Journal of Essential Oil Bearing

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Chemical Characterization of Essential Oils of glaucus ssp. Coronopifolius (Maire) Alexander and Ridolfia segetum (L.) Moris Growing in

Khadija Basaid , Bouchra Chebli , Rachid Bouharroud , James Nicholas Furze , Alessandra Lopes de Oliveira & El Hassan Mayad

To cite this article: Khadija Basaid , Bouchra Chebli , Rachid Bouharroud , James Nicholas Furze , Alessandra Lopes de Oliveira & El Hassan Mayad (2020) Chemical Characterization of Essential Oils of Senecio￿glaucus ssp. Coronopifolius (Maire) Alexander and Ridolfia￿segetum (L.) Moris Growing in Morocco, Journal of Essential Oil Bearing Plants, 23:5, 918-930, DOI: 10.1080/0972060X.2020.1818634 To link to this article: https://doi.org/10.1080/0972060X.2020.1818634

Published online: 07 Dec 2020.

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Full Terms & Conditions of access and use can be found at https://www.tandfonline.com/action/journalInformation?journalCode=teop20 TEOP 23 (5) 2020 pp 918 - 930 918 ISSN Print: 0972-060X ISSN Online: 0976-5026

Chemical Characterization of Essential Oils of Senecio glaucus ssp. coronopifolius (Maire) Alexander and Ridolfia segetum (L.) Moris Growing in Morocco

Khadija Basaid 1, Bouchra Chebli 1*, Rachid Bouharroud 2, James Nicholas Furze 3,5,6, Alessandra Lopes de Oliveira 4, El Hassan Mayad 1,3

1 Laboratory of Mechanic Process Energy and Environment, Biotechnology and environment engineering team, National School of Applied Sciences, Ibn Zohr University, Agadir, Morocco 2 Research Unit of Integrated Crop Production, Centre Regional de la Recherche Agronomique d’ Agadir (INRA), Morocco 3 Laboratory of Biotechnology and Valorization of Natural Resources, Faculty of Sciences- Agadir, Ibn Zohr University, BP 8106, 80000 Agadir, Morocco 4 Laboratory of High-Pressure Technology and Natural Products, Department of Food Engineering, University of São Paulo (USP), Av. Duque de Caxias Norte, 225, Pirassununga, SP 13635-900, Brazil 5 Control and Systems Engineering Department, University of Technology, Baghdad, Alsinaah Street, P.O. Box: 19006, Postal Code: 10066, Baghdad, 6 Royal Geographical Society (with the Institute of British Geographers), 1 Kensington Gore, SW7 2AR, London, UK Received 02 April 2020; accepted in revised form 29 August 2020

Abstract: Chemical profiles of essential oils hydrodistilled from whole plants of Senecio glaucus ssp. coronopifolius (Maire) Alexander and aerial parts of Ridolfia segetum (L.) Moris growing in Morocco were determined using gas chromatography/mass spectrometry (GC/MS). The analysis identified 84 compounds in the oil isolated from S. glaucus ssp. coronopifolius and 61 constituents in R. segetum oil. Both oils had a predominance of monoterpene hydrocarbons. α-Pinene (26.2 %) was the main compound of S. glaucus ssp. coronopifolius oil, followed by myrcene (11.4 %) and p-cymene (9.9 %). Novelly (Z)-β-ocimene (19.7 %) dominated R. segetum oil, followed by β-phellandrene (9.6 %) and β-pinene (8.6 %). This is the first report of (Z)-β- ocimene-rich essential oil from R. segetum and α-pinene-rich essential oil from S. glaucus ssp. coronopifolius, revealing new chemical profiles for both species. The oils have potential in agroecological and pharmaceutical formulations, serving both economic and medicinal applications.

Key words: essential oil, Senecio glaucus ssp. coronopifolius, Ridolfia segetum, Morocco, agroecological.

Introduction glaucus L. is an annual herb that belongs to this Senecio L. ( Cass.) is the largest genus. It has two subspecies: Senecio glaucus genus of the family. It includes over ssp. glaucus and Senecio glaucus ssp. corono- 1500 species distributed worldwide. Amongst pifolius (Maire) Alexander. The latter has a wide these, 22 species are found in Morocco 1. Senecio distribution, ranging accross the Canary Islands,

*Corresponding author (Bouchra Chebli) E-mail: < [email protected] > © 2020, Har Krishan Bhalla & Sons Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 919 through North Africa in Southern Morocco, α-phellandrene (39.4-69.5 %) and terpinolene (6- and Egypt to South-West Asia 2. Although there 27.6 %) 16, and p-cymene (8.8-83.6 %), α-phellan- are no formal reports on the traditional uses of drene (32.0-33.8 %) and terpinolene (18.0-21.4 ssp. Coronopifolius, Senecio sp. have found use %) 23. Further, the Tunisian chemotype of the EO in folk medicine as anti-inflammatory, antiemetic, contained α-phellandrene (34.7-47.8 %) and vasodilatator 3, hypoglycemic drug 4 and for treat- terpinolene (9-23.7 %) 19. The second type of EO ment of coughs, fever, colds and hepatic disorders of the species contains phenylpropanoids as major 5, eczema, wounds, bronchitis and asthma 6. compounds, usually myristicin and dillapiol. This Biological functions of Senecio sp. include its type was found to contain in Morocco α- antioxidant 7, anti-inflammatory 7, antibacterial 8, phellandrene (55 %) and myristicin (33 %) 13; in antifungal 9, insect-repellent 5, antitubercular, Tunisia dillapiole (29.5-85.4 %) and myristicin molluscicidal 10 and cytotoxic 7 activities. Chemical (13.1-31.5 %) 20; in myristicin (70.8 %) 15, in composition of S. glaucus ssp. coronopifolius dillapiol (0.1-45.7 %) and α-phellandrene essential oil (EO) indigenous to Egypt, predomi- (5.2-56.9 %) 16 and dillapiole (5.1-39.6 %) and β- nantly has myrcene (24.0 %) and dehydrofukinone pinene (10.3-20.9 %) 23. (21.0 %) compounds 11. However, chemical Chemical composition of R. segetum EO varies studies have not been carried out on the species according to its origin in flowers, stems, leaves or collected from other geographic locations which roots 15,16,23, extraction method 15,22, and geo- cover its wider distribution. graphical origin 16,19,20,23. The latter indicates the Ridolfia segetum (L.) Moris belongs to the presence of a chemical polymorphism within Apiaceae family, and is the only species of the different populations of the species. We monotypic genus Ridolfia Moris 12. It is a steno- investigated the chemical composition of EOs Mediterranean species, distributed in Morocco 13, obtained from S. glaucus ssp. coronopifolius and Tunisia 14, Sardinia 15, 12, Spain 16 and the R. segetum (L.) Moris occurring in Morocco. This Canaries 17. R. segetum is used in traditional medi- study represents the first characterisation of R. cine to prevent coughing, constipation, respiratory segetum aerial parts EO and reports EO tract infections and for treatment of gastric acidity composition of S. glaucus ssp. coronopifolius 12. It has carminative, antispasmodic, antihaemo- from a previously unstudied site. rrhoidal and emmenagogue properties. Additio- nally, dried fruits of the are used as a eupep- Materials and methods tic digestive in atony and digestive difficulty 18. Plant materials The frequent use of the species in traditional One kg of S. glaucus ssp. coronopifolius (stems, medicine resulted in the identification of several leaves, flowers and roots) and one kg of aerial biological properties, such as antioxidant 12,14, anti- parts of R. segetum (flowers, stems, fruits) were bacterial 14,19,20, anti-inflammatory 12, anticancer collected during flowering and ripening stages, 21 and HIV-1-inhibition 22. Previous studies have from the Massa locality in the south-west of reported the chemical profile of R. segetum EO Morocco (30° 0' 13.6" N 9° 38' 19.6" W), during from different geographic locations, allowing the April, May and June of 2017. Identification of plant distinction of two types of oils. The first type is material was verified by Dr. James N. Furze dominated by monoterpene hydrocarbons, usually (Royal Geographical Society, London, UK). α-phellandrene, terpinolene, and p-cymene. EO Voucher specimens of S. glaucus ssp. corono- of R. segetum from Portugal was found to include pifolius: no. SG-17 and R. segetum no. RS-17) α-phellandrene (53-63.3 %) and terpinolene (8.6- were deposited in the herbarium of the Laboratory 11.9 %) 12, and α-phellandrene (63.3 %) and of Biotechnology, National School of Applied terpinolene (11.9 %) 21; whilst that of Italy con- Sciences, Ibn Zohr University, Agadir, Morocco. tained α-phellandrene (24.7 %) and terpinolene (19.9 %) 15, and α-phellandrene (53.3 %) and Essential oil extraction terpinolene (20.4 %) 22; whilst that of the EO of Plant matter was air dried in the shade. To obtain the species in Spain was profiled and contained the EOs, 250 g of each plants material was soaked Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 920 in 1 L of distilled water in 2 L flasks. Each volume Results and discussion was subjected to hydrodistillation using a The oil yield was expressed as a percentage Clevenger-type apparatus, in a thermo regulated (g/100 g of dried plant material). The average heating mantle for 4 h at 50°C. Hydrodistillation yields of EOs extracted from S. glaucus ssp. was repeated four times for each plant sample coronopifolius and R. segetum were found to and the average yield was calculated. The mixture be 0.2 g and 0.125 g from 250 g of each plant, of distillated oils and water was collected, the upper and the percentage of EO yields were 0.08 and partition of EOs were separated from the lower 0.05 % respectively. De Pooter et al.11 reported water phase, weighed and stored in tightly closed a slightly higher yield of 0.25 % for Egyptian S. dark vials at 4°C for further analysis. glaucus ssp. coronopifolius oil obtained by steam distillation. Whereas, Ramadan et al.7 reported Chemical composition of essential oils yields of 0.5 and 0.25 % for capetula and shoots The chemical composition of EOs was identified oils of Egyptian S. glaucus L. For R. segetum, and quantified by gas chromatography coupled Cabral et al.12 reported higher yields of 1.6 and with mass spectrometry (GC/MS) 24. The GC was 1.9 % in flowering and ripening stages respectively carried out using a QP 2010 Plus gas chromato- of the species growing in Portugal, by hydro- graphy apparatus (Shimadzu, Tokyo, JP) equipped distillation. Conversely, yields of 0.4 and 0.94 % with an automatic injector (AOC-5000, Belle- were obtained using steam distillation and super- fonte, USA), and an RTX-5MS capillary column critical CO2 respectively on R. segetum of Italy (30 m × 0.25 mm ID × df 0.25 μm, 5 % diphenyl/ 15,22. Variation in yield appears to be related to the 95 % dimethyl polysiloxane) (Restek, Belle-fonte, extraction method and parameters. The influence USA). Helium was used as a carrier gas at a of the duration of distillation on EO yield has been pressure of 100, 2 KPa and a linear velocity of reported 20, in addition to the vegetative stage of 46.3 cm/s. Fragmentation was effected by elec- plants 27, the season of harvest 28, geographic loca- tronic impact. 1 μL of EO diluted in dichloro- tion 29, and growth conditions of the plants 30. methane (Merck, Darm-stadt, GE) at a concen- GC/MS analysis of S. glaucus ssp. corono- tration of 1 % (10 mg/ml) was injected using 1:10 pifolius oil enabled the identification of eighty- split ratio. The injector temperature was 250°C four compounds corresponding to 94.3 % of the and the temperature at the interface between the total constituents. 86.3 % of the total identified column and the ion source was set to 250°C. The compounds were terpenic, monoterpene hydro- oven temperature was regulated to obtain a linear carbons were the most prominent (59.4 %), ramp from 60 to 246°C (3°C/min). The acquisition followed by oxygenated sesquiterpenes (11.1 %), mass range was 50 to 470 m/z, at a speed of 909 sesquiterpene hydrocarbons (8.9 %), and oxy- u/s, using electron ionization (EI) with electron genated monoterpenes (6.9%) (Table 1). The energy of 70 eV. predominance of monoterpene hydrocarbons has The EOs components were identified by been reported for EOs of several species of the matching their mass spectra value to NIST11 and genus Senecio 7. Alternatively, few species of this NIST11s database libraries, using the GC-MS genus were dominated by oxygenated compounds, Solutions V2.5 software. The Van der Dool and as was found in S. chrysanthemoides, S. verna- Kratz Index was calculated as an additional tool lis and S. aegyptius var. discoideus 31, 32, 33. for compounds identification 25. Quantification of α-Pinene was identified as the main compound the oils components was performed by calculation in ssp. coronopifolius EO at 26.2 %, followed of their retention indices (RI exp) using a mixture by myrcene (11.4 %) p-cymene (9.9 %), β-pinene γ of hydrocarbons (C10H22-C40H82) (Fluka, St. (7.7 %), -muurolene (4.0 %), deoxynivalenol (3.1 Louis, USA) as standard. Components were %) and α-phellandrene (2.7 %) (Table 1). Analysis further confirmed by comparison of their retention of oil extracted from aerial parts of S. glaucus indices with those reported in the literature (RI ssp. coronopifolius material collected in Egypt lit) 26. 11 showed myrcene (24.0 %), dehydrofukinone Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 921 Table 1. Chemical composition of the essential oil isolated from Senecio glaucus ssp. coronopifolius

No. Compound RIexp RIlit Relative content (%)

1 1-Isopropyl-4-methylenebicyclo [3.1.0]hex-2-ene 879 - 0.2 2 Cyclopropane, 1-methyl-2-pentyl- 880 - 1.0 3 α-Thujene 922 924 0.1 4 α-Pinene 930 932 26.2 5 Camphene 943 946 0.1 6 β-Pinene 973 974 7.7 7 Myrcene 985 988 11.4 8 Decane 1000 1000 0.1 9 α-Phellandrene 1001 1002 2.7 10 α-Terpinene 1011 1014 0.2 11 p-Cymene 1022 1020 9.9 12 γ-Terpinene 1050 1054 0.5 13 Ethanol, 1-(1-cyclohexenyl)- 1053 - 0.1 14 Terpinolene 1082 1086 0.3 15 Perillene 1101 1102 0.5 16 1-Octen-3-yl-acetate 1109 - 1.0 17 α-Campholenal 1120 1122 0.8 18 cis-Verbenol 1136 1137 0.4 19 2-Caren-10-al 1136 - 0.1 20 Pinocarvone 1157 1160 0.2 21 Naphthalene, 1,2,3,4,4a,5,8,8a-octahydro- 1157 - 0.6 4a-methyl-, trans- 22 Terpinen-4-ol 1173 1174 1.3 23 Phellandral 1175 - 0.3 24 Isopinocarveol 1180 - 0.4 25 α-Terpineol 1183 1186 0.4 26 p-Mentha-1(7),8-dien-2-ol 1188 1187 0.1 27 (1R)-(-)-Myrtenal 1194 1195 0.5 28 1-Methylverbenol 1209 - 0.1 29 4(10)-Thujen-3-ol, acetate 1224 - 0.1 30 p-Mentha-6,8-dien-2-ol, cis- 1226 1227 0.2 31 Cumaldehyde 1230 - 0.3 32 Isoaromadendrene epoxide 1281 - 0.5 33 p-Cymen-7-ol 1284 1289 0.1 34 2-Undecanone, 6,10-dimethyl- 1291 1293 0.1 35 Ledene oxide-(II) 1293 - 1.3 36 Carvacrol 1295 1298 0.4 37 2-(1,4,4-Trimethyl-cyclohex-2-enyl)-ethanol 1313 - 0.2 38 3a-Methyl-3a,4,5,6,7,8-hexahydro-2(3H)-azulenone 1357 - 0.1 39 (-)-β-Bourbonene 1386 1387 0.2 40 Alloaromadendrene 1386 - 0.2 41 (-)-cis-β-Elemene 1387 1389 0.4 42 (+)-Sativene 1390 1390 0.1 Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 922 table 1. (continued).

No. Compound RIexp RIlit Relative content (%)

43 Caryophyllene 1404 1408 0.6 44 α-Guaiene 1434 1437 0.4 45 Aromadendrene oxide-(2) 1435 1439 0.6 46 Di-epi-α-Cedrene 1438 1440 0.1 47 Seychellene 1441 1444 0.1 48 α-Humulene 1450 1452 0.1 49 Cycloisolongifolene, 8,9-dehydro-9-formyl- 1457 - 0.1 50 Aromadendrene oxide-(1) 1457 1458 0.1 51 Cadina-3,9-diene 1458 1461 0.5 52 Cyclohexene, 3-acetoxy-4-(1-hydroxy- 1471 - 0.8 1-methylethyl)-1-methyl- 53 Bicyclo[10.1.0]tridec-1- 1472 - 0.1 54 γ-Muurolene 1477 1478 4.0 55 4,5-di-epi-Aristolochene 1488 1487 0.3 56 β-Vatirenene 1489 - 0.3 57 2-Isopropenyl-4a,8-dimethyl-1,2,3,4,4a, 1502 - 0.5 5,6,7-octahydronaphthalene 58 Curcumene 1512 1514 0.9 59 Cubenol 1515 1514 0.2 60 Eremophila-1(10),11-diene 1557 1558 1.8 61 Spathulenol 1576 1577 1.0 62 Caryophyllene oxide 1580 1582 0.5 63 Tricyclo[5.2.2.0(1,6)]undecan-3-ol, 2- 1599 - 1.2 methylene-6,8,8-trimethyl- 64 Longipinocarveol, trans- 1599 - 0.1 65 Tau-muurolol 1641 1644 0.2 66 Guaia-1(10),11-diene 1674 1676 0.1 67 6-Isopropenyl-4,8a-dimethyl-1,2,3,5, 1690 - 0.1 6,7,8,8a-octahydronaphthalen-2-ol 68 2-(4a,8-Dimethyl-1,2,3,4,4a,5,6,7-octahydro 1745 - 0.2 -naphthalen-2-yl)-prop-2-en-1-ol 69 Hexahydrofarnesyl acetone 1754 - 0.7 70 Eudesma-5,11(13)-dien-8,12-olide 1801 - 0.2 71 6.β.Bicyclo[4.3.0]nonane, 5.β.-iodomethyl- 1.β.-isopropenyl-4α,5α-dimethyl- 1831 - 0.1 72 Podocarp-13-en-12-ol 1861 - 0.1 73 Anthracene, 1,2,3,4,5,6,7,8-octahydro-9,10-dimethyl- 1879 - 1.4 74 β-Springene 1922 - 0.1 75 Phytol 1940 1942 0.2 76 Phytol, acetate 1941 1942 0.1 77 Geranyl linalool 2026 2026 0.2 78 2-Butenoic acid, 2-methyl-, 1a,2,4,4a,5,9- 2109 - 0.9 hexahydro-4,4a,6-trimethyl-3H-oxireno[8,8a] naphtho[2,3-b]furan-5-yl ester Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 923 table 1. (continued).

No. Compound RIexp RIlit Relative content (%)

79 2-Butenoic acid, 2-methyl-, 4,4a,5,6,7,9- 2219 - 0.1 hexahydro-3,4a,5-trimethylnaphtho[2,3-b] furan-4-yl ester, [4S-[4.α.(Z),4a.α.,5.α.]]- 80 Kauran-18-al, 17-(acetyloxy)-, (4.β.)- 2338 - 0.1 81 Deoxynivalenol 2385 - 3.1 82 Heneicosane 2395 2400 0.2 83 4-Piperidyl cyclopentylphenylglycolate 2498 - 0.2 84 β-d-Mannofuranoside, farnesyl 3014 - 0.3 Monoterpene hydrocarbons [No. 1, 3-12, 14] 59.4 Oxygenated monoterpenes [No. 15-20, 22-28, 31, 33, 36] 6.9 Sesquiterpene hydrocarbons [No. 39-44, 46-48, 51, 54-58, 66, 71] 8.9 Oxygenated sesquiterpenes [No. 32, 35, 45, 50, 59-65, 67, 68, 70, 81] 11.1 Others [No. 2, 13, 21, 29, 30, 34, 37, 38, 49, 52, 53, 69, 72-80, 82-84] 8.0 Total 94.3

= RIexp Retention indices determined on a 30 m Rtx-5 capillary column using a mixture of hydrocarbons = 26 (C10H22-C40H82).; RIlit Retention indices reported in the literature (21.0 %) and p-cymene (9.9 %) to be the major oils of different origin. The major compounds of compounds of the oil. Myrcene and p-cymene capetula oil were m-mentha-1(7),8-diene (31.4 were found as major compounds of S. glaucus %), cis-m-mentha-2,8-diene (22.9 %), dehydro- ssp. coronopifolius oil in the current study at 11.4 fukinone (17.2 %) and α-terpinolene (3.9 %). and 9.9 % respectively, whereas dehydrofukinone Whereas, the shoots oil contained m-mentha- wasn’t detected. Comparatively, α-pinene, the 1(7),8-diene (25.6 %), cis-m-mentha-2,8-diene main compound detected in oil of the current study, (8.2 %), dehydrofukinone (19.9 %), α-fenchene though was found in Egyptian oil in lower (5.6 %), 1,3,8-p-menthatriene (5.3 %) and β- proportions (2.6 % vs 26.2 %). Moreover, the ocimene (3.4 %) as the main constituents. Plant current study enabled the identification of a higher part plays a crucial role in the accumulation pattern number of compounds in the oil (84 compounds), and chemical composition of EOs 34. Production whereas, in Egyptian oil, De Pooter et al.11 of plant terpenes is mainly controlled by trans- identified 31 compounds. Thus, a new chemical criptional regulation of terpene synthase genes, profile of S. glaucus ssp. coronopifolius EO is which have organ-specific expression patterns. reported herein. Variability in chemical composition The spatial regulation of terpene synthase gene of the species EO may be related to the difference expression qualitatively contributes to terpene in plant parts used for extraction of the oil. In composition 35. Thus, the occurrence of certain experimentation of De Pooter et al.11, the EO was molecules in higher amounts such as α-pinene extracted from aerial parts, whereas in the current could be attributed to their presence in roots oil. case, oil was extracted from whole plants, including Other factors potentially responsible for the chemi- aerial parts and roots. A more recent study by cal profile obtained of S. glaucus ssp. corono- Ramadan et al.7 supported that variation in oil pifolius oil are a complex of genetic and/or composition is in accordance with the plant organ. environmental interactions 36. It reported the composition of capetula and shoots There is a similarity between S. glaucus ssp. EOs of S. glaucus L. from Egypt, and indicated coronopifolius and other Senecio species in qualitative and quantitative variations between the terms of the main constituents of EOs. α-pinene Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 924 (26.2 %) and p-cymene (9.9 %), which are major parts. The profile of the current work indicates a compounds of the oil have been reported as the distinct chemotype of R. segetum, which may be main constituents in other species at different specific to Moroccan ecotypes. amounts. S. farfarifolius Boiss. Et Kotschy EO Variation in chemical composition of R. segetum from Turkey was reported to contain á-pinene at EO according to geographical origin reveals a higher percentage (48.3 %) 37, and EO of S. differences among countries, and also between ambavilla (Bory) Pers. from contained different collection sites within each country. In α-pinene at a lesser amount (14 %) 38, while p- Tunisia, flower oil of the species collected from cymene was found in EO of S. nutans Sch. Bip, the region of Kroussia contained α-phellandrene grown in the Luara region of Peru, Southern (34.7 %) and terpinolene (23.7 %) as the main America at 8.8 % 39. constituents 19, whereas the EO of flowers Analysis of R. segetum oil led to identification collected from the region of Ouled Alouène was of sixty-one components, accounting for 84.7 % reported to be dominated by dillapiole (29.5-85.4 of the total constituents. 74.4 % of the total identi- %) and myristicin (13.1-31.5 %) 20. In Spain, EO fied compounds were terpenic. Of the terpenes, of leaves collected from Andalucia province was monoterpene hydrocarbons were the main class dominated by α-phellandrene (61.8-69.5 %) 16, (40.1 %), which comes in agreement with previous whereas the oil of leaves collected from Castilla studies 12,15,16,19,21,22,23, followed by sesquiterpene la Mancha province was dominated by p-cymene hydrocarbons (17.1 %), oxygenated sesqui- (8.8-83.6 %) 23. Stem oil from Andalucia province terpenes (14.7 %) and oxygenated monoterpenes contained mainly α-phellandrene (39.4-62.0 %) (2.5 %). (Z)-β-ocimene (19.7 %) was the main 16, whilst stem oil of the species from Castilla la constituent of the oil followed by β-phellandrene Mancha province was seen to be dominated by (9.6 %), β-pinene (8.6 %), tricyclo [5.1.0.0(3,5)] p-cymene (15.1-79.5 %) 23. Analysis of Moroccan octane-2,6-dione, 1,3,4,5,7,8-hexamethyl-(7.0 %), R. segetum fruit EO showed myristicin (70.2 %) di-epi-α-cedrene (5.4 %), cubenol (2.9 %) and and γ-terpinene (8.2 %) to be major compounds alloaromadendrene (2.5 %) (Table 2). 40. Although, fruits were used in the current study Reports on EO derived from aerial parts of R. for extraction of EO, γ-terpinene was detected segetum show α-phellandrene, terpinolene, β- only in traces (0.1 %) whereas myristicin was phellandrene, p-cymene, β-pinene and (Z)-β- not detected, probably owing to a difference in ocimene to be characteristic constituents of leaves, regions of plant collection. El Karkouri et al. 40 flowers and stem oils. In concordance, the present documented R. segetum from the north-west of study identified (Z)-β-ocimene, β-phellandrene Morocco, whereas the present study was carried and β-pinene as the most abundant constituents, out on plant material from the south-west. A whilst α-phellandrene and terpinolene were gradient from a cooler, humid climate to hotter, detected at trace (0.1 %) levels, presence at such drier climes exists from northern to southern a level indicates they serve an activator function regions. Consequently, local ecological factors act amongst the compound family. Further, consti- on the biochemical pathways of terpene production tuents found in fruit oil (phenylpropanoids 41, contributing to the appearance of different myristicin, dillapiole, and piperitenone oxide) chemical profiles 42. 13,15,16,23,40 were not detected in R. segetum oil of The choice of organ used for extraction also the present study, indicating a region specific influences composition of R. segetum oil. In Spain, chemotype. α-Phellandrene was reported as the the oil extracted from leaves and stems was main constituent of aerial parts of R. segetum oil characterized by the compounds p-cymene (8.8- in Italy, Portugal, Spain and Tunisia at 53.3 %, 83.6 %) and (Z)-β-ocimene (traces-38.5 %). The 63.3 %, (39.4-69.5 %) and (34.7-47.8 %) respecti- flowers oil was dominated by α-phellandrene vely 12,16,19,22. The present study seminally identi- (32.0-33.8 %) and terpinolene (18.0-21.4 %), and fies and reports (Z)-β-ocimene as the number one the fruits oil contained mainly dillapiole (5.1-39.6 compound in R. segetum oil derived from aerial %) and β-pinene (10.3-20.9 %) 23. Likewise, Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 925 Table 2. Chemical composition of the essential oil isolated from the aerial parts of Ridolfia segetum (L.) Moris

No. Compound RIexp RIlit Relative content (%)

1 α-Thujene 922 924 0.3 2 Camphene 943 946 0.5 3 Carane, 4,5-epoxy-, trans 948 - 0.1 4 β-Pinene 973 974 8.6 5 α-Phellandrene 1000 1002 0.1 6 o-Cymene 1019 1022 1.0 7 β-Phellandrene 1023 1025 9.6 8 (Z)-β-Ocimene 1031 1032 19.7 9 (Z)-β-Ocimene 1038 1032 0.1 10 γ-Terpinene 1050 1054 0.1 11 Terpinolene 1082 1086 0.1 12 Camphenone, 6- 1094 1095 0.1 13 α-Campholenal 1125 1122 0.3 14 Pinocarveol 1131 1135 0.3 15 cis-Verbenol 1136 1137 0.4 16 Pinocarvone 1157 1160 0.1 17 Ethanone, 1-(1,4-dimethyl-3-cyclohexen-1-yl)- 1161 - 0.1 18 Terpinen-4-ol 1173 1174 0.5 19 Naphthalene, 1,2,3,4,4a,5,8,8a-octahydro- 1177 1178 0.5 4a-methyl-, trans- 20 α-Terpineol 1183 1186 0.2 21 (-)-Myrtenol 1191 1194 0.3 22 cis-Carveol 1226 1226 0.1 23 Isoaromadendrene epoxide 1281 - 0.3 24 Ledene oxide-(II) 1293 - 0.1 25 1H-Indene, 2,3,3a,4,7,7a-hexahydro-2,2,4, 1367 - 0.1 4,7,7-hexamethyl-, trans- 26 Copaene 1371 1374 0.3 27 trans-2-Dodecen-1-ol, pentafluoropropionate 1383 - 0.1 28 Longipinene 1400 1400 1.2 29 (+)-Calarene 1403 - 1.0 30 Di-epi-α-Cedrene 1407 1410 5.4 31 Cadala-1(10),3,8-triene 1423 - 0.5 32 α-Guaiene 1436 1437 0.6 33 2-Butanone, 4-(2,6,6-trimethyl-2- 1445 - 0.1 cyclohexen-1-ylidene)- 34 Geranyl acetone 1451 1453 0.2 35 Bicyclo[5.3.0]decane, 2-methylene-5- 1456 - 0.5 (1-methylvinyl)-8-methyl- 36 Alloaromadendrene 1456 1458 2.5 37 cis-(-)-2,4a,5,6,9a-Hexahydro-3,5,5,9- 1471 - 1.1 tetramethyl (1H)benzocycloheptene Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 926 table 1. (continued).

No. Compound RIexp RIlit Relative content (%)

38 Cycloheptane, 4-methylene-1-methyl-2- 1475 - 0.1 (2-methyl-1-propen-1-yl)-1-vinyl- 39 γ-Muurolene 1475 1478 0.6 40 3-Methyl-2-butenoic acid, 2-adamantyl ester 1492 - 0.4 41 2-(3-Isopropyl-4-methyl-pent-3-en-1-ynyl)- 1496 - 1.4 2-methyl-cyclobutanone 42 β-Himachalene 1500 1500 0.1 43 Chamigrene 1503 1503 2.0 44 α-Farnesene 1504 1505 0.4 45 Farnesene epoxide, E- 1507 1505 0.9 46 γ-Cadinene 1511 1513 0.7 47 Cubenol 1514 1514 2.9 48 Tricyclo[5.1.0.0(3,5)]octane-2,6-dione, 1522 - 7.0 1,3,4,5,7,8-hexamethyl- 49 Cyclohexanemethanol, 4-ethenyl-α, α, 4-trimethyl 1522 - 1.7 -3-(1-methylethenyl)-, [1R-(1α, 3α, 4 β)]- 50 Hedycaryol 1545 1546 0.8 51 Spathulenol 1576 1577 0.2 52 Carotol 1593 1594 0.8 53 Rosifoliol 1598 1600 0.8 54 γ-Eudesmol 1626 1630 2.1 55 Alloaromadendrene oxide-(1) 1635 1639 0.5 56 α-Bisabolol 1685 1685 1.9 57 6-Isopropenyl-4,8a-dimethyl-1,2,3,5,6,7,8, 1690 - 0.7 8a-octahydro-naphthalen-2-ol 58 5(1H)-Azulenone, 2,4,6,7,8,8a-hexahydro- 1694 - 0.6 3,8-dimethyl-4-(1-methylethylidene)-, (8S-cis)- 59 Geranyl-α-terpinene 1962 - 0.7 60 3-Ethyl-3-hydroxyandrostan-17-one 2251 - 0.1 61 Hexacosa-2,25-dione 2878 - 0.2 Monoterpene hydrocarbons [No. 1, 2, 4-11] 40.1 Oxygenated monoterpenes [No. 3, 12-18, 20-22] 2.5 Sesquiterpene hydrocarbons [No. 25, 26, 28-32, 35-39, 42-44, 46] 17.1 Oxygenated sesquiterpenes [No. 23, 24, 40, 45, 47, 49-58] 14.7 Others [No. 19, 27, 33, 34, 41, 48, 59-61] 10.3 Total 84.7

= RIE Retention indices determined on a 30m Rtx-5capillary column using a mixture of hydrocarbons (C10H22- = 26 C40H82); RIlit Retention indices reported in the literature . Marongiu et al.15 showed that flowers and stems differences may be attributed to changes in oils were dominated by α-phellandrene (12.9-19.4 metabolism of EO production in different plant %) and terpinolene (11.6-20.5 %), whereas, fruits organs 43. oil was found rich in myristicin (70.8 %). These Extraction method further influences the Khadija Basaid et al., / TEOP 23 (5) 2020 918 - 930 927 composition of R. segetum oil. Slight differences pounds present in small quantities, which can be in percentages of the main constituents were decisive in inducing biological activity of the oils; noted between oils obtained with hydrodistillation, as the latter is related to the synergetic and/or steam distillation and supercritical CO2 extraction antagonistic interactions between major and minor 15,22. These differences can be related to the constituents 49. Hence, exploring these species for mechanisms used in each extraction method. As their bioactivity against phytopathogens or for EO compounds are not fully soluble in water, production of active compounds is required as a hydrodistillation causes compounds to migrate continuum of this study. from the inside of plant leaves up to their surface, followed by their subsequent evaporation. Hence, Conclusions the vegetable matrix releases only low molecular The present study investigated the chemical weight compounds. Whereas, supercritical CO2 composition of EOs extracted from S. glaucus enables extraction of high molecular weight com- ssp. coronopifolius and R. segetum growing in pounds from the plant particles 15. Duration/time- Morocco. Essential oil of S. glaucus ssp. corono- periods of extraction affect chemical composition. pifolius was seen to be rich in monoterpene Extraction of lighter compounds such as mono- hydrocarbons, with á-pinene being the major terpene hydrocarbons contrasts with oxygenated constituent. R. segetum oil was also rich in mono- sesquiterpenes and heavier hydrocarbons in terpene hydrocarbons, though (Z)-β-ocimene was extraction time 15. Major compounds of R. sege- the main constituent of the EO. Qualitative and tum flower oil from Tunisia vary according to the quantitative differences in composition were time / length of the hydrodistillation 20. Similarily, noticed in comparison with oils of the same species prolonged steam distillation of Italian R. segetum from different countries, suggesting the existence gave a chemically more diverse EO profile 44. of chemical diversity within both species. Further Chemical profiles obtained of S. glaucus ssp. analysis such as cluster analysis and dendogram coronopifolius and R. segetum EOs may also structures are suggested to confirm that the be an outcome of the analysis method. Variation populations of S. glaucus ssp. coronopifolius and in the choice of the analytic method or in experi- R. segetum growing in Morocco are indeed new mental parameters of the method (such as the chemotypes. Comparisons of chemical dissimilari- type of column employed) results in dissimilarities ties and homologues, as well as calculations of of the molecules detected. This comes in the matrices of genetic background of S. glaucus agreement with findings of Fan et al.45, who ssp. coronopifolius and R. segetum enable reported a difference in the coverage of EO com- further investigations which facilitate under- position using different columns. standing of species inter-relations and functions Characterizing the composition of plant species 50. Comprehensive knowledge of the chemical metabolites will enable decision making on composition of EOs provides crucial background domestication and breeding programs ultimately to further investigations, regarding the outcome according to the desired essential oil profile. of the oils with pathogenic life forms of fungi, Essential oils display different levels of biological nematodes, insects and bacteria. activities according to their chemical composition 42. Chemical profiles of S. glaucus ssp. corono- Conflict of interests pifolius and R. segetum EOs obtained herein were No potential conflict of interest was reported characterized by the presence of many bioactive by the authors. compounds, which might have several applications in crop protection. 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