Res. 29, 1941 (2015). 12. Adams R.P.: Identification of essential oil components by gas chromatography/mass spectrometry. Allured Publishing Co., Carol Stream, IL Formisano C., Mignola E., Rigano D., Senatore F., Bellone G. et al.: Flavour Frag. J. 22, 289 (2007). 14. Miyazawa M., Yamafuji C., Kurose K., Ishikawa Y.: Flavour Frag. J. 18, 15 (2003). 15. Miyazawa M., Marumoto S., Kobayashi T., Yoshida S., Utsumi Y.: Rec. Nat. Prod. 5, 3 (2011). 10. Pino J.A., Mesa J., Munoz Y., Marti M.P., Marbot R.: J. Agric. Food Chem. 53, 2213 (2005). Received: