identifier	taxonID	type	CVterm	format	language	title	description	additionalInformationURL	UsageTerms	rights	Owner	contributor	creator	bibliographicCitation
90640E19FFDE8D46FCF08012FBDB47D5.text	90640E19FFDE8D46FCF08012FBDB47D5.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Hieracium L.	<div><p>2.2. Qualitative and quantitative LC-MS analyses of SLs in the dry methanol extracts of flowering aerial parts of investigated  Hieracium species and statistical analysis of obtained data</p><p>The LC-MS analysis of the dry methanol extracts of flowering aerial parts of 28  Hieracium species was performed using a method optimized for the separation of water-soluble amino acid-SL conjugates. The identification of SLs in analysed extracts was done by the comparison of their retention times and MS spectra to those of isolated SLs (1–4), used as reference compounds. Their quantification in the extracts was performed by the external standard method, using the peak areas of the SIM chromatograms obtained by the monitoring of [M+H] + at m/z 540.20 (1 and 3) and at m/z 378.10 (2), and [M+NH 4] + at m/z 442.20 (4) (Figs S5.2 and 5.4). The regression equations of the calibration curves, their correlation coefficients (r 2), concentration ranges, limits of detection (LODs) and quantification (LOQs) are given in Table S2. The quantities of identified SLs in analysed dry extracts, expressed as mg/g of extract, are presented in Table 3.</p><p>Crepiside E (4) was detected in 26 species, in 21 of them it was the most abundant SL, and it was not detected only in  H. orieni A. Kern. and  H. macrodontoides (Zahn) Zahn. Its quantities broadly ranged from 0.26 mg /g in  H. pilosum Froel. extract to 15.57 mg /g in  H. blecicii Niketi ć extract. 8-Epiixerisamine A (3) and calophyllamine A (2) were present in all of the investigated species (the first was dominant in 5, and the second in one species). Their content varied from traces in the extracts of the same five species, to 11.14 mg /g in  H. guentheri-beckii Zahn extract (3), and to 5.31 mg /g in  H. anastrum (Degen &amp; Zahn) Niketi ć extract (2) (Table 3).</p><p>The only identified eudesmanolide-amino acid conjugate, calophyllamine B (1), was detected in only 14 species, where it was present in low quantities, ranging from traces in two extracts to 0.88 mg /g in  H. calophyllum extract. Calophyllamine B was dominant SL only in  H. macrodontoides extract (Table 3).</p><p>In order to compare the analysed species, the percentages of SLs relative to total detected SLs in each extract were presented (Fig. 3). To find compounds that significantly contribute to differentiation between species, principal component analysis (PCA) was performed. Results showed that all four SLs were highly correlated with variance, regardless of their average contents. Three guaianolides had the highest loadings along the first principal axis which explained 56.32% of the total variance (Fig. 4). Crepiside E (4) was positively (0.83), while 8- epiixerisamine A (3) (−0.89) and calopyllamine A (2) (−0.86) were negatively correlated. Eudesmanolide calophyllamine B (1) mostly contributed to the second axis, which explained smaller amounts of the data variation (29.80%). However, this compound was remarkably negatively correlated with the factor 2 (−0.96) rather contributing to it (77.98%). Therefore, all four SLs were more or less equally important for the variation, and they were distributed near the correlation circle, not deviating too far from orthogonality.</p><p>The majority of the species were clustered near the coordinate origin in PCA, as well as in nMDS, with relative SLs content decreasing in the following order: crepiside E (4), 8-epiixerisamine A (3), calophyllamine A (2), and calophyllamine B (1) (Figs. 4–5). The species that did not fully follow this order, containing much more 8- epiixerisamine A (3) than crepiside E (4) ( H. orieni,  H. spirocaule Niketi ć and  H. bupleuroides C. C. Gmel. s.l.) or an extremely large relative amount of crepiside E (4) ( H. mokragorae (Nägeli &amp; Peter) Nägeli &amp; Peter,  H. albopellitum (Zahn) Niketi ć and  H. tommasinianum K. Malý) (Fig. 3), were located on the opposite sides of the first axis (Figs. 4–5).  Hieracium pilosum was the only species with the highest relative content of calophyllamine A (2) (Fig. 3). Guaianolides were the dominant SLs in almost all of the investigated species. Only  H. macrodontoides, clearly separated along the second axis (Figs. 4–5), contained a remarkably high percentage of eudesmanolide calophyllamine B (1) (Fig. 3), although in a low absolute content (Table 3). Despite being detected in low concentrations in all of the investigated species, this SL could be a good chemosystematic marker for some groups, due to its absence in the basic sections  Pannosa (Zahn) Zahn,  Drepanoidea Monnier and  Naegeliana Zahn ex Szel ą g, as well as in more than a half of hybridogenous species that originated from  H. sect. Pannosa . On the contrary, this SL was almost always present in  H. sect. Villosa (Griseb.) Gremli (incl. hybridogenous species, except  H. neilreichii Beck) and Glauciformia (Freyn) Zahn –  Italica (Fr.) Av. Touv. ( H. tommasinianum group).</p><p>PCA and nMDS scores of the investigated species and the main taxonomic groups in many cases reflected differences between taxa (Figs. 4 and 5a). Their general positions were similar to those from the PCA and nMDS analyses of flavonoids and phenolic acids in investigated  Hieracium species (Milutinovi ć et al., 2018), but with evident overlaps between groups. After eliminating the scores of two hybridogenous species ( H. calophyllum and  H. durmitoricum (Rohlena &amp; Zahn) Niketi ć) in nMDS, the overlap was not particularly significant and the representatives of sections Pannosa and  Villosa (incl. hybridogenous species) were well separated along the second axis (Fig. 5b). These differences could be explained by the lack of eudesmanolide calophyllamine B (1) in section  Pannosa (except in traces in  H. paratrichum Niketi ć and  H. albopellitum) (Fig. 3). This compound was also not detected in  H. bupleuroides group (sect.  Drepanoidea) and  H. naegelianum Pan č i ć (sect. Naegeliana) (Fig. 3), and consequently these species had a similar position as the representatives of  H. gymnocephalum Griseb. ex Pant. group s.s. (Figs. 4–5).</p><p>UPGMA dendrogram of the investigated groups (Fig. 6) showed that  Glauciformia–Italica ( H. tommasinianum group), particularly influenced by the isolated position of  H. macrodontoides, was clearly separated from other sections. Section  Drepanoidea ( H. bupleuroides group) also differed from the rest of the sample. Hybridogenous groups (“gymh” and “vilh”) shared the same cluster, together with  H. naegelianum (sect. Naegeliana), which had been significantly separated from other groups in the UPGMA cluster of previously investigated phenolic compounds (Milutinovi ć et al., 2018). Inclusion of  H. gymnocephalum group in sect. Pannosa (“pan” and “wal” cluster) was also not supported.</p><p>In this work, proline-SL conjugates in general, as well as known crepiside E (4) were identified in  Hieracium genus and generally in whole subtribe  Hieraciinae for the first time. Crepiside E (4) was previously identified in only six other  Compositae taxa belonging to the genera  Crepis L. (Barda and Skaltsa, 2017),  Youngia Cass. (Lee et al., 2015; Miyase et al., 1985),  Prenanthes L. (Miyase et al., 1987),  Lapsana L. (Fontanel et al., 1999),  Ainsliaea DC (Wu et al., 2011) and  Elephantopus L. (Hisham et al., 1992).</p></div>	https://treatment.plazi.org/id/90640E19FFDE8D46FCF08012FBDB47D5	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Milutinović, Violeta;Niketić, Marjan;Krunić, Aleksej;Nikolić, Dejan;Petković, Miloš;Ušjak, Ljuboš;Petrović, Silvana	Milutinović, Violeta, Niketić, Marjan, Krunić, Aleksej, Nikolić, Dejan, Petković, Miloš, Ušjak, Ljuboš, Petrović, Silvana (2018): Sesquiterpene lactones from the methanol extracts of twenty-eight Hieracium species from the Balkan Peninsula and their chemosystematic significance. Phytochemistry 154: 19-30, DOI: 10.1016/j.phytochem.2018.06.008, URL: http://dx.doi.org/10.1016/j.phytochem.2018.06.008
90640E19FFD28D4AFFA687D5FD6742B1.text	90640E19FFD28D4AFFA687D5FD6742B1.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Hieracium calophyllum	<div><p>4.3. Preparation of MeOH extract of  H. calophyllum flowering heads and isolation of SLs</p><p>Dried and powdered flowering heads of  H. calophyllum (13 g) were macerated with CH 2 Cl 2 (1:15 w/v) at room temperature for 48 h. The extract was filtered, and the residual material dried at room temperature and re-extracted with MeOH (1:15 w/v) by bimaceration procedure (2 × 48 h). Combined MeOH extracts were evaporated under reduced pressure using a rotary evaporator (Büchi Rotavapor R- II, Flawil, Switzerland). The thus obtained dried MeOH extract (3 g) was then suspended in H 2 O (300 ml), extracted successively with CH 2 Cl 2 (300 ml), EtOAc (600 ml) and n -BuOH (600 ml), and then lyophilised. The thus obtained dry residue (0.5 g) was redissolved in H 2 O (8 mg /ml) and subjected to preparative HPLC with MS detection, on an Agilent LC/MS System, equipped with Zorbax SB-C 18 column (250 × 9.4 mm; 5 μm particle size). SLs were eluted using 0.1% (v/v) HCO 2 H/H 2 O (mobile phase A) and MeOH (mobile phase B), and selected gradient program: 10–20% B (5 min), 20–30% B (5 min), isocratic 30% B (3 min), 30-20% B (7 min), 20–50% B (4 min), 50–90% B (6 min), with the total run time of 30 min, post run time of 5 min, flow rate of 2 ml/min, and the injection volume (Vinj) of 100 μl. DAD was operating at 210 nm, 254 nm, 320 nm and 350 nm wavelengths. MSD was recording in the positive ion and full-scan mode in the range of m/ z 80–700 to acquire Total Ion Chromatogram (TIC). Optimized ion source parameters were as follows: fragmentor voltage of 200 V, nebulising drying gas flow of 9 L/min at 350 ̊C, nebulizer pressure of 40 psi and capillary voltage of 3000 V. Splitter was set at 1000:1. Time-based fractionation afforded eleven fractions, F 1–11, which were collected during 0.5 min (F 1–3), 1 min (F 4–6, 9), or 3 min (F 7, 8, 10, 11). Fraction F 4 (tR = 14.32 min) afforded compound 1 (1 mg), F 6 (tR = 15.55 min) compound 2 (1.5 mg), F 7 (tR = 16.72 min) compound 3 (3 mg) and F 9 (tR = 31.25 min) compound 4 (5 mg). Compounds 2 and 3 gradually decomposed upon one-week standing in D 2 O solution at temperature above 303 K. The mixture of 2 and its degradation products was further separated on Zorbax SB-C 18 column (50 × 9.4 mm; 5 μm particle size) affording 2a (0.5 mg). For this purpose, mobile phases: 0.1% (v/v) HCO 2 H/H 2 O (A) and MeOH (B), and fast gradient program: 20–30% B (5 min), 50–90% B (6 min), 90- 20% B (3 min), with the total run time of 15 min, flow rate of 2 ml/ min, and Vinj of 60 μl were used. The mixture of 3 and its degradation products was fractioned using the same column, mobile phases, flow rate and Vinj, with slightly different gradient program, i.e. 20–30% B (5 min), 30-20% B (5 min), 50–90% B (2 min), 90-20% B (1 min), and the total run time of 14 min, affording 4.</p><p>4.3.1. Calophyllamine B (1, 3 β -(β -glucopyranosyl)-oxy-8 α -hydroxy-13 α - (N-prolyl)-eudesma-1,4(15)-dien-5 α, 6 β, 7 α, 11 β H-12,6-olide)</p><p>White solid [α] D +106 (c 0.5, H 2 O). 1 H, 13 C, DEPT, HMBC, and NOESY spectroscopic data are presented in Table 2. HRMS m/z: 540.2456 [M + H] + (calcd 540.2439 for [M + H] + C 26 H 38 NO 11), qTOF-MS/MS (15 eV) m/z (rel. int.): 540.2456 (100), 494.2445 (22), 316.2084 (3), 128.0718 (4), 100.0759 (4).</p><p>4.3.2. Calophyllamine A (2, 3 β, 8 α -dihydroxy-13 α -(N-prolyl)-guaia- 4(15),10(14)-dien-5 α, 6 β, 7 α, 11 β H-12,6-olide)</p><p>White solid. 1 H, 13 C, HMBC, ROESY spectroscopic data are presented in Table 2. HRMS m/z: 378.1916 [M + H] + (calcd 378.1911 for [M+H] + C 20 H 28 NO 6), qTOF-MS/MS (15 eV) m/z (rel. int.): 378.1892 (100), 332.1865 (22), 128.0710 (10), 100.0729 (4).</p><p>4.3.3. 8-Epiixerisamine A (3, 3 β -(β -glucopyranosyl)-oxy-8 α -hydroxy-13 α - (N-prolyl)-guaia-4(15),10(14)-dien-5 α, 6 β, 7 α, 11 β H-12,6-olide)</p><p>White solid [α] D −3.6 (c 1.2, H 2 O). 1 H, 13 C, DEPT, ROESY and HMBC spectroscopic data are presented in Table 2. HRMS m/z: 540.2456 [M + H] + (calcd 540.2439 for [M + H] + C 26 H 38 NO 11), qTOF-MS/MS (15 eV) m/z (rel. int.): 540.2456 (100), 494.2414 (9), 378.1885 (22), 360.1806 (5), 332.1909 (5), 316.1909 (3), 128.0734 (10), 100.0886 (6).</p><p>4.3.4. Crepiside E (4, 3 β -(β -D-glucopyranosyl)-oxy-8 α -hydroxy-guaia- 4(15),10(14),11(13)-trien-5 α, 6 β, 7 α -12,6-olide)</p><p>Brownish gum. [α] D +24.3 (c 1.2, MeOH). 1 H, 13 C, COSY, NOESY,</p><p>TOCSY, HMBC spectroscopic data are presented in Table S1. HRMS m/ z: 447.1645 [M + Na] + (calcd 447.1625 for [M + Na] + C 21 H 28 O 9 Na), qTOF-MS/MS (25 eV) m/z (rel. int.): 263.1304 (29); 245.1204 (16); 227.1058 (100); 217.1268 (57); 209.1308 (14); 201.1261 (14); 199.1140 (62); 183.1078 (11); 181.104 (72); 175.0810 (5); 171.1170 (55); 143.0926 (12); 131.0869 (19).</p><p>4.3.5. Desacylcynaropicrin (2a, 3 β, 8 α -dihydroxy-guaia-4(15),10(14), 11(13)-trien-5 α, 6 β, 7 α -12,6-olide)</p><p>White solid, [α] D = +34 (c 0.5, H 2 O). 1 H NMR (500 MHz, D 2 O, TMS): δ H 6.25 (d, J = 3.4 Hz, H-13b); 6.16 (d, J = 3.0 Hz, H-13a); 5.45 (s, H-15b); 5.34 (s, H-15a); 5.16 (s, H-14b); 5.04 (s, H-14a); 4.64 (t, J = 6.2 Hz, H-3); 4.36 (t, J = 9.8 Hz, H-6); 4.06 (dd, J = 7.6, 4.7 Hz, H- 8); 3.06 (m, H-1); 2.99 (q, J = 9.9 Hz, H-5); 3.00 (t, J = 9.8 Hz, H-7); 2.72 (dd, J = 13.6, 4.9 Hz, H-9β); 2.33 (dd, J = 14.6, 3.8 Hz, H-9α); 2.22 (td, J = 13.5, 7.0 Hz, H-2β); 1.76 (td, J = 11.9, 9.2 Hz, H-2α). 13 C NMR (125 MHz, D 2 O): δ C 173.0 (C-12); 151.6 (C-4); 141.6 (C-10); 123.7 (C-13); 116.8 (C-14); 112.4 (C-15); 79.6 (C-6); 72.6 (C-3); 71.2 (C-8); 50.1 (C-7); 49.3 (C-5); 44.7 (C-1); 40.2 (C-9); 37.6 (C-2).</p></div>	https://treatment.plazi.org/id/90640E19FFD28D4AFFA687D5FD6742B1	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Milutinović, Violeta;Niketić, Marjan;Krunić, Aleksej;Nikolić, Dejan;Petković, Miloš;Ušjak, Ljuboš;Petrović, Silvana	Milutinović, Violeta, Niketić, Marjan, Krunić, Aleksej, Nikolić, Dejan, Petković, Miloš, Ušjak, Ljuboš, Petrović, Silvana (2018): Sesquiterpene lactones from the methanol extracts of twenty-eight Hieracium species from the Balkan Peninsula and their chemosystematic significance. Phytochemistry 154: 19-30, DOI: 10.1016/j.phytochem.2018.06.008, URL: http://dx.doi.org/10.1016/j.phytochem.2018.06.008
90640E19FFD18D4AFFA686DBFCA74710.text	90640E19FFD18D4AFFA686DBFCA74710.taxon	http://purl.org/dc/dcmitype/Text	http://rs.tdwg.org/ontology/voc/SPMInfoItems#GeneralDescription	text/html	en	Hieracium L.	<div><p>4.4. LC-MS analysis of dry MeOH extracts of flowering aerial parts of 28  Hieracium species</p><p>Dried and powdered flowering aerial parts of 28  Hieracium species were pre-extracted with CH 2 Cl 2 and then bimacerated with MeOH in the same manner as in the case of the flowering heads of  H. calophyllum . The number of individual plants from each species used for the extraction, the total masses of plant material, the masses of the obtained methanol extracts, and extraction yields are presented in Table 4. The obtained dried MeOH extracts were dissolved in MeOH (8.5 mg /ml), filtered through a membrane filter (0.22 μm) and subjected to qualitative and quantitative LC/SQ-MS analysis. For this purpose, Zorbax SBAq column (150 × 3.0 mm; 3.5 μm particle size, Agilent Technologies), operating at 25 ̊C, was used. The mobile phases consisted of 5 mM HCO 2 NH 4 in H 2 O (A), and MeOH (B). The gradient program was as follows: 5–10% B (8 min), 10–30% B (5 min), isocratic 30% B (2 min), 30-20% B (2 min), isocratic 20% B (2 min), 20–30% B (4 min), 30–40% B (5 min), 40–95% B (2 min), with the total run of 30 min, post run of 5 min, flow rate of 0.3 ml/min, and Vinj of 3 μl. DAD was operating at 210 and 254 nm, while MSD data were acquired in the positive ion mode: in the full-scan mode in the range of m/z 80–700, and in the Selected Ion Monitoring (SIM) mode at m/z 540.20 (1 and 3), 378.10 (2) of [M+H] +, and 442.20 (4) of [M+NH 4] +. Ion source parameters were the same as for the preparative LC-MS analysis. The MSD (in SIM mode) peak areas were used for the quantitative analysis by the external standard method. Isolated compounds (1–4) were used as the reference substances and dissolved in MeOH to obtain stock solutions (0.2 mg /ml). The calibration curves were constructed using ten different concentrations of 1–4. For each concentration, the data were recorded three times. Linearity, linear ranges, LODs and LOQs of the compounds were determined according to International Conference on Harmonization guidelines (ICH, 2005). Resulting regression equations were used to test the linearity by calculating the correlation coefficients (r 2), as well as to determine LODs and LOQs using the standard deviations of the response (σ) and the calibration curve slopes (a), in the following way:</p><p>LOD=3.3 × σ / a</p><p>LOQ =10 × σ / a</p><p>(1)</p><p>(2)</p></div>	https://treatment.plazi.org/id/90640E19FFD18D4AFFA686DBFCA74710	Public Domain	No known copyright restrictions apply. See Agosti, D., Egloff, W., 2009. Taxonomic information exchange and copyright: the Plazi approach. BMC Research Notes 2009, 2:53 for further explanation.		Plazi	Milutinović, Violeta;Niketić, Marjan;Krunić, Aleksej;Nikolić, Dejan;Petković, Miloš;Ušjak, Ljuboš;Petrović, Silvana	Milutinović, Violeta, Niketić, Marjan, Krunić, Aleksej, Nikolić, Dejan, Petković, Miloš, Ušjak, Ljuboš, Petrović, Silvana (2018): Sesquiterpene lactones from the methanol extracts of twenty-eight Hieracium species from the Balkan Peninsula and their chemosystematic significance. Phytochemistry 154: 19-30, DOI: 10.1016/j.phytochem.2018.06.008, URL: http://dx.doi.org/10.1016/j.phytochem.2018.06.008
