Chemical Characterization of Marrubium vulgare Volatiles from Serbia

Lato  Pezo

Marrubium vulgare is a cosmopolitan medicinal plant from the Lamiaceae family, which produces structurally highly diverse groups of secondary metabolites. A total of 160 compounds were determined in the volatiles from Serbia during two investigated years (2019 and 2020). The main components were E-caryophyllene, followed by germacrene D, α-humulene and α-copaene. All these compounds are from sesquiterpene hydrocarbons class which was dominant in both investigated years. This variation in volatiles composition could be a consequence of weather conditions, as in the case of other aromatic plants. According to the unrooted cluster tree with 37 samples of Marrubium sp. volatiles from literature and average values from this study, it could be said that there are several chemotypes: E-caryophyllene, β-bisabolene, α-pinene, β-farnesene, E-caryophyllene + caryophyllene oxide chemotype, and diverse (unclassified) chemotypes. However, occurring polymorphism could be consequence of adaptation ...

plants Article Chemical Characterization of Marrubium vulgare Volatiles from Serbia Milica Aćimović 1, * , Stefan Ivanović 2 , Katarina Simić 2 , Lato Pezo 3 , Tijana Zeremski 1 , Jelena Ovuka 1 and Vladimir Sikora 1 1 2 3 * Citation: Aćimović, M.; Ivanović, S.; Simić, K.; Pezo, L.; Zeremski, T.; Ovuka, J.; Sikora, V. Chemical Characterization of Marrubium vulgare Volatiles from Serbia. Plants 2021, 10, 600. https://doi.org/10.3390/ plants10030600 Institute of Field and Vegetable Crops Novi Sad, Maksima Gorkog 30, 21000 Novi Sad, Serbia; tijana.zeremski@ifvcns.ns.ac.rs (T.Z.); jelena.ovuka@ifvcns.ns.ac.rs (J.O.); vladimir.sikora@ifvcns.ns.ac.rs (V.S.) Institute of Chemistry, Technology and Metallurgy, University of Belgrade, 11000 Belgrade, Serbia; stefan.ivanovic@ihtm.bg.ac.rs (S.I.); katarina.simic@ihtm.bg.ac.rs (K.S.) Institute of General and Physical Chemistry, University of Belgrade, 11000 Belgrade, Serbia; latopezo@yahoo.co.uk Correspondence: milica.acimovic@ifvcns.ns.ac.rs Abstract: Marrubium vulgare is a cosmopolitan medicinal plant from the Lamiaceae family, which produces structurally highly diverse groups of secondary metabolites. A total of 160 compounds were determined in the volatiles from Serbia during two investigated years (2019 and 2020). The main components were E-caryophyllene, followed by germacrene D, α-humulene and α-copaene. All these compounds are from sesquiterpene hydrocarbons class which was dominant in both investigated years. This variation in volatiles composition could be a consequence of weather conditions, as in the case of other aromatic plants. According to the unrooted cluster tree with 37 samples of Marrubium sp. volatiles from literature and average values from this study, it could be said that there are several chemotypes: E-caryophyllene, β-bisabolene, α-pinene, β-farnesene, E-caryophyllene + caryophyllene oxide chemotype, and diverse (unclassified) chemotypes. However, occurring polymorphism could be consequence of adaptation to grow in different environment, especially ecological conditions such as humidity, temperature and altitude, as well as hybridization strongly affected the chemotypes. In addition, this paper aimed to obtain validated models for prediction of retention indices (RIs) of compounds isolated from M. vulgare volatiles. A total of 160 experimentally obtained RIs of volatile compounds was used to build the prediction models. The coefficients of determination were 0.956 and 0.964, demonstrating that these models could be used for predicting RIs, due to low prediction error and high r2 . Academic Editor: Jésus Palá-Pául Keywords: horehound; GC–MS; retention indices; QSRR; boosted trees regression model Received: 28 February 2021 Accepted: 16 March 2021 Published: 23 March 2021 1. Introduction Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). Marrubium vulgare L., also known as white horehound, is a perennial species from the Lamiaceae family. It is indigenous to the region between the Mediterranean Sea and Central Asia; however, today it is found worldwide, apart from the coldest regions and high altitudes [1]. This plant is highly resistant to drought and due to this it grows well in semiarid areas [2]. Additionally, as it is a moderate salt-tolerant species this medicinal plant could be grown on saline soil [3]. The surface of M. vulgare vegetative and generative organs is densely clothed with glandular and nonglandular trichomes which accumulate secondary metabolites [4]. M. vulgare produces structurally highly diverse groups of secondary metabolites, thus represents a valuable source of bioactive compounds and preparations with health-promoting effects: antioxidant, hepatoprotective, antiproliferative, anti-inflammatory, antidiabetic, and antimicrobial [5]. The use of this herb in traditional medicine is recorded worldwide for ameliorating chronic cough and cold, numerous conditions related to skin, liver, gastric, heart, and immune system [6]. Generally, M. vulgare is poor in essential oil, and the major compounds are diverse [1,7–17]. Plants 2021, 10, 600. https://doi.org/10.3390/plants10030600 https://www.mdpi.com/journal/plants Plants 2021, 10, 600 2 of 17 This proves that there are various chemotypes of M. vulgare. The lack of information in this field is pointed out by Yabrir [1,18]. The studies about genus Marrubium are mainly focused on taxonomical, morphological, and genetic diversity [4,19–26]. The main aim of this investigation was to determine volatiles composition of M. vulgare grown in Serbia during two years and to compare its chemical composition with literature data not only of M. vulgare but with other species from this genus as well (M. anisodon, M. aschersonii, M. astracanicum, M. crassidens, M. deserti, M. duabense, M. parviflorum, M. peregrinum, M. persicum, M. propinquum, M. velutinum). Another goal was to establish the new quantitative structure retention relationship (QSRR) models for anticipating the retention indices (RIs) of certain compounds in M. vulgare volatiles obtained by GC–MS chromatography utilizing the genetic algorithm (GA) variable selection method and the boosted trees regression. Furthermore, we gather information about the volatile compounds of species from Marrubium genus in order to classify the chemotype of M. vulgare from this study according to unrooted cluster tree. 2. Results The main components in M. vulgare volatiles were E-caryophyllene with 24.6% and 23.0%, followed by germacrene D with 9.6% and 17.0%, α-humulene with 5.2% and 5.3% as well as α-copaene with 3.3% and 6.1% in 2019 and 2020, respectively. All these compounds are from the sesquiterpene hydrocarbons class which was dominant in both years of the investigation, 52.0% in 2019 and 67.8% in 2020. This variation in volatiles composition could be a consequence of weather conditions, as in case of other aromatic plants [27–33]. However, some of the components detected in M. vulgare volatiles during the twoyear research have not been detected yet in this species, while other components have not been detected in other species of this genus. ScienceDirect Elsevier, SpringerLink, PubMed, Scopus, Scifnder, Web of Science, Wiley Online, and Google Scholar databases were reviewed and scientific publications from 1990 until 2020 that deal with chemical composition of volatiles species from genus Marrubium were summarized and shown in Table 1. Table 1. Chemical composition of Marrubium vulgare during two years (2019 and 2020). No Compound/Class Cycle RIpred. 1 2E-Hexenal O Train 2 Furan, 2,5-diethyltetrahydro O Validation 2019 2020 RIa % RIa % 892.915 - - 847 0.2 853.684 - - 897 0.1 Reference M. aschersonii [34], M. deserti [35], M. peregrinum [36], M. vulgare [10,12,15,16,34] M. anisodon [37], M. astracanicum [38], M. crassidens [39], M. deserti [35], M. duabense [40], M. parviflorum [41,42], M. peregrinum [43,44], M. persicum [45], M. propinquum [41], M. velutinum [44], M. vulgare [7,8,10,15] 3 1-Octen-3-ol O Validation 965.818 976 0.2 974 0.6 4 2-Pentyl furan O Train 1059.803 - - 989 0.1 5 3-Octanol O Test 962.233 - - 992 0.1 M. anisodon [37], M. astracanicum [46], M. duabense [40], M. peregrinum [36,44], M. velutinum [44] 6 Linalool OMN Train 1106.041 1102 0.1 1098 0.1 M. aschersonii [34], M. astracanicum [46], M. parviflorum [41,42,47,48], M. peregrinum [36,43,44], M. persicum [45], M. velutinum [44], M. vulgare [8,10,12,17,34,36,47,49] 7 n-Nonanal O Train 1078.484 - - 1102 0.1 M. aschersonii [34], M. deserti [35], M. duabense [40], M. peregrinum [43,44], M. persicum [45], M. velutinum [44], M. vulgare [34] 8 E-Thujone OMN Train 1118.307 - - 1114 0.1 M. peregrinum [43], M. vulgare [8,15] 9 NI-1 - - - - 1132 0.1 - Plants 2021, 10, 600 3 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 10 Geijerene O Train 11 2E-Nonen-1-al O 12 β-Cyclocitral O 13 2019 2020 Reference RIa % RIa % 1192.301 1143 0.1 1139 0.6 Validation 1097.602 - - 1156 0.1 Train 1216.889 - - 1219 0.1 Cogeijerene O Train 1203.235 - - 1283 0.1 14 Pregeijerene O Train 1149.857 1290 0.1 1287 0.2 M. astracanicum [38], M. crassidens [38], M. parviflorum [42,47], M. peregrinum [43] 15 Thymol AR Test 1209.017 1292 0.3 - - M. deserti [52], M. vulgare [7,8,10,15,50] 16 2-Undecanone O Train 1269.228 1295 0.1 1292 Trace M. vulgare [15] - M. duabense [40], M. incanum [50], M. parviflorum [42], M. peregrinum [43], M. vulgare [7,8,10,49,50] 17 Carvacrol AR Validation 1179.072 1302 0.1 - M. incanum [50,51], M. parviflorum [42,47], M. peregrinum [43], M. vulgare [50] M. peregrinum [44], M. velutinum [44], M. vulgare [10] 18 δ-Elemene ST Test 1512.436 - - 1336 0.1 M. anisodon [37], M. astracanicum [38], M. crassidens [38], M. deserti [35,40], M. duabense [40], M. incanum [50,51], M. parviflorum [47], M. peregrinum [44], M. persicum [53], M. thessalum [54], M. velutinum [44], M. vulgare [47,50] 19 α-Cubebene ST Train 1491.202 - - 1348 0.1 M. astracanicum [38], M. crassidens [39], M. deserti [35,40], M. duabense [40], M. parviflorum [47], M. peregrinum [44], M. persicum [45], M. vulgare [8,47] 20 Eugenol AR Train 1372.624 - - 1357 0.4 M. aschersonii [34], M. peregrinum [36,43,44], M. persicum [53], M. velutinum [44], M. vulgare [10,12,34,36,47] 6.1 M. anisodon [37], M. aschersonii [34], M. astracanicum [38], M. crassidens [38], M. deserti [35], M. duabense [40], M. incanum [50,51], M. parviflorum [42,47,48], M. peregrinum [36,43,44], M. persicum [53], M. thessalum [54], M. velutinum [44], M. vulgare [8–13,36,47,50] M. anisodon [37], M. astracanicum [38], M. crassidens [38], M. deserti [35,52], M. incanum [50,51], M. parviflorum [41,42,47,48], M. peregrinum [43,44], M. persicum [45,53], M. thessalum [54], M. velutinum [44], M. vulgare [9,10,13,50] 21 α-Copaene ST Train 1475.878 1377 3.3 1377 22 β-Bourbonene ST Train 1487.976 1385 0.8 1384 1.2 23 NI-2 - - - - 1388 0.1 - 0.2 M. aschersonii [34], M. deserti [35], M. peregrinum [43,44], M. parviflorum [42], M. velutinum [44], M. vulgare [12,13,34,47] M. anisodon [37], M. astracanicum [38], M. crassidens [38], M. deserti [35,52], M. duabense [40], M. incanum [50,51], M. parviflorum [42,47], M. peregrinum [44], M. persicum [53], M. thessalum [54], M. velutinum [44], M. vulgare [47,50] 24 25 β-Cubebene ST Test β-Elemene ST Train 26 Z-Caryophyllene ST 27 α-Z-Bergamotene ST 1475.610 1390 1475.506 1392 Train 1463.161 Train 1428.215 0.1 1389 0.4 1391 1.0 1407 0.1 1406 0.2 1416 0.2 - - 28 E-Caryophyllene ST Validation 1463.161 1422 24.6 1423 23.0 M. anisodon [37], M. aschersonii [34], M. astracanicum [38,46], M. crassidens [38,39], M. deserti [35,52], M. duabense [40], M. incanum [50,51], M. parviflorum [41,42,47,48], M. peregrinum [36,43,44], M. persicum [45,53], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [7–13,16,17,34,36,47,49,50] 29 β-Copaene ST Test 1459.623 1430 0.4 1430 1.3 M. incanum [50], M. vulgare [50] Plants 2021, 10, 600 4 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 30 α-E-Bergamotene ST Train 31 NI-3 - 2019 2020 Reference RIa % RIa % 1428.215 1436 0.1 1435 0.1 M. anisodon [37], M. astracanicum [46], M. crassidens [38], M. parviflorum [42,47], M. peregrinum [44], M. velutinum [44], M. vulgare [47] - 1445 0.2 1444 0.6 M. anisodon [37], M. aschersonii [34], M. astracanicum [38,46], M. crassidens [38,39], M. duabense [40], M. incanum [50,51], M. parviflorum [42,47], M. peregrinum [36,43], M. persicum [45], M. thessalum [54], M. velutinum [44], M. vulgare [8–10,12,13,15,34,36,47,50] 32 α-Humulene ST Validation 1503.999 1454 5.2 1455 5.3 33 Sesquisabinene ST Train 1442.740 - - 1457 0.9 34 E-β-Farnesene ST Test 1431.419 1457 35 C16H34 A Train 1573.436 36 NI-4 - - 37 Z-Muurola-4(14),5diene ST Train 38 NI-5 39 1.3 - 1462 1.5 1462 0.2 - Trace - 0.2 1482.433 - - 1466 0.1 - - 1469 0.1 - - - NI-6 - - 1472 0.1 - - - 40 E-Cadina-1(6),4-diene ST Train 1481.465 - - 1475 Trace M. vulgare [15] 41 γ-Muurolene ST Test 1450.203 1479 0.1 - - M. incanum [50], M. peregrinum [43,44], M. parviflorum [42], M. velutinum [44] 17.0 M. anisodon [37], M. aschersonii [34], M. astracanicum [38], M. crassidens [38,39], M. deserti [35,52], M. incanum [50,51], M. parviflorum [41,42,47,48], M. peregrinum [36,43,44], M. persicum [45,53], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [9–13,15–17,34,36,47,50] 42 43 Germacrene D ST E-β-Ionone O Test Test 1450.188 1483 9.6 1487 - M. anisodon [37], M. aschersonii [34], M. crassidens [39], M. parviflorum [41,42,47], M. peregrinum [43,44], M. persicum [45], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [8,10,12,16,17,34,36,47] - 1471.735 1486 0.4 1489 Trace M. anisodon [37], M. aschersonii [34], M. duabense [40], M. incanum [51], M. parviflorum [42], M. peregrinum [43,44], M. thessalum [54], M. vulgare [12] - 44 NI-7 - - - - 1489 0.1 45 epi-Cubebol OST Train 1622.285 - - 1495 0.2 46 Viridiflorene ST Validation 1507.447 1497 0.1 - - 47 Bicyclogermacrene ST Validation 1493.697 1498 0.2 1498 0.2 M. astracanicum [38], M. crassidens [38,39], M. deserti [35,52], M. duabense [40], M. incanum [50,51], M. parviflorum [41,42,47,48], M. peregrinum [36,43,44], M. persicum [45], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [10,11,17,50] 48 NI-8 - - - - 1499 0.7 - 49 Pentadecane A Test 1486.884 1500 0.2 1500 Trace 50 α-Muurolene ST Train 1465.650 1501 0.1 1501 0.2 M. aschersonii [34], M. deserti [35,52], M. incanum [51], M. peregrinum [43], M. velutinum [44], M. vulgare [12,13,34] 51 Germacrene A ST Train 1450.188 1508 0.1 1506 0.1 M. incanum [50], M. parviflorum [47,48] Plants 2021, 10, 600 5 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 2019 RIa 2020 % RIa % Reference 52 β-Bisabolene ST Validation 1425.139 1511 0.2 1507 0.2 M. anisodon [37], M. aschersonii [34], M. crassidens [38], M. parviflorum [47], M. peregrinum [44], M. persicum [45], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [11–13,17,34,47,49] 53 γ-Cadinene ST Test 1450.203 1513 0.2 1515 0.4 M. deserti [52], M. incanum [50], M. parviflorum [47,48], M. peregrinum [43,44], M. persicum [53], M. velutinum [44], M. vulgare [7,10,15,47] 54 δ-Cadinene ST Test 1475.070 1523 4.7 1528 9.7 M. deserti [52], M. incanum [50], M. parviflorum [42,47], M. peregrinum [43,44], M. persicum [53], M. velutinum [44], M. vulgare [7,10,15,47] 55 E-Cadina-1,4-diene ST Train 1471.521 1533 0.1 1533 0.1 M. vulgare [15] 56 α-Cadinene ST Train 1465.650 - - 1537 0.1 M. peregrinum [43,44], M. velutinum [44], M. vulgare [47] 57 α-Calacorene ST Train 1540.123 - - 1543 0.1 M. deserti [52], M. vulgare [12,15] 58 NI-9 - - 1555 0.2 1552 0.2 - 59 E-Nerolidol OST Validation 1567.136 1561 3.5 1564 1.5 M. anisodon [37], M. deserti [52], M. parviflorum [42], M. peregrinum [43,44], M. thessalum [54], M. velutinum [44], M. vulgare [9,36] 60 NI-10 - - - - 1571 0.1 - 61 NI-11 - - 1577 0.2 1575 0.9 - 62 NI-12 - - - - 1582 0.3 M. anisodon [37], M. astracanicum [46], M. crassidens [38,39], M. deserti [52], M. duabense [40], M. incanum [50,51], M. parviflorum [41,42,47,48], M. peregrinum [36,43], M. persicum [45,53], M. propinquum [41], M. thessalum [54], M. velutinum [44], M. vulgare [8–10,12,36,47,50] 63 Caryophyllene oxide OST Test 1636.612 1580 1.0 1583 1.8 64 NI-13 - - - - 1587 0.1 - 65 Viridiflorol OST Validation 1573.436 1597 0.1 - - M. aschersonii [34], M. astracanicum [38], M. crassidens [38], M. incanum [51], M. parviflorum [47], M. peregrinum [43], M. vulgare [10,12,34,47] 66 Hexadecane A Train 1594.576 1602 0.1 - - M. duabense [40], M. velutinum [44] 67 Humulene epoxide II OST Train 1626.959 1607 0.2 1607 0.2 M. anisodon [37], M. incanum [51], M. thessalum [54], M. vulgare [10] 68 Muurola-4,10(14)dien-1-β-ol OST Train 1605.330 - - 1627 0.3 69 NI-14 - - 1628 0.1 - - 70 4,4-dimethylTetracyclo [6.3.2.0(2,5).0(1,8)] tridecan-9-ol O Validation 1605.030 - - 1631 0.2 71 NI-15 - - 1632 0.1 - - 72 Caryophylla-4(12), 8(13)-dien-5-α-ol OST Train 1605.030 1636 0.1 1635 0.3 73 epi-α-Muurolol (=tau-muurolol) OST Test 1605.030 1642 0.2 1641 0.6 74 α-Muurolol (=Torreyol) OST Train 1652.148 - - 1645 0.1 - - M. astracanicum [38], M. deserti [35], M. incanum [51], M. parviflorum [42], M. peregrinum [44], M. velutinum [44] Plants 2021, 10, 600 6 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 75 α-Cadinol OST Train 76 NI-16 77 2019 2020 Reference RIa % RIa % 1682.934 1654 0.3 1654 0.9 M. crassidens [38], M. deserti [35,52], M. incanum [50,51], M. parviflorum [42], M. persicum [45], M. vulgare [12,50] - - 1658 0.2 1656 0.2 - NI-17 - - 1662 0.1 1662 0.1 - 78 E-Calamenen- 10-ol OST Train 1608.844 - - 1669 0.1 79 NI-18 - - 1668 0.2 - - - 80 NI-19 - - - - 1670 0.2 81 8-Heptadecene O Train 1607.164 - - 1673 0.2 82 1-Tetradecanol O Train 1702.771 1675 0.1 - - 83 Germacra-4(15),5, 10(14)-trien-1-α-ol OST Train 1700.003 1682 0.1 1685 0.2 84 Heptadecane A Validation 1726.886 1696 0.3 1696 0.2 M. anisodon [37], M. parviflorum [42,47], M. vulgare [10,47] 85 Pentadecanal O Validation 1581.928 1710 0.1 1711 0.1 M. anisodon [37] 86 Mint sulfide ST Train 1778.777 1733 0.1 1736 0.1 87 NI-20 - - 1734 0.1 - - - 88 NI-21 - - 1742 0.1 - - - 89 NI-22 - - 1743 0.4 1744 0.1 90 E-3-Octadecene O Train 1722.391 - - 1777 0.1 91 n-Pentadecanol O Train 1787.022 1778 0.1 - - M. parviflorum [42] 92 NI-23 - - - - 1782 0.1 - 93 Octadecane A Validation 1950.093 1796 0.1 - - M. parviflorum [47], M. peregrinum [43], M. vulgare [47] 94 NI-24 - - 1819 0.1 - - - 95 6,10,14-trimethyl2-Pentadecanone O Train 1915.818 1844 4.8 1842 0.5 M. peregrinum [44], M. velutinum [44], M. vulgare [10] 96 NI-25 - - 1849 0.1 - - - 97 NI-26 - - 1853 0.2 - - - 98 NI-27 - - 1888 0.1 - - - 99 NI-28 - - 1891 0.8 1891 0.1 - 100 Nonadecane A Test 1869.346 1897 0.2 1897 0.2 M. duabense [40], M. parviflorum [47], M. peregrinum [43], M. vulgare [10,15,47] 101 NI-29 - - 1904 0.1 1906 Trace - 102 5E,9E-Farnesyl acetone OST Train 1956.289 1916 0.3 1915 Trace M. thessalum [54], M. vulgare [15] 103 NI-30 - - 1918 Trace 1917 Trace - 104 NI-31 - - 1924 0.1 - - - 105 NI-32 - - - - 1926 Trace - 106 NI-33 - - 1925 0.1 - - - 107 NI-34 - - 1929 0.1 - - - 108 NI 1938 0.1 1940 Trace 109 Hexadecanoic acid O Validation 1995.491 1960 3.9 - - M. parviflorum [42], M. peregrinum [36], M. vulgare [36,47] 110 NI-35 - - 1973 0.1 1974 Trace - 111 Eicosane A Train 2034.560 1997 0.2 1994 0.1 M. parviflorum [48] 112 NI-36 - - 2001 0.1 - - - Plants 2021, 10, 600 7 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 113 E,E-Geranyl linalool OD Train 2028.645 114 3,7,11,15-tetramethyl(E,E)-1,6,10,14Hexadecatetraen3-ol OD 115 Manool OD Train 116 NI-37 117 NI-38 118 119 2019 2020 Reference RIa % RIa % 2027 1.6 - - - - 2028 0.9 2064.196 2057 0.3 - - - - 2061 0.1 - - - - - 2067 0.1 - - - NI-39 - - 2084 0.1 - - - NI-40 - - 2096 0.1 - - - M. aschersonii [34], M. parviflorum [42], M. vulgare [12,34] 120 Heneicosane A Train 2120.284 2101 1.6 2100 1.3 M. parviflorum [42,47], M. peregrinum [43], M. propinquum [41], M. vulgare [10] 121 NI-41 - - 2108 0.3 2105 0.2 - 122 NI-42 - - 2112 0.2 2110 0.3 - 0.4 M. anisodon [37], M. incanum [51], M. parviflorum [41,42], M. peregrinum [36], M. propinquum [41], M. vulgare [10,15] 123 Phytol OD Test 2124.818 2116 1.4 2113 124 NI-43 - - 2131 0.2 - - - 125 NI-44 - - 2143 0.2 2143 0.1 - 126 NI-45 - - 2147 0.2 - - - 127 NI-46 - - 2164 0.2 2160 0.2 - 128 NI-47 - - 2167 0.1 2172 0.3 - 129 NI-48 - - 2175 0.6 2176 0.4 - 130 NI-49 - - 2181 0.9 2179 0.2 - 131 NI-50 - - 2183 0.4 - - - 132 NI-51 - - 2198 2.4 2195 2.4 - 133 Docosane A Validation 2194.421 2205 0.9 2198 0.6 M. crassidens [39], M. parviflorum [47] 134 NI-52 - - - - 2201 0.1 - 135 NI-53 - - - - 2209 0.3 - 136 NI-54 - - 2215 0.3 - - - 137 NI-55 - - 2225 0.3 2221 0.1 - 138 NI-56 - - 2246 0.3 - - - 139 NI-57 - - 2258 0.2 2253 0.3 - 140 NI-58 - - 2270 0.1 2265 0.1 - 141 NI-59 - - 2277 0.2 2274 0.2 - 142 NI-60 - - 2293 3.8 2288 1.7 143 Tricontane A Train 2381.642 2305 3.6 2302 2.6 144 NI-61 - - 2309 0.2 2305 0.2 - 145 NI-62 - - 2344 0.2 2341 0.3 - 146 NI-63 - - 2380 0.1 2377 0.1 - 147 NI-64 - - 2383 0.1 2382 0.1 - 148 Tetracosane A Train 2493.491 2401 0.3 2395 0.2 M. deserti [52], M. parviflorum [41,42,47], M. propinquum [41] 149 NI-65 - - 2454 0.4 2447 0.2 - 150 NI-66 - - 2488 0.2 2483 0.2 - 151 Pentacosane A Test 2510.087 2503 0.8 2497 0.6 M. anisodon [37], M. parviflorum [42,47] Plants 2021, 10, 600 8 of 17 Table 1. Cont. No Compound/Class Cycle RIpred. 152 Heptacosane A Train 153 NI-67 - 154 Octacosane A 155 Squalene T Train 2019 2020 Reference RIa % RIa % 2730.537 2702 0.6 2696 0.5 - - - 2766 0.1 M. crassidens [39], M. parviflorum [42], M. persicum [45] 2801 0.1 2791 Trace 2870.673 2835 0.1 2823 0.1 M. anisodon [37], M. aschersonii [34], M. incanum [51], M. parviflorum [42], M. vulgare [34] 156 NI-68 - - 2868 0.1 2855 0.1 - 157 Nonacosane A Test 2930.732 2905 0.7 2892 0.6 M. anisodon [37], M. crassidens [39], M. persicum [45] 158 Untriacontane A Validation 3150.673 3105 0.4 3095 0.3 159 NI-69 - - - - 3212 0.1 160 Tritriacontane A Train 3319.753 3300 0.1 3301 Trace Oxygenated monoterpenes OMN Sesquiterpene hydrocarbons ST Oxygenated sesquiterpenes OST Oxygenated diterpenes OD Triterpene T Aromatics AR Alkanes A Other O NI Total 0.1 0.2 52.0 67.8 5.8 6.2 3.3 1.3 0.1 0.4 11.7 9.9 16.7 100 0.1 0.4 7.4 3.4 12.5 99.3 - RIpred. —BRT calculated retention index; RIa —retention index experimentally obtained on a HP-5MS column; Other—aliphatic hydrocarbons, aliphatic aldehydes and alcohols, aliphatic acids, their esters and aldehydes, aromatic esters with aliphatic acids, alkyl-aromatic alcohols, or aryl esters of aromatic acids; NI—not identified compound. The predicted RIs are shown in Table 1, and confirm the good quality of the constructed BRT model by showing the relationship between the predicted and experimental RI values. Graphical comparison between experimentally obtained RIs of M. vulgare volatiles composition (RIa ), the retention time indices found in NIST database (RIb ) and the retention time indices predicted by the two BRT models (RIpred. ) are presented in Figure 1. Figure 1. Retention indices (RIs) of the M. vulgare volatiles composition, from experimentally obtained GC–MS data on a HP-5MS column (RIa ) and NIST database (RIb ). Plants 2021, 10, 600 9 of 17 In order to calculate the molecular descriptors, the PaDel-descriptor was used in this investigation. Due to a great amount of data that was obtained, it was required to select the most important set of descriptors to build the adequate model which would be able to predict the RIs [55]. The factor analysis was done before the GA calculation, and only ca. 320 uncorrelated descriptors remained in the GA calculation [56,57]. The seven most significant molecular descriptors chosen by GA are as follows: four 2D autocorrelation descriptors (AATSC4e, AATSC2p, GATS6v and MATS5v), two Barysz matrix descriptors (VR1_Dzs and SM1_Dzv) and Vertex adjacency information (magnitude) descriptor (VAdjMat). The predicted RIs and molecular descriptors are presented in Table 1. Seven molecular descriptors were utilized for predictions of RIs in the two BRT models. The predicted RIs are presented in Figure 2, and visually confirm the adequate prediction capabilities of the constructed BRT by showing the relationship between the predicted and experimental retention values. Figure 2. Comparison of experimentally obtained RIs on a HP-5MS column (RIa) with BRT pre-dicted values (RIpred.) in 2019 (a) and 2020 (b). Separation of compounds in GC–MS and their RIs is linked to their affinity towards mobile and stationary phases. Affinity and solubility of separated molecules directly depend on their chemical structure and physico-chemical properties, which could be expressed by molecular descriptors. According to Pearson’s correlation coefficients, there was a rather poor correlation between all 3D autocorrelation descriptors (Table 2). Therefore, utilized molecular descriptors were appropriate to predict RIs of compounds in M. vulgare by the two multivariate BRT models [58]. Table 2. The correlation coefficient matrix for the selected descriptors by GA. AATSC4e AATSC2p MATS5v GATS6v SM1 Dzv VR1 Dzs AATSC2p MATS5v GATS6v SM1_Dzv VR1_Dzs VAdjMat 0.031 −0.138 −0.265 −0.135 −0.131 0.212 0.030 0.036 −0.008 0.072 0.205 −0.231 0.066 0.131 0.058 0.224 −0.010 0.109 0.214 0.084 2.339 Detailed explanations about the descriptors were found in the Handbook of Molecular Descriptors [59]. These descriptors encode different aspects of the molecular structure and were applied to develop the QSRR model. According to Pearson’s correlation, there was a rather poor correlation between all molecular descriptors. Hence, utilized descriptors were appropriate to predict RIs of compounds isolated from M. vulgare volatiles by the two multivariate BRT models. The calibration and predictive capability of a QSRR model Plants 2021, 10, 600 10 of 17 should be tested through model validation. The most widely used squared correlation coefficient (r2 ) can provide a reliable indication of the fit of the model, thus, it was employed to validate the calibration capability of a QSRR model. In order to explore the nonlinear relationship between RIs and the descriptors selected by GA, BRT technique was used to build the two predictive models. Two BRT models were constructed to predict the retention time of compounds isolated from M. vulgare volatiles, respectively. The coefficients of determination were 0.956 and 0.964, respectively, indicating that these models could be used for prediction of RIs, due to low prediction error and high r2 . The tests of the two BRT models fit (2019 and 2020) are shown in Table 3, with the higher r2 values and lower χ2 , MBE, RMSE, and MPE values showing the better fit to the experimental results [60,61]. Table 3. The “goodness of fit” tests for the developed BRT model. Boosted Tree Model χ2 RMSE MBE MPE r2 2019 2020 4455.272 3975.751 66.160 62.581 −13.063 −7.698 3.285 3.241 0.956 0.964 χ2 —reduced chi-square, MBE—mean bias error, RMSE—root mean square error, MPE—mean percentage error. Obtained results reveal the reliability of the BRT models for predicting the RIs of compounds in M. vulgare volatiles obtained by GC–MS analysis. The influence of the seven most important molecular descriptors, identified by using genetic algorithm on the RIs was studied in this section. According to the Figure 3, VAdjMat was the most important molecular descriptor for chemical compounds’ RIs calculation in M. vulgare, during 2019, while VR1 Dzs was the most important variable during 2020. Figure 3. Predictor importance of the molecular descriptors on RI in 2019 (a) and 2020 (b). 3. Discussion According to the cluster analysis (unrooted cluster tree) with 37 samples of Marrubium sp. volatiles from literature and average values from this study (Figure 4), it could be said that there are several chemotypes, but only E-caryophyllene chemotype [9,12,36,38,47,50,51] is clearly segregated. However, these are samples of M. vulgare, M. incanum, M. parviflorum, M. peregrinum, and M. crassidens grown in Serbia, Poland, Slovakia, Egypt, and Iran. This indicated that genus Marrubiumis very diverse in the case of volatiles composition. α β β Plants 2021, 10, 600 11 of 17 Figure 4. Unrooted cluster tree for different Marrubium samples. In addition, E-caryophyllene is a compound which is occurring in all samples (except M. vulgare from Eastern Algeria [15]), but in E-caryophyllene chemotype its content ranges between 15.6% and 45.8%. Other chemotypes can be classified as β-bisabolene (13.1–28.3%) [11,17,34,47], α-pinene (21.5–28.9%) [16,53], β-farnesene (20.2–24.2%) [37,44], E-caryophyllene + caryophyllene oxide chemotype [44,46,54], and diverse (unclassified) chemotypes [7,8,10,13,15,35,38,40,41,43,45,49,52]. Occurring polymorphism could be a consequence of adaptation to grow in different environments [19,62], especially ecological conditions such as humidity, temperature and altitude [22] as well as hybridization [20] strongly affected the chemotypes, as well as biotic and abiotic stresses (including temperature, light, water, salt, and oxidative stresses) [63]. Detected compounds in M. vulgare volatiles obtained by GC–MS analysis were used for QSRR analysis. The following seven molecular descriptors that characterize the RIs of obtained compounds were suggested by the genetic algorithm. Selected molecular descriptors were not autocorrelated which was suggested by a correlation coefficient matrix; thus, descriptors were suitable for QSRR analysis. These descriptors were utilized as inputs for the boosted trees regression models, for estimating the RIs using a set of GC– MS data from a series of 160 compounds found in M. vulgare volatiles. Statistical models that quantify the relation between the structure of molecules and their chromatographic RIs were represented by the quantitative structure retention relationship (QSRR) model [58,64]. č Numerous publications related to the QSRR analysis in plants from Lamiaceae family could be found in the literature: Thymus vulgaris [65], T. serpyllum [66], Satureja kitaibelii [55], Salvia officinalis [67], as well as Stachys sp. [68]. The connection between the molecular Plants 2021, 10, 600 12 of 17 descriptors and the retention time can be established by artificial neural network, machine learning algorithms [69–73] or by boosted trees regression (BRT) [74]. 4. Materials and Methods 4.1. Plant Material M. vulgare was grown in the Institute of Field and Vegetable Crops (IFVCNS) collection garden of medicinal and aromatic plants in Bački Petrovac (45◦ 21′ N; 19◦ 35′ E), confirmed by Milica Rat, PhD, and deposited at the Herbarium BUNS (Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad) under the voucher number 2-1409. After seed maturation (August 2018), it was collected and sown in field conditions in September 2018 and 2019. The experimental plot was 10 m long and 5 m wide, with a 70 cm spacing between rows. From the seven rows, only three central rows were used for collecting plant material to avoid edge effects (one row one sample). 4.2. Volatiles Isolation and Analysis Flowering aerial parts of M. vulgare (Marrubii herba) were collected during July 2019 and 2020, dried in a solar dryer at a temperature of 40◦ with air circulation. After drying, plant material was fragmented, and volatiles was extracted by Clevenger apparatus. Taking in account that M. vulgare produces trace amounts of essential oil, it was trapped in nhexane. This process was performed in tree repetition for both years, as well as analysis of chemical composition. GC–MS analysis was carried out using an Agilent 7890A apparatus equipped with a 5975 C MSD, FID and a nonpolar HP-5MS fused-silica capillary column (30 m × 0.25 mm, film thickness 0.25 µm). The carrier gas was helium, and its inlet pressure was 19.6 psi and linear velocity of 1 mL/min at 210 ◦ C. The injector temperature was 250 ◦ C, injection volume was 1 µL, split ratio, 10:1. Mass detection was carried out under source temperature conditions of 230 ◦ C and interface temperature of 315 ◦ C. The EI mode was set at electron energy, 70 eV with mass scan range of 40–600 amu. Temperature was programmed from 60 to 300 ◦ C at a rate of 3 ◦ C/min. The components were identified based on their linear retention index relative to C8-C32 n-alkanes, compared with data reported in the literature (Adams4 and NIST11 databases). The relative percentage of the oil constituents was expressed as percentages by FID peak area normalization. 4.3. QSRR Analysis PaDel-descriptor software was used to calculate specified molecular descriptors [75], as described in our previous investigation [66]. Factor analysis and genetic algorithm (GA) were used to determine the most important descriptors [76,77]. The relationship between the chosen descriptors was examined and collinear descriptors were excluded. Statistica 10 software was used for the statistical investigation of the data [78]. 4.4. BRT Model In order to relate and to predict categorical or continuous dependent variables the BRT model could be used [79,80], as it does not require transformation or outliers [81]. The BRT method calculation is connected to the boosting methods enforced to regression trees [82]. The main idea is to calculate a set of simple trees, where each successive tree is built for the prediction residuals of the preceding tree [83]. This method builds binary trees such as partition the data into two samples at each split node [78]. The decision trees are combined through a cross-validation or “boosting” procedure in order to acquire the single computational model [84]. BRT modeling consists of the following steps: (a) an initial regression tree is defined according to a minimum loss function; (b) the other trees are engaged in the iterative process in which several new regression trees were developed and selected to the subsequent according to the StatSoft Statistica’s recommendation—the least square error (LSE); (c) step (b) is repeated until a stopping criterion is reached (for instance, the value of LSE). Plants 2021, 10, 600 13 of 17 In this study, several regularization parameters were set in order to optimize the fit between experimental results and computing model: the number of trees (between 100 and 1000), learning rate (between 0.0005 and 0.1), random test data proportion (0.1–0.9) and subsample proportion (0.1–0.9). According to Statistica’s manual, prior to computation, a subsample of data is created, according to random test data proportion of the cases, and these data are treated as test samples used to evaluate the appropriate fit of the model. The remaining set of data is used for the analyses via stochastic gradient boosting (for the selection of samples for consecutive boosting steps). 4.5. Cluster Analysis The cluster analysis (CA) was used to evaluate intra- and interpopulation variability and differentiation of volatile constituents of Marrubium samples collected in different locations and/or taken from literature reports. The phylogenetic tree diagram for Marrubium samples was calculated and plotted using R software 4.0.3 (64-bit version). The R package “ape” (Analysis of Phylogentics and Evolution) was used for calculation, applied as a graphical tool to represent the arrangements of similar volatiles concentration (evaluated in the cluster analysis). The obtained experimental results were collected in the matrix, after which the hierarchical cluster analysis was performed. The distance matrix was determined using the Euclidean method, while the cluster analysis was performed using the “complete” method. 5. Conclusions The main components in M. vulgare volatiles were E-caryophyllene (24.6% and 23.0%), followed by germacrene D (9.6% and 17.0%), α-humulene (5.2% and 5.3%) and α-copaene (3.3% and 6.1%) in 2019 and 2020, respectively. All these compounds are from sesquiterpene hydrocarbons class, which was dominant in both years of the investigation, 52.0% in 2019 and 67.8% in 2020. The results demonstrated that the boosted trees regression models were adequate in predicting the RIs of the compounds in M. vulgare volatiles obtained by GC–MS analysis on a HP-5MS column. The coefficients of determination were 0.956 and 0.964 (for compounds found in M. vulgare volatiles, during the years 2019 and 2020, respectively), which is a good indication that these models could be used as a fast mathematical tool for prediction of RIs, due to low prediction error and moderately high r2 . Suitable models with high statistical quality and low prediction errors were derived, and it could be further used for estimation of RIs of newly detected compounds. According to the unrooted cluster tree with 37 samples of Marrubium sp. volatiles from literature and average values from this study, it could be said that there are several chemotypes: E-caryophyllene, β-bisabolene, α-pinene, β-farnesene, E-caryophyllene + caryophyllene oxide chemotype, and diverse (unclassified) chemotypes. However, occurring polymorphism could be a consequence of adaptation to grow in different environments, especially ecological conditions such as humidity, temperature and altitude, as well as hybridization which strongly affected the chemotypes. Further research on M. vulgare chemotypes needs to be focused on genetic markers, because evaluation of genetic diversity has key importance in improving the quality of raw material used for industrial purposes. Author Contributions: Conceptualization, M.A. and J.O.; methodology, S.I. and K.S.; software, L.P.; validation, S.I., K.S. and T.Z.; formal analysis, L.P.; investigation, M.A.; resources, M.A. and V.S.; data curation, L.P.; writing—original draft preparation, M.A.; writing—review and editing, T.Z. and V.S.; visualization, L.P.; supervision, J.O.; project administration, T.Z.; funding acquisition, M.A., T.Z., J.O. and V.S. All authors have read and agreed to the published version of the manuscript. Funding: This research was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia, grant number: 451-03-9/2021-14/200032. Data Availability Statement: Not applicable. Plants 2021, 10, 600 14 of 17 Acknowledgments: We thank Vele Tešević, Marina Todosijević, Jovana Stanković Jeremić, Jovana Ljujić, and Mirjana Cvetković for participating in this research. Conflicts of Interest: The authors declare no conflict of interest. References 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. Yabrir, B. Essential oil of Marrubium vulgare: Chemical composition and biological activities. A review. Nat. Prod. Sci. 2019, 25, 81–91. [CrossRef] Lippai, A.; Smith, P.A.; Price, T.V.; Weiss, J.; Lloyd, C.J. Effects of temperature and water potential on germination of horehound (Marrubium vulgare) seeds from two Australian Localities. Weed Sci. 1996, 44, 91–99. [CrossRef] Nedjimi, B.; Souissi, Z.E.; Guit, B.; Daoud, Y. Differential effects of soluble salts on seed germination of Marrubium vulgare L. J. Appl. Res. Med. Aromat. Plants 2020, 17, 100250. [CrossRef] Dmitruk, M.; Haratym, W. Morphological differentiation of non-glandular and glandular trichomes on Marrubium vulgare L. Mod. Phytomorphol. 2014, 6, 85. Aćimović, M.; Jeremić, K.; Salaj, N.; Gavarić, N.; Kiprovski, B.; Sikora, V.; Zeremski, T. Marrubium vulgare L.: A phytochemical and pharmacological overview. Molecules 2020, 25, 2898. [CrossRef] Lodhi, S.; Vadnere, G.P.; Sharma, V.K.; Usman, M.R. Marrubium vulgare L.: A review on phytochemical and pharmacological aspects. J. Intercult. Ethnopharmacol. 2017, 6, 429–452. [CrossRef] Mahmoud, A.A.; Gendy, A.S.H.; Said-Al Ahl, H.A.H.; Grulova, D.; Astatkie, T.; Abdelrazik, T.M. Impact of harvest time and water stress on the growth and essential oil components of horehound (Marrubium vulgare). Sci. Hortic. 2018, 232, 139–144. [CrossRef] Said-Al Ahl, H.A.H.; Gendy, A.S.H.; Mahmoud, A.A.; Mohamed, H.F.Y. Essential oil composition of Marrubium vulgare L. cultivated in Egypt. Int. J. Plant. Sci. Ecol. 2015, 1, 138–141. El-Leithy, A.S.; El-Hanafy, S.H.; Omer, E.A.; El-Sayed, A.A.A. Effect of nitrogen and potassium biofertilization on growth, yield and essential oil production of the white horehound, Marrubium vulgare L. plant. J. Hortic. Sci. Ornam. Plants 2013, 5, 46–59. Weel, K.G.C.; Venskutonis, P.R.; Pukalskas, A.; Gruzdiene, D.; Linssen, J.P.H. Antioxidant activity of horehound (Marrubium vulgare L.) grown in Lithuania. Lipid 1999, 101, 395–400. [CrossRef] Morteza-Semnani, K.; Saeedi, M.; Babanezhad, E. The essential oil composition of Marrubium vulgare L. from Iran. J. Essent Oil Res. 2008, 20, 488–489. [CrossRef] Golparvar, A.R.; Hadipanah, A.; Mehrabi, A.M.; Armin, A. Essential oil composition of Marrubium vulgare L. from Iran. Flavour Herb. Drug 2015, 6, 1–5. Zarai, Z.; Kadri, A.; Chobba, I.B.; Mansour, R.B.; Bekir, A.; Mejdoub, H.; Gharsallah, N. The in-vitro evaluation of antibacterial, antifungal and cytotoxic properties of Marrubium vulgare L. essential oil grown in Tunisia. Lipids Hum. Health Dis. 2011, 10, 161. [CrossRef] Kadri, A.; Zarai, Z.; Bekir, A.; Gharsallah, N.; Damak, M.; Gdoura, R. Chemical composition and antioxidant activity of Marrubium vulgare L. essential oil from Tunisia. Afr. J. Biotechnol. 2011, 10, 3908–3914. Abadi, A.; Hassani, A. Essential oil composition and antioxidant activity of Marrubium vulgare L. growing wild in Eastern Algeria. Int. Lett. Chem. Phys. Astron. 2013, 9, 17–24. [CrossRef] Bayir, B.; Gunduz, H.; Usta, T.; Sahin, E.; Ozdemir, Z.; Kayir, O.; Sen, O.; Aksit, H.; Elmastas, M.; Erenler, R. Chemical composition of essential oil from Marrubium vulgare L. leaves. J. New Results Sci. 2014, 6, 44–50. El-Hallous, E.I.; Alsanie, W.F.; Ismail, I.A.; Dessoky, E.S. Utilization of Marrubium vulgare extract as a therapeutic to hepatic damage induced by Carbon Tetrachloride in rats. Int J. Pharm. Res. Allied Sci. 2018, 7, 168–178. Yabrir, B. Chemical composition and biological activities of some Marrubium species essential oil: A review. Chem. J. Mold. 2018, 13, 8–23. [CrossRef] Salehi, N.; Kharazian, N.; Shiran, B. Genetic diversity of Marrubium species from Zagros Region (Iran), using inter simple sequence repeat molecular marker. J. Sci. Islamic Repub. Iran. 2018, 29, 7–19. Kharazian, N.; Hashemi, M. Chemotaxonomy and morphological studies in five Marrubium L. species in Iran. Iran. J. Sci. Technol. Trans. Sci. 2017, 41, 17–31. [CrossRef] Akgul, G.; Ketenoglu, O.; Pinar, N.M.; Kurt, L. Pollen and seed morphology of the genus Marrubium (Lamiaceae) in Turkey. Ann. Bot. Fenn. 2008, 45, 1–10. [CrossRef] Ahvazi, M.; Jamzad, Z.; Balali, G.R.; Saeidi, H. Trichome micro-morphology in Marrubium L. (Lamiaceae) in Iran and the role of environmental factors on their variation. Iran. J. Bot. 2016, 22, 39–58. [CrossRef] Ahvazi, M.; Balali, G.R.; Jamzad, Z.; Saeidi, H. A taxonomical, morphological and pharmacological review of Marrubium vulgare L., an old medicinal plant in Iran. J. Med. Plants 2018, 17, 7–24. Martin, E.; Cetin, O.; Akgul, G.; Hilal, A.Y. New chromosome numbers in the genus Marrubium (horehound) from Turkey. Biol. Divers. Conserv. 2011, 4, 185–188. Buyukkartal, H.N.; Colgecen, H.; Akgul, G. Comparative leaf, stem and root anatomies of taxa Marrubium bourgaei and Marrubium heterodon (Lamiaceae). Aust. J. Crop Sci. 2016, 10, 1516–1522. [CrossRef] Marzouk, R.I.; El-Darier, S.M.; Nour, I.H.; Kamal, S.A. Numerical taxonomic study of Marrubium L. (Lamiaceae) in Egypt. Catrina 2015, 13, 25–35. Plants 2021, 10, 600 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. 37. 38. 39. 40. 41. 42. 43. 44. 45. 46. 47. 48. 49. 50. 51. 52. 53. 54. 55. 15 of 17 Sgarbossa, J.; Schmidt, D.; Schwerz, F.; Schwerz, L.; Prochnow, D.; Caron, B.O. Effect of season and irrigation on the chemical composition of Aloysia triphylla essential oil. Rev. Ceres 2019, 66, 85–93. [CrossRef] Castelo, A.V.M.; Del Menezzi, C.H.S.; Resck, I.S. Seasonal variation in the yield and the chemical composition of essential oils from two Brazilian native arbustive species. J. Appl. Sci. 2012, 12, 753–760. [CrossRef] Yavari, A.; Nazeri, V.; Sefidkon, F.; Hassani, M.E. Influence of some environmental factors on the essential oil variability of Thymus migricus. Nat. Prod. Comm 2010, 5, 943–948. [CrossRef] Zouari-Bouassida, K.; Trigui, M.; Makni, S.; Jlaiel, L.; Tounsi, S. Seasonal variation in essential oils composition and the biological and pharmaceutical protective effects of Mentha longifolia leaves grown in Tunisia. Biomed. Res. Int. 2018, 7856517. [CrossRef] Gosztola, B.; Sarosi, S.; Nemeth, E. Variability of the essential oil content and composition of chamomile (Matricaria recutita) affected by weather conditions. Nat. Prod. Commun. 2010, 5, 465–470. [CrossRef] Acimovic, M.; Cvetkovic, M.; Stankovic, J. Effect of weather conditions, location and fertilization on coriander fruit essential oil quality. Essent Oil Bear Plants 2016, 19, 1208–1215. Aziz, E.E.; Badawy, E.M.; Zheljazkov, V.D.; Nicola, S.M.; Fouad, H. Yield and chemical composition of essential oil of Achillea millefolium L. as affected by harvest time. Egypt J. Chem. 2019, 62, 533–540. [CrossRef] Hamdaoui, B.; Wannes, W.A.; Marrakchi, M.; Brahim, N.B.; Marzouk, B. Essential oil composition of two Tunisian horehound species: Marrubium vulgare L. and Marrubium aschersonii Magnus. J. Essent Oil Bear Plants 2013, 16, 608–612. [CrossRef] Laouer, H.; Yabrir, B.; Djeridane, A.; Yousfi, M.; Beldovini, N.; Lamamra, M. Composition, antioxidant and antimicrobial activities of the essential oil of Marrubium deserti. Nat. Prod. Commun. 2009, 4, 1133–1138. [CrossRef] [PubMed] Nagy, M.; Svajdlenka, E. Comparison of essential oils from Marrubium vulgare L. and M. peregrinum L. J. Essent Oil Res. 1998, 10, 585–587. [CrossRef] Kırımer, N.; Kurkcuoglu, M.; Akgul, G.; Baser, K.H.H.; Mahmoud, A.A. Composition of the essential oil of Marrubium anisodon C. Koch of Turkish origin. Rec Nat. Prod. 2015, 9, 234–236. Teimori, M.; Khavari-Nejad, R.A.; Yassa, N.; Nejadsatari, T. Analysis of the essential oil of Marrubium crassidens Bioos. and M. astracanicum Jacq. J. Appl. Sci. 2008, 8, 1793–1795. [CrossRef] Hamedeyazdan, S.; Asnaashari, S.; Fathiazad, F. Characterization of non-terpenoids in Marrubium crassidens Bioss. essential oil. Adv. Pharm. Bull. 2013, 3, 429–432. Golmakani, H.; Rabbani Nasab, H.; Sharifan, M.; Kamali, H.; Yadollahi, A. The essential oil composition and antibacterial activity of Marrubium dubanese Murata from North Khorassan Province, Iran. J. Essent Oil Bear Plants 2016, 19, 963–971. [CrossRef] Hamedeyazdan, S.; Zarei, M.; Salem, A.; Asnaashari, S.; Fathiazad, F. Essential oil of two Iranian horehound species: Marrubium propinquum and Marrubium parviflorum. Pharm. Sci. 2017, 23, 143–149. [CrossRef] Bal, Y.; Kaban, S.; Kirimer, N.; Baser, K.H.C. Composition of the essential oil of Marrubium parviflorum Fisch. et Mey. subsp. oligodon (Boiss.) Seybold. J. Essent Oil Res. 1999, 11, 300–302. [CrossRef] Kaurinovic, B.; Vlaisavljevic, S.; Popovic, M.; Vastag, D.; Djurendic-Brensel, M. Antioxidant properties of Marrubium peregrinum L. (Lamiaceae) essential oil. Molecules 2010, 15, 5943–5955. [CrossRef] Lazari, D.; Skaltsa, H.; Constantinidis, T. Essential oils of Marrubium velutinum Sm. and Marrubium peregrinum L., growing wild in Greece. Flavour Fragr. J. 1999, 14, 290–292. [CrossRef] Hamedeyazdan, S.; Fathiazad, F.; Asnaashari, S. Chemical composition of the essential oil from Marrubium persicum C.A. Mey. (Lamiaceae). Pharm. Sci. 2013, 19, 35–38. Nik, B.Z.; Mirza, M. Composition of the essential oil of Marrubium astracanicum Jacq. J. Essent Oil Res. 2003, 15, 342–343. [CrossRef] Khanavi, M.; Ghasemian, L.; Motlagh, E.H.; Hadjikhoondi, A.; Shafiee, A. Chemical composition of the essential oils of Marrubium parviflorum Fisch. & C.A. Mey. and Marrubium vulgare L. from Iran. Flavour Fragr. J. 2005, 20, 324–326. Sarikurkcu, C.; Ozer, M.S.; Calli, N.; Popović-Djord̄ević, J. Essential oil composition and antioxidant activity of endemic Marrubium parviflorum subsp. oligodon. Ind. Crops Prod. 2018, 119, 209–213. [CrossRef] Miloudi, K.; Hamimed, A.; Benmimoun, Y.; Bellebna, Y.; Taibi, A.; Tilmatine, A. Intensification of essential oil extraction of the Marrubium vulgare using pulsed electric field. J. Essent Oil Bear Plants 2018, 21, 811–824. [CrossRef] Zawislak, G. Comparison of chemical composition of the essential oil from Marrubium vulgare L. and M. incanum Desr. during the second year of cultivation. Acta Agrobot. 2015, 68, 59–62. [CrossRef] Petrovic, S.; Pavlovic, M.; Maksimovic, Z.; Milenkovic, M.; Couladis, M.; Tzakou, O.; Niketic, M. Composition and antimicrobial activity of Marrubium incanum Desr. (Lamiaceae) essential oil. Nat. Prod. Commun. 2009, 4, 431–434. [CrossRef] [PubMed] Chemsa, A.E.; Zellagui, A.; Ozturk, M.; Erol, E.; Ceylan, O.; Duru, M.E.; Gherraf, N. Antibiofilm formation, antioxidant and anticholinesterase activities of essential oil and methanol extract of Marrubium deserti de Noe. J. Mater. Env. Sci. 2016, 7, 993–1000. Mohammadhosseini, M. First report of screening of the profiles of the essential oils and volatiles from the aerial parts of Marrubium persicum using classical and advanced methods prior to Gas Chromatographic Mass Spectrometric determination. J. Med. Plants Prod. 2016, 2, 169–180. Agryropoulou, C.; Skaltsa, H. Identification of essential oil components of Marrubium thessalum Boiss. & Heldr., growing wild in Greece. Nat. Prod. Res. 2012, 26, 1–7. Aćimović, M.; Pezo, L.; Tešević, V.; Čabarkapa, I.; Todosijević, M. QSRR Model for predicting retention indices of Satureja kitaibelii Wierzb. Ex Heuff. essential oil composition. Ind. Crops Prod. 2020, 154, 112752. [CrossRef] Plants 2021, 10, 600 56. 57. 58. 59. 60. 61. 62. 63. 64. 65. 66. 67. 68. 69. 70. 71. 72. 73. 74. 75. 76. 77. 78. 79. 80. 81. 82. 16 of 17 Nekoei, M.; Salimi, M.; Dolatabadi, M.; Mohammadhosseini, M. Prediction of antileukemia activity of berbamine derivatives by genetic algorithm–multiple linear regression. Mon. Chem. 2011, 142, 943. [CrossRef] Nekoei, M.; Mohammadhosseini, M.; Pourbasheer, E. QSAR study of VEGFR-2 inhibitors by using genetic algorithm-multiple linear regressions (GA-MLR) and genetic algorithm-support vector machine (GA-SVM): A comparative approach. Med. Chem. Res. 2015, 24, 3037–3046. [CrossRef] Azar, P.A.; Nekoei, M.; Riahi, S.; Ganjali, M.R.; Zare, K. A quantitative structure-retention relationship for the prediction of retention indices of the essential oils of Ammoides atlantica. J. Serb. Chem Soc. 2011, 76, 891–902. [CrossRef] Todeschini, R.; Consonni, V. Handbook of Molecular Descriptors, Methods and Principles in Medicinal Chemistry; Wiley-VCH Verlag GmbH: Weinheim, Germany, 2000. Bakić, V.V.; Pezo, M.L.; Jovanović, M.P.; Turanjanin, V.M.; Vučićević, B.S.; Mirkov, N.S. Technical analysis of photovoltaic/wind systems with hydrogen storage. Science 2012, 16, 865–875. Arsenović, M.; Pezo, L.; Stanković, S.; Radojević, Z. Factor space differentiation of brick clays according to mineral content: Prediction of final brick product quality. Appl. Clay. Sci. 2015, 115, 108–114. [CrossRef] Boulila, A.; Sanaa, A.; Salem, I.B.; Rokbeni, N.; Mrabet, Y.; Hosni, K.; Fernandez, X. Antioxidant properties and phenolic variation in wild populations of Marrubium vulgare L. (Lamiaceae). Ind. Crops Prod. 2015, 76, 616–622. [CrossRef] Mahdavi, A.; Moradi, P.; Mastinu, A. Variation in terpene profiles of Thymus vulgaris in water deficit stress response. Molecules 2020, 25, 1091. [CrossRef] [PubMed] Noorizadeh, H.; Farmany, A.; Noorizadeh, M. Quantitative structure-retention relationships analysis of retention index of essential oils. Química Nova 2011, 34, 242–249. [CrossRef] Driouche, Y.; Messadi, D. Quantitative structure-retention relationship model for predicting retention indices of constituents of essential oils of Thymus vulgaris (Lamiaceae) (Short communication). J. Serb. Chem Soc. 2019, 84, 405–416. [CrossRef] Acimovic, M.; Pezo, L.; Stankovic Jeremic, J.; Cvetkovic, M.; Rat, M.; Cabarkapa, I.; Tesevic, V. QSRR model for predicting retention indices of geraniol chemotype of Thymus serpyllum essential oil. J. Essent Oil Bear Plants 2020, 23, 464–473. [CrossRef] Pavlić, B.; Teslić, N.; Kojić, P.; Pezo, L. Prediction of the GC-MS retention time for terpenoids detected in sage (Salvia officinalis L.) essential oil using QSRR approach. J. Serb. Chem. Soc. 2020, 85, 9–23. [CrossRef] Noorizadeh, H. Linear and nonlinear quantitative structure linear retention indices relationship models for essential oils. Eurasian J. Anal. Chem 2013, 8, 50–63. Héberger, K. Quantitative structure–(chromatographic) retention relationships. J. Chromatogr A 2007, 1158, 273–305. [CrossRef] Kaliszan, R.; Baczek, ˛ T.; Buciński, A.; Buszewski, B.; Sztupecka, M. Prediction of gradient retention from the linear solvent strength (LSS) model, quantitative structure-retention relationships (QSRR), and artificial neural networks (ANN). J. Sep. Sci 2003, 26, 271–282. [CrossRef] Khodadoust, S.; Ghaedi, M.; Hadjmohammadi, M.R. Dispersive nano solid material-ultrasound assisted microextraction as a novel method for extraction and determination of bendiocarb and promecarb: Response surface methodology. Talanta 2013, 116, 637–646. [CrossRef] Wolfender, J.L.; Martia, G.; Thomas, A.; Bertranda, S. Current approaches and challenges for the metabolite profiling of complex natural extracts. J. Chromatogr. A 2015, 1382, 136–164. [CrossRef] [PubMed] Zisi, C.; Sampsonidis, I.; Fasoula, S.; Papachristos, K.; Witting, M.; Gika, H.G.; Nikitas, P.; Pappa-Louisi, A. QSRR modeling for metabolite standards analyzed by two different chromatographic columns using multiple linear regression. Metabolites 2017, 7, 7. [CrossRef] Deconinck, E.; Zhang, M.H.; Petitet, F.; Dubus, E.; Ijjaali, I.; Coomans, D.; Vander Heydena, Y. Boosted regression trees, multivariate adaptive regression splines and their two-step combinations with multiple linear regression or partial least squares to predict blood–brain barrier passage: A case study. Anal. Chim. Acta 2008, 609, 13–23. [CrossRef] Yap, C.W. PaDEL-descriptor: An open source software to calculate molecular descriptors and fingerprints. J. Comput. Chem. 2011, 32, 1446–1474. [CrossRef] Goldberg, D.E. Genetic Algorithms in Search, Optimisation and Machine Learning; Addison-Wesley: Boston, MA, USA, 1989. Gramatica, P. Principles of QSAR models validation: Internal and external. Qsar Comb. Sci 2007, 26, 694–701. [CrossRef] Statistica 10 Software (StatSoft, Inc. STATISTICA, ver. 10, Data Analysis Software System). Available online: https://www. statsoft.de/en/software/tibco-statisticatm (accessed on 15 December 2018). Foroughia, M.; Hossein, M.; Azqhandic, A.; Kakhki, S. Bio-inspired, high, and fast adsorption of tetracycline from aqueous media using Fe3O4-g-CN@PEI-β-CD nanocomposite: Modeling by response surface methodology (RSM), boosted regression tree (BRT), and general regression neural network (GRNN). J. Hazard. Mater. 2020, 388, 121769. [CrossRef] Cheong, Y.L.; Leitão, P.J.; Lakes, T. Assessment of land use factors associated with dengue cases in Malaysia using Boosted Regression Trees. Spat. Spatiotemporal Epidemiol. 2014, 10, 75–84. [CrossRef] [PubMed] Froeschke, J.T.; Froeschke, B.F. Spatio-temporal predictive model based on environmental factors for juvenile spotted seatrout in Texas estuaries using boosted regression trees. Fish. Res. 2011, 111, 131–138. [CrossRef] Sut, N.; Simsek, O. Comparison of regression tree data mining methods for prediction of mortality in head injury. Expert Syst. Appl. 2011, 38, 15534–15539. [CrossRef] Plants 2021, 10, 600 83. 84. 17 of 17 Mazaheri, H.; Ghaedi, M.; Azqhandi, M.A.; Asfaram, A. Application of machine/ statistical learning, artificial intelligence and statistical experimental design for the modeling and optimization of methylene blue and Cd (II) removal from a binary aqueous solution by natural walnut carbon. Phys. Chem. Chem. Phys. 2017, 19, 11299–11317. [CrossRef] Salonen, J.S.; Luoto, M.; Alenius, T.; Heikkilä, M.; Seppä, H.; Telford, R.J.; Birks, H.J. Reconstructing palaeoclimatic variables from fossil pollen using boosted regression trees: Comparison and synthesis with other quantitative reconstruction methods. Quat Sci. Rev. 2014, 88, 69–81. [CrossRef]

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