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Article

Phytochemical Composition of Lichen Parmotrema hypoleucinum (J. Steiner) Hale from Algeria

by
Marwa Kerboua
1,
Ali Ahmed Monia
1,
Nsevolo Samba
2,3,
Lúcia Silva
2,4,
Cesar Raposo
5,
David Díez
6 and
Jesus Miguel Rodilla
2,4,*
1
Laboratory of Vegetal Biology and Environment, Biology Department, Badji Mokhtar University, Annaba 23000, Algeria
2
Chemistry Department, University of Beira Interior, 6201-001 Covilhã, Portugal
3
Department of Clinical Analysis and Public Health, University Kimpa Vita, Uige 77, Angola
4
Fiber Materials and Environmental Technologies (FibEnTech), University of Beira Interior, 6201-001 Covilhã, Portugal
5
Mass Spectrometry Service, University of Salamanca, 37007 Salamanca, Spain
6
Department of Organic Chemistry, Faculty of Chemical Sciences, University of Salamanca, 37008 Salamanca, Spain
*
Author to whom correspondence should be addressed.
Molecules 2022, 27(16), 5229; https://doi.org/10.3390/molecules27165229
Submission received: 22 June 2022 / Revised: 18 July 2022 / Accepted: 9 August 2022 / Published: 16 August 2022
(This article belongs to the Section Natural Products Chemistry)
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Abstract

:
In this work, we carried out studies of the chemical composition of hexane, chloroform and ethanol extracts from two samples of the lichen Parmotrema hypoleucinum collected in Algeria. Each sample of the lichen P. hypoleucinum was collected on two different supports: Olea europaea and Quercus coccifera. Hexane extracts were prepared, in Soxhlet; each hexane extract was fractionated by its solubility in methanol; the products soluble in methanol were separated (cold): 1-Hexane, 2-Hexane; and the products insoluble in methanol (cold): 1-Cires, 2-Cires. A diazomethane esterified sample of 1-Hexane, 2-Hexane, 1-Cires and 2-Cires was analyzed by GC-MS, and the components were identified as methyl esters. In the 1-Hexane and 2-Hexane fractions, the methyl esters of the predominant fatty acids in the lichen were identified: palmitic acid, linoleic acid, oleic acid and stearic acid; a hydrocarbon was also identified: 13-methyl-17-norkaur-15-ene and several derivatives of orsellinic acid. In the 1-Cires and 2-Cires fractions, the previous fatty acids were no longer observed, and only the derivatives of orsellinic acid were found. The analysis of the 1-Hexane, 2-Hexane fractions by HPLC-MS/MS allows us to identify different chemical components, and the most characteristic products of the lichen were identified, such as Atranol, Chloroatranol, Atranorin and Chloroatranorin. In the fractions of 1-Cires and 2-Cires, the HPLC-MS/MS analysis reveals that they are very similar in their chemical components; the characteristic products of this lichen in this fraction are Atranorin and Chloroatranorin. In the extracts of chloroform, 1-Chloroform and 2-Chloroform, the analysis carried out by HPLC-MS/MS shows small differences in their chemical composition at the level of secondary products; among the products to be highlighted for this work, we have chloroatranorin, the stictic acid, norstictic acid and other derivatives. In the analysis of the most polar extracts carried out in ethanol: 1-Ethanol and 2-Ethanol, HPLC-MS/MS analysis shows very similar chemical compositions in these two extracts with small differences. In these extracts, the following acids were identified as characteristic compounds of this lichen: constictic acid, stictic acid, substictic acid and methylstictic acid. In the HPLC–MS/MS analysis of all these extracts, alectoronic acid was not found.

1. Introduction

Lichens live in symbiotic associations between fungi and algae and/or cyanobacteria, and in addition to these two symbiotic partners (photobiont and mycobiont) classically described, a third partner can also be integrated: epi and/or endophytic fungi as well as bacteriobiont or associated bacterial communities [1] which are important constituents of many of them. The production of various unique extracellular secondary metabolites known as lichen substances is the result of this symbiosis. The specific condition in which lichens live is the reason for the production of many metabolites that provide good protection against negative physical and biological influences. [2] The majority of lichens-forming fungi belong to Ascomycetes Lecanoromycetes [3]. Due to the vast genetic diversity and interactions with various environmental factors, lichens have unique profiles of primary and secondary metabolites (i.e., lichen substances) with interesting physiochemical properties [4].
In this paper, we describe our studies on the chemical composition of the extracts of Parmotrema hypoleucinum, Lecanorales Order, family Parmeliaceae, which, with 2765 species all over the world, is the largest family [5].
In Algeria, the Parmeliaceae family is very present compared to other families [6]; four Parmotrema have been identified so far: P. perlatum, P. reticulatum, P. robustrum and P. hypoleucinum. The last one is very common in the Mediterranean area [7]; it belongs to the Lecanorales Order and to the Parmeliaceae Family. It corresponds to a foliaceous lichen that can be up to 12 cm in diameter. In general, the lobes are very irregular, wide and raised, often forming tufts on small branches and can appear like curly lettuce. The upper side has a grayish appearance, and the lower side is largely white, which allows it to be easily distinguished from other similar species that have a dark back side. This species has black cilia and marginal Soralies.
Several activities of these lichen molecules, mainly those resulting from the polymalonate acetate pathway, are of interest for cosmetics: photoabsorbing, antioxidant and inducing melanogenesis. These properties have been studied for a still limited number of secondary metabolites (Mitrović, 2011) (Figure 1).
The natural products isolated from different lichens (such as Usnic acid, Lobaric acid, Atranorin, Protolichesterinic acid and Salazinic acid) have good antibiotic activities against Gram-positive bacteria and are also active against pathogenic dermatophyte fungi [8]. Other products found in lichens, such as anthraquinones derivatives, bianthrones and hypericin, have an inhibitory action on the activity of viral enzymes, such as the integrase of HIV-1 and HSV-1 [9,10] and also on enzymes such as lipooxygenases, histidine decarboxylase and tyrosinase; other derivatives inhibit the biosynthesis of Leukotriene B4 (LTB4) [11,12,13].
Polyphenolic products isolated from lichens have limitations, low solubility and, above all, toxicity. Usnic acid is a polyphenolic compound very common in lichens that has good activity, among others, against microorganisms of the Mycobacterium genus. In the 1950s, Shibata and Miura [14] made modifications and derivatizations of the functional groups of usnic acid to carry out the structure–activity correlation study to enhance the biological activity profile.
Usnic and polyporic acids have a good growth inhibition activity of L1210 leukemic cells; in subsequent research [15,16], several derivatives of these acids have been prepared to enhance antitumor activity, but none of these derivatives have exceeded the activities presented by usnic and polyporic acids.
Kumar and Muller have prepared a series of analogs of barbatic, diffractaic and obtusatic acids isolated from lichens to evaluate the effects of inhibition of the biosynthesis of LTB4 and as antiproliferative agents. Some of these derivatives show good potential as LTB4 biosynthesis inhibitors [11,13].
Lichens can also have xanthones, that shown enzyme modulation that are therapeutic targets, such as protein kinase C [17], topoisomerase II [18,19], acetylcholinesterase [20] and monoamine oxidases [21]; antiretrovirals [22,23], antimalarials [24,25], antihypertensives [26], anti-inflammatory, cytotoxics [27] and antitumors [28,29]. For this reason, the use of the base skeleton of xanthone is justified to prepare derivatives with bioactive potential.
Many depsidones isolated from lichens and higher plants have important activities, including the inhibition of enzymatic activity [30], antimycobacterial, anti-inflammatory, analgesic, antitumor, cytotoxic and antiviral activity [31,32,33]. In the work published in 2015 by James C Lendemer and collaborators [34], they studied and delineated the Parmotrema species in eastern North America. Using morphological, chemical, reproductive and ecological characters, they define four species for this group: P. hypoleucinum, P. hypotropum, P. perforatum and P. subrigidum.
This group has found P. hypoleucinum and P. subrigidum to be momophyletic, the latter comprising two chemotypes that differ in the presence or absence of norstictic acid in addition to alectoronic acid.
Due to the pharmacological potential presented by compounds isolated in lichens, it was decided to study the chemical composition of the lichen Parmotrema hypoleucinum (J. Steiner) Hale, collected on two different supports in the area of Lac Tonga in Algeria.

2. Results and Discussion

Parmotrema hypoleucinum (J. Steiner) Hale is an epiphytic lichen collected in Algeria and studied in order to determine its metabolic composition and chemical fingerprint. P hypoleucinum (J. Steiner) Hale was collected in two different supports, the first one in Lac Tonga (Sector Brabtia) on Olea europaea and the second one in Lac Tonga (Sector Brabtia) on Quercus coccifera. The metabolic compositions of each lichen sample were studied by sequential extraction, first of all, with hexane in Soxhlet for low polarity products. The remaining vegetable mass was placed with chloroform at room temperature to obtain the chloroform extract for the products of intermediate polarity, and finally, the vegetable mass was extracted with ethanol at room temperature for the products with higher polarity.
The hexane extract of each sample was dissolved in hot methanol and allowed to cool slowly to obtain the products insoluble in cold methanol. In this way, the products insoluble in methanol were separated: 1-Cires and 2-Cires; remaining soluble: 1-Hexane and 2-Hexane. Initially, an aliquot of the samples 1-Hexane, 2-Hexane, 1-Cires and 2-Cires were esterified with diazomethane to esterify the acid groups of the existing compounds. These esterified samples were analyzed by GC-MS to identify compounds of lower polarity.
The different extracts obtained were:
  • From P hypoleucinum (J. Steiner) Hale on Olea europaea
  • 1-Hexane, 1-Cires, 1-C 1-Ethanol
  • From Parmotrema hypoleucinum (J. Steiner) Hale on Quercus coccifera.
  • 2-Hexane, 2-Cires, 2-Choroform, 2-Ethanol
Chemical analysis of the hexane extract soluble in MeOH esterified with diazometane, 1-Hexan and 2-Hexane of Parmotrema hypoleucinum collected from two different phorophytes by GC/MS
The 1-Hexane and 2-Hexane samples were esterified with diazomethane to esterify the existing acid groups to their methyl esters for GC-MS analysis, being the natural products of the acids indicated in Table 1 for 1-Hexane sample and Table 2 for 2-Hexane sample.
The 1-Hexane sample was analyzed by GC-MS, and eight products were identified, among them palmitic, linoleic, oleic and stearic acids and an unidentified compound. Figure 2 and Table 1. The esterified 2-Hexane sample was also analyzed by GC-MS, identifying six products, palmitic, linoleic, oleic and stearic acid, a phenolic compound 2,4-dihydroxy-3,5,6-trimethylbenzoic acid and 13-methyl-17-norkaur-15-ene. Figure 3 and Table 2.
The compound identified as Hibaene (13-methyl-17-norkaur-15-ene) is the product with a retention time of 23:53 in GC-MS; its mass spectrum shows the molecular ion at 272 and comes from the dehydration of alcohol (-)-ent-Kauran-16α-ol in the ionization source of the mass spectrometer. The natural product should be the alcohol (-)-ent-Kauran-16α-ol.
P hypoleucinum (J. Steiner) Hale, on Quercus coccifera, the 2-Hexane sample, it has a lower number of components, and all were identified in the 1-Hexane sample. The most important fatty acids in the extracts were identified as palmitic, linoleic, oleic and stearic acids. 2,4-Dihydroxy-3,5,6-trimethylbenzoic acid and (-)-ent-Kauran-16α-ol alcohol is also identified in the two samples, 1-Hexane and 2-Hexane. Only in the 1-Hexane sample 4-hydroxy-2-methoxy-3,5,6-trimethylbenzoic acid and 2,4-dihydroxy-3,6-dimethylbenzoic acid are also identified.
For the fractions that have been obtained by crystallization from the crude hexane extract by solubilization in hot methanol, 1-Cires and 2-Cires are also esterified with diazomethane for GC-MS analysis.
In the 1-Cires analysis, five products are found, with four being identified. Figure 4 and Table 3.
In the 2-Cires sample, three products have been identified from the methanol-insoluble part of the hexane extract of P. hypoleucinium (Quercus coccifera). Figure 5 and Table 4.
In the 1-Cires sample, there is a compound that could not be identified; this product does not appear in the 2-Cires sample. The kaurene-skeletal alcohol is found in 1-Cires and was not found in the 2-Cires sample. Of the other three compounds, two were identified in 1-Cires and 2-Cires: 2,4-dihydroxy-3,5,6-trimetylbenzoic acid and 2,4-dihydroxy-3,6-dimethylbenzoic acid.
In the 1-Cires sample, 4-hydroxy-2-methoxy-3,6-dimethylbenzoic acid is also identified, and this product does not appear in the 2-Cires sample; instead, the 2,4-dimethoxy-6-methylbenzoic acid appears in 2-Cires.
In the HPLC—MS/MS analysis of the 1-Hexane and 2-Hexane fractions of P. hypoleucinium, 95 products were detected in the 1-Hexane fraction, in which we proposed 78 structures (and 16 not identified). In total, 91 products were found in the 2-Hexane fraction, of which we proposed 78 structures (and 13 no identified ones).
Figure 6 and Figure 7 show the HPLC chromatograms that allowed this analysis and identification of the indicated compounds to be carried out; the complete result is shown in Table 5.
The most characteristic products of the 1-Hexane and 2-Hexane extracts of the lichens in the two samples are 3-methylorsellinic acid, atranol, 4-formylbenzoic acid, chloroatranol, p-coumaric acid, atranorin and chloroatranorin.
In the fatty acid profile, we find the following acids common to the two samples: 2-hydroxy-10-undecenoic, 2-undecenedioic, undecanedioic, trans-2-dodecenedioic, 9-hydroxy-10,12-pentadecadienoic, tridecanedioic, 9Z-octadecenedioic, 10-acyl-9-formyl-13-hydroxyoctadeca-6,11-dienoic, octadecanedioic, 2-oxopalmitic, nonadecanoic, arachidic and 12-triacontenedioic acids.
Among the acids that have a cyclohexanecarboxylic base, the following derivatives were also found, as described in the publication of the lichen Physcia mediterranea Nimis [35], the 3,5-dimethoxycyclohexanecarboxylic acids, 6-(hydroxymethyl)-3,5-bis(methoxycarbonyl)-2,4-dimethylcyclohex-1-ene-1-carboxylic, 3,5,6-hydroxymethyl-2,4-dimethylcyclohex-1-ene-1-carboxylic, 5-formyl-3-hydroxymethyl-2,4,6-trimethylcyclohex-1-ene-1-carboxylic, 3,5-dihydroxy-2,4,6-trimethylcyclohex-1-ene-1-carboxylic, 5-formyl-3,6-dihydroxymethyl-2,4-dimethylcyclohex-1-ene-1-carboxylic, 2,4-dihydroxy-3,5,6-trimethylcyclohexane-1-carboxylic, 4-hydroxy-2,5-dimethylcyclohex-1-ene-1-carboxylic, 6-(1-oxopentyl)-cyclohex-1-ene-1-carboxylic acids.
Lactones were also identified: N-dodecanoyl-L-homoserine lactone and fukinanolide, other derivatives such as leoidin, lecideoidin, an alcohol such as 7,10,12-nonadecatrien-1-ol and a triterpenic acid identified as ursolic acid (found in the two extracts).
In the comparison of the characteristic polyphenolic compounds of the lichens in these two samples, 20 compounds were identified: 4-O-demethyldivaricatic, barbatic, 8-hydroxybarbatic, baeomycesic, allo-protolichesterinic, 4′-O-demethylsekikaic acids, 8-hydroxydiffractaic, glomelliferonic occurs in Xanthoparmelia subincerta [36], muronic is detected in P. praesorediosum [37], murolic, lichesterylic [35], praesorediosic, 19-acetoxyprotolichesterinic, diploicin, leprolomin, methyl 3-formyl-2-hydroxy-4-((4-methoxy-2-methylbenzoyl)oxy)-6-methylbenzoate, superpicrolichenic and 3-formyl-2-hydroxy-4-((2-(14-hydroxypentadecyl)-4-methyl-5-oxo-2,5-dihydrofuran-3-carbonyl)oxy)-6-methylbenzoic acids.
In the 1-Hexane sample, the following compounds were not identified: 3,5-dimethylorselinic acid, 9,10-dihydroxy-8-oxo-12-octadecenoic acid, octadecanedioic acid and 18-hydroxylinoleic acid, which were found in the 2-Hexane sample. The following compounds were not identified in the 2-Hexane sample: barbatic acid, tetraoxodocosanoic acid and superpicrolichenic acid; these products were identified in the 1-Hexane sample too. In the 1-Hexane sample, there are 17 products that could not be identified and in the 2-Hexane 13.
In the analysis of the 1-Cires and 2-Cires fractions, obtained by precipitation of the initial hexane extract by HPLC-MS/MS, it has been possible to detect in 1-Cires 56 products, of which 11 products were not identified. In 2-Cires, 53 products were detected, and 13 products could not be identified. For the acids and diacids identified in 1-Cires there are 23 products, and in 2-Cires, we have 27 products. As benzoic acids or derivatives we have p-coumaric acid and 6,7-dihydroxycoumarin. Among the polyphenolic compounds and esters, 1-Cires and 2-Cires were identified: allo-protolichestrinic acid, atranorin, 7-chloro-3-oxo-1,3-dihydroisobenzofuran-5-carboxylic acid, chloroatranorin, 8-hydroxydiffractaic acid and 19-acetoxylichestrinic acid.
The chromatograms and the analysis of the 1-Cires and 2-Cires compounds are shown in Figure 8 and Figure 9 and Table 6.
In the work carried out in 2016, in a general analysis of the chemical relationship in the group of Parmotrema perforatum (Parmeliaceae, Ascomycota), each sorediate species is descended from an apotheciate species with the same secondary chemicals [38]. The lichen Parmotrema hypoleucinum is derived from the ancestor Parmotrema perforatum represented by its secondary metabolites in stictic, constictic and norstictic acids. These acids have not been extracted in the Hexane extract, and they do not exist in the part insoluble in methanol, 1-Cires and 2-Cires nor in the part soluble in methanol: 1-Hexane and 2-Hexane.
The products identified in the 1-Chloroform and 2-Chloroform extracts are shown in the chromatograms in Figure 10 and Figure 11 and the results of the identification of the compounds in Table 7.
In the analysis of 1-Chloroform and 2-Chloroform extracts, the acids of the secondary metabolites that define the Sorediate species are identified for Parmotrema hypoleucinum; these acids are as follows. Constictic acid is also detected in Parmotrema tinctorum; Norstictic acid, according to mycologia 2015, this compound appeared to be present in variable concentrations throughout the thallus. Often the medulla of a lobe tested negative while the medulla adjacent to the apothecia tested positive or vice versa. Stictic acid and other derivatives identified as Substictic acid, there are depsidone were detected in Parmotrema tinctorum, P. grayanum, also in P. robustum and P. andinum [39].
In sample 1-Chloroform, 60 products were detected, of which 52 products were identified, and 8 products were not identified, and in the sample 2-Chloroform, 59 compounds were detected, of which 52 products were identified and 7 unidentified.
As in the previous analyses, fatty acids, hydroxy acids, oxo acids, some xanthones and flavones have been found, in addition to menegazziaic, siphullelic, protocetraric, conphysodalic, cryptostictic, menegazziaic isomer, gyrophoric, lecanoric and muronic acids among others.
In the extracts of the more polar products made with ethanol, in the 1-Ethanol sample 57 products were found, of which 54 products were identified, and 3 compounds were not identified. Figure 12 and Table 8. In the sample 2-Ethanol analyzed, 54 compounds were detected, of which 52 products were identified, and 2 compounds were not identified. Figure 13 and Table 8.
Among the compounds identified in the 1-Ethanol and 2-Ethanol samples were also found the acids that define this lichen Parmotrema hypoleucinum, the Constictic and Stictic acids and a derivative such as methylstictic acid.
The products analyzed by GC-MS were identified by their mass spectra and compared with the mass spectra of the NIST and Wiley databases.

3. Materials and Methods

3.1. Lichen Material

Parmotrema hypoleucinum is a foliose epiphytic lichen, which was collected on Quercus coccifera and on Olea europea at Lake Tonga (Sector Brabtia), at an altitude of 2.20 m above sea level, coordinate 36°51′38″ N; 08°28′46″ E in June 2017. The area of lake tonga is 2600 ha communicating with the sea through the artificial channel of the Messida.
This station is located in the national park of el kala (80,000 ha) Figure 14, classified as a biosphere reserve by UNESCO in 1990, located in the extreme northeast of Algeria.
Parmotrema hypoleucinum (J. Steiner) Hale was identified by Professor Monia Ali Ahmed lichenologist and research director of the Pathology of Ecosystems team at the University of Badji-Mokhtar, Annaba, Algeria. This sample has been deposited in Badji-Mokhtar University, Annaba, code AAM-2.

3.2. HPLC Orbitrap

3.2.1. Sample Preparation

Hexane extraction was carried out for each lichen sample; for Parmotrema hypoleucinium (J. Steiner), Hale was made from 40 g of powder material for the two samples. The extraction was carried out in Soxhlet apparatus with hot n-Hexane for 24 h; after this time, the solvent was evaporated to obtain the Parmotrema hypoleucinium (Olea europaea) n-Hexane extract 0.66 g, which represents (1.65%) and Parmotrema hypoleucinium (Quercus coccifera)) n-Hexane extract 0.27 g, which represent (0.68%). The n-Hexane extracts are then dissolved in hot methanol and allowed to cool to room temperature so that insoluble products crystallize. With this treatment, the methanol insoluble part (cires) and the cold methanol soluble part (Hexane) are obtained for each n-Hexane extract. For P hypoleucinium, the part insoluble in methanol produced 1-Cires of 0.379 g and 2-Cires of 0.058 g; the part soluble in methanol produced 1-Hexane of 0.281 g and 2-Hexane of 0.219 g.
The vegetable mass recovered and dried from the extractions with Hexane was placed to extract with chloroform at room temperature for 5 days. After this time, the chloroform extract was filtered, and the solvent was evaporated, obtaining the 1-Chloroform extract with a weight of 0.226 g, which represents (5.65%) and the 2-Chloroform extract with a weight of 0.196 g, which represents (4.90%).
After being extracted the vegetable mass with chloroform was placed with ethanol for 5 days to prepare the Ethanol extracts. Evaporation of the solvent gave the ethanol extracts: for 1-Ethanol, a mass of 2385 g was recovered, which represented 5.96%, and for the 2-Ethanol extract, it presented a mass of 2.417 g, which represented 6.04%.

3.2.2. Instruments

For the GCMS analysis, an Agilent MS220 mass spectrometer coupled to a 7890A GC was used.
HPLC analyses were carried out on an orbitrap Thermo q-Exactive mass spectrometer coupled to a Vanquish HPLC.

3.2.3. GCMS Parameters

The oven temperature was initially set to 50 °C, held for 5 min and then a ramp of 30 °C/min was applied up to 270 °C that was held for 5 additional mins. A VF-5 ms columns was used, with a length of 30 m, inner diameter 0.25 mm and layer width of 0.25 micron.
MS spectra were acquired in EI mode with a mass range from 50 uma to 600 uma.

3.2.4. LC Parameters

For the HPLC separation, a Kinetex XB-C18 (Phenomenex) with a particle size of 2.6 microns, 100 mm in length and a diameter of 2.1 mm was used as column. As solvent A, water with 0.1% of formic acid was used, and as solvent B, acetonitrile was chosen. The column flow war 0.200 mL/min. The following gradient was used (in Table 9):

3.2.5. MS Parameters

For the ionization electrospray in negative mode was used, with the following parameters: Electrospray voltage −3.8kV, Sheath gas 30, Aux gas 10, drying gas temperature 310 ºC. Capillary temp. 320 and S-lens value of 55.0.
The acquisition was performed in a mass range from 100 to 1000 uma, and an auto MS2 program was used with a fragmentation voltage of 30.

4. Conclusions

Due to the biological importance of lichens isolated natural products, studies of the chemical composition of two samples of the lichen Parmotrema hypoleucinum, collected on two different supports: Olea europaea and Quercus coccifera, in Algeria, were carried out. For each sample, the extracts of hot n-Hexane, Chloroform at room temperature and Ethanol at room temperature were carried out.
The n-Hexane extract of each sample is dissolved in hot methanol and allowed to cool slowly so that the products insoluble in methanol precipitate. The parts soluble in methanol are obtained by filtration, the solvent is evaporated, and they are designated as 1-Hexane and 2-Hexane fractions for each sample, respectively. The product insoluble in methanol, washed and dried, are designated as the 1-Cires and 2-Cires fractions, respectively. An aliquot sample of the fractions: 1-Hexane, 2-Hexane, 1-Cires and 2-Cires, were esterified with diazomethane to produce the methyl esters of the existing acids.
Esterified samples of 1-Hexane and 2-Hexane were analyzed by GC-MS to identify components of lower polarity. In both samples, the methyl esters of 2,4-dihydroxy-3,5,6-trimethylbenzoic acids, palmitic acid, linoleic acid, oleic acid, stearic acid and the hydrocarbon 13-methyl-17-norkaur-15-ene (Probably the natural product will be (-)-ent-kauran-16α-ol). 4-Hydroxy-2-methoxy-3,5,6-trimethylbenzoic and 2,4-dihydroxy-3,6-dimethylbenzoic acid methyl esters and a product that does not it has been possible to identify mass 288; these products were not found in the esterified sample of 2-Hexane.
The 1-Cires and 2-Cires esterified samples were analyzed by GC-MS to separate and identify the components of lower polarity. In the analysis carried out, 2,4-dihydroxy-3,5,6-trimethylbenzoic and 2,4-dihydroxy-3,6-dimethylbenzoic acids were found as existing products in the two samples. In the 2-Cires esterified sample, 2,4-dimethoxy-6-methylbenzoic acid, which does not exist in the esterified 1-Cires fraction, was also identified in this analysis. In the esterified fraction 1-Cires, the analysis also identified 4-hydroxy-2-methoxy-3,6-dimethylbenzoic acid, the hydrocarbon 13-methyl-17-norkaur-15-ene, and a product that was not identified with a mass of 312, which were not found in the esterified sample of 2-Cires.
The analysis, identification and comparison of the components of the original fractions of 1-Hexane and 2-Hexane by HPLC-MS/MS indicate that they are very similar and show almost no differences, except for some very minor components; in these fractions of low polarity, the most characteristic components P hypoleucinum will be: Atranol, Chloroatranol, Atranorin and Chloroatranorin; a triterpenic acid identified as Ursolic acid has also been found in both samples.
The analysis of the original 1-Cires and 2-Cires fractions by HPLC-MS/MS shows that they are very similar, presenting some small differences in some minor components. The predominant components identified are the fatty acids indicated in the Tables in addition to the predominant product Atranorin and Chloroatranorin, which will be characteristic for this lichen.
In the analysis carried out by HPLC-MS/MS of the original extracts of 1-Chloroform and 2-Chloroform of the two samples of P hypoleucinum and in the comparison of their components, small differences were found in some secondary compounds. In this analysis, the following acids can be considered as characteristic products of the lichen: Constictic acid, Norstictic acid (this acid was only found in the 1-Chloroform extract), Stictic acid, Substictic acid and Chloroatranorin.
The HPLC-MS/MS analysis of the original extracts of 1-Ethanol and 2-Ethanol did not suggest great differences, and most of their components were identified; the following compounds were found as more representative of the lichen: Constictic acid, Stictic acid, Substictic acid; Methylstictic acid was only identified in the extract of 1-Ethanol.
In this work, a more complete study of the components of the different extracts made from P. hypoleucinum was carried out due to the importance of the biological activities. In the most important identified products, in general, their biological activities were referred to derivatives of orsellinic acid, atranol, chloroatranol, atranorin, chloroatranorin, stictic acid and norstictic acid.
For these components, their antioxidant activities, apoptotic effects, cytotoxic, antimicrobial and antitumor activity are already known.

Author Contributions

Conceptualization, J.M.R. and A.A.M.; methodology, J.M.R., A.A.M., L.S. and D.D.; software, C.R. and M.K.; validation, L.S., D.D. and J.M.R.; formal analysis, C.R., N.S. and M.K.; investigation, A.A.M., J.M.R., L.S., N.S. and M.K.; writing-original draft preparation, J.M.R., A.A.M., L.S. and M.K.; writing-review and editing, J.M.R., A.A.M., and D.D.; funding acquisition, DGRSDT. All authors have read and agreed to the published version of the manuscript.

Funding

DGRSDT: Badji Mokhtar University, Algeira. We thank the Ministry of Higher Education and Scientific Research (MESRS) and the General Directorate of Scientific Research and Technological Development (DGRSDT) of Algeria for the financial support of this study. The authors thank to FibEnTech-UBI, which is financed by National Funds from Fundação para a Ciência e a Tecnologia (FCT) and Project UIDB/00195/2020 (financing by the FCT).

Institutional Review Board Statement

Not applicable.

Informed Consent Statement

Not applicable.

Conflicts of Interest

The authors declare no conflict of interest.

Sample Availability

Samples of the compounds are available from the authors.

References

  1. Nash, T.H. Lichen Biology; Cambridge University Press: Cambridge, UK, 2008; p. 498. [Google Scholar]
  2. Ranković, B. Lichen Secondary Metabolites: Bioactive Properties and Pharmaceutical Potential, 2nd ed.; Springer: Berlin/Heidelberg, Germany, 2019. [Google Scholar]
  3. Tehler, A.; Wedin, M. Systematics of Lichenized Fungi. In Lichen Biology, 2nd ed.; Cambridge University Press: London, UK, 2008; Chapter 17; pp. 336–352. [Google Scholar]
  4. Nguyen, K.V.; Thi Do, N.T.; Chandna, A.; Pham, C.V.; Doan, P.M.; Nguyen, A.Q.; Nguyen, C.K.T.; Larsson, M.; Escalante, S.; Olowokure, B.; et al. Antibiotic use and resistance in emerging economies: A situation analysis for Viet Nam. BMC Public Health 2013, 13, 1158. [Google Scholar] [CrossRef] [PubMed]
  5. Lücking, R.; Hodkinson, B.P.; Leavitt, S.D. The 2016 classification of lichenized fungi in the Ascomycota and Basidiomycota—Approaching one thousand genera. Bryologist 2017, 119, 361–416. [Google Scholar] [CrossRef]
  6. Amrani, S.; Seaward, M.R.D.; Sipman, H.J.M.; Feuerer, T. Lichenological exploration of Algeria II: Checklist of lichenized, lichenicolous and allied fungi. Herzogia 2018, 31, 817–892. [Google Scholar] [CrossRef]
  7. Nash, T.H.; Ryan, B.D.; Gries, C.; Bungartz, F. Lichen Flora of the Greater Sonoran Desert Region; Arizona State University Lichen Herbarium: Tempe, AZ, USA, 2002; Volume 1. [Google Scholar]
  8. Lawrey, J.D. The chemical ecology of lichen mycoparasites: A review. Can. J. Bot. 1995, 73, S1. [Google Scholar] [CrossRef]
  9. Ingólfsdóttir, K.; Chung, G.A.C.; Skúlason, V.G.; Gissurarson, S.R.; Vilhelmstóttir, M. Antimycobacterial activity of lichen metabolites in vitro. Eur. J. Pharm. Sci. 1998, 6, 141. [Google Scholar] [CrossRef]
  10. Cohen, P.A.; Hudson, J.B.; Towers, G.H.N. Antiviral activities of antraquinones, bianthrones and hypericin derivatives from lichens. Experientia 1996, 52, 180. [Google Scholar] [CrossRef]
  11. Neamati, N.; Hong, H.; Mazumder, A.; Wang, S.; Sunder, S.; Nicklaus, M.C.; Milne, G.W.A.; Proska, B.; Pommier, Y. Depsides and depsidones as inhibitors of HIV-1 integrase: Discovery of novel inhibitors through 3D database searching. J. Med. Chem. 1997, 40, 942. [Google Scholar] [CrossRef]
  12. Kumar, S.K.C.; Müller, K. Depsides as non-redox inhibitors of leukotriene B4 biosynthesis and HaCaT cell growth. 1. Novel analogues of barbatic and diffractaic acid. Eur. J. Med. Chem. 1999, 35, 1035–1042. [Google Scholar] [CrossRef]
  13. Kumar, S.K.C.; Müller, K. Lichen metabolites. 1. Inhibitory action against leukotriene B4 biosynthesis by a non-redox mechanism. J. Nat. Prod. 1999, 62, 817. [Google Scholar] [CrossRef]
  14. Kumar, S.K.C.; Müller, K. Depsides as non-redox inhibitors of leukotriene B4 biosynthesis and HaCaT cell growth. 2. Novel analogues of obtusatic acid. Eur. J. Med. Chem. 2000, 35, 405. [Google Scholar] [CrossRef]
  15. Shibata, S.; Miura, Y. Antibacterial effects of Lichen substances. 1. Comparative studies of various lichen substances. Japn. Med. J. 1948, 1, 518. [Google Scholar] [CrossRef]
  16. Cain, B.F. Potential anti-tumour agents. IV. Polyporic acid series. J. Chem. Soc. 1966, 1041–1045. [Google Scholar] [CrossRef]
  17. Takai, M.; Uehara, Y.; Beisler, J.A. Usnic acid derivatives as potential antineoplastic agents. J. Med. Chem. 1979, 22, 1380. [Google Scholar] [CrossRef] [PubMed]
  18. Saraiva, L.; Fresco, P.; Pinto, E.; Saouza, E.; Pinto, M.; Gonçalves, J. Inhibition of protein kinase C by synthetic xanthone derivatives. Bioorg. Med. Chem. 2003, 11, 1215. [Google Scholar] [CrossRef]
  19. Kwok, Y.; Zeng, Q.; Hurley, L.H. Topoisomerase II-mediated site-directed alkylation of DNA by psorospermin and its use in mapping other topoisomerase II poison binding sites. Proc. Natl. Acad. Sci. USA 1998, 95, 13531. [Google Scholar] [CrossRef] [PubMed]
  20. Woo, S.; Jung, J.; Lee, C.; Kwon, Y.; Na, Y. Synthesis of new xanthone analogues and their biological activity test-cytotoxicity, topoisomerase II inhibition, and DNA cross-linking study. Bioorg. Med. Chem. 2007, 17, 1163. [Google Scholar] [CrossRef]
  21. Piazzi, L.; Belluti, F.; Bisi, A.; Gobbi, S.; Rizzo, S.; Bartolini, M.; Andrisano, V.; Recanatini, M.; Rampa, A. Cholinesterase inhibitors: SAR and enzyme inhibitory activity of 3-[omega-(benzylmethylamino)alkoxy]xanthen-9-ones. Bioorg. Med. Chem. 2007, 15, 575. [Google Scholar] [CrossRef]
  22. Nuñez, M.B.; Maguna, F.P.; Okulik, N.B.; Castro, A.E. QSAR modeling of the MAO inhibitory activity of xanthones derivatives. Bioorg. Med. Chem. 2004, 14, 5611. [Google Scholar] [CrossRef]
  23. Ng, T.B.; Huang, B.; Fong, W.P.; Yeung, H.W. Anti-human immunodeficiency virus (anti-HIV) natural products with special emphasis on HIV reverse transcriptase inhibitors. Life Sci. 1997, 10, 933. [Google Scholar] [CrossRef]
  24. Groweiss, A.; Cardellina, I.J.H.; Boyd, M.R. HIV-Inhibitory Prenylated Santhones and Flavones from Maclura tinctoria. J. Nat. Prod. 2000, 63, 1537–1539. [Google Scholar] [CrossRef]
  25. Portela, C.; Afonso, C.M.M.; Pinto, M.M.M.; Ramos, M.J. Computational studies of new potential antimalarial compounds-Stereoelectronic complementarity with the receptor. J. Comput.-Aided Mol. Des. 2003, 17, 583. [Google Scholar] [CrossRef] [PubMed]
  26. Riscoe, M.; Kelly, J.X.; Winter, R. Xanthones as antimalarial agents: Discovery, mode of action, and optimization. Cur. Med. Chem. 2005, 12, 2539. [Google Scholar] [CrossRef] [PubMed]
  27. Wang, L.; Kang, J.; Chen, I.; Teng, C.; Lin, C. Antihypertensive and vasorelaxing activities of synthetic xanthone derivatives. Bioorg. Med. Chem. 2002, 10, 567. [Google Scholar] [CrossRef]
  28. Matsumoto, K.; Akao, Y.; Kobayashi, E.; Ohguchi, K.; Ito, T.; Tanaka, T.; Iinuma, M.; Nozawa, Y. Induction of apoptosis by xanthones from mangosteen in human leukemia cell lines. J. Nat. Prod. 2003, 66, 1124. [Google Scholar] [CrossRef]
  29. Gobbi, S.; Rampa, A.; Bisi, A.; Belluti, F.; Valenti, P.; Caputo, A.; Zampiron, A.; Carrara, M. Synthesis and antitumor activity of new derivatives of xanthen-9-one-4-acetic acid. J. Med. Chem. 2002, 45, 4931. [Google Scholar] [CrossRef]
  30. Pinto, M.M.M.; Sousa, M.E.; Nascimento, M.S.J. Xanthone derivatives: New insights in biological activities. Curr. Med. Chem. 2005, 12, 2517. [Google Scholar] [CrossRef]
  31. Hamano, K.; Kinoshitaokami, M.; Hemmi, A.; Sato, A.; Hisamoto, M.; Matsuda, K.; Yoda, K.; Haruyama, H.; Hosoya, T.; Tanzawa, K. Folipastatin, a new depsidone compound from Aspergillus unguis as an inhibitor of phospholipase A2. Taxonomy, fermentation, isolation, structure determination and biological properties. J. Antibiot. 1992, 45, 1195. [Google Scholar] [CrossRef]
  32. Permana, D.; Lajis, N.H.; Mackeen, M.M.; Ali, A.M.; Kitajima, M.; Takayama, H. Isolation and bioactivities of constituents of the roots of Garcinia atroviridis. J. Nat. Prod. 2001, 64, 976. [Google Scholar] [CrossRef]
  33. Ito, C.; Itoigawa, M.; Mishina, Y.; Tomiyasu, H.; Litaudon, M.; Cosson, J.; Mukainaka, T.; Tokuda, H.; Nishino, H.; Furukawa, H. Cancer chemopreventive agents. New depsidones from Garcinia plants. J. Nat. Prod. 2001, 64, 147. [Google Scholar] [CrossRef]
  34. Lendemer, J.C.; Allen, J.L.; Noel, N. The Parmotrema acid test: A look at species delineation in the P. perforatum group 40 y later. Mycologia 2015, 107, 1120–1129. [Google Scholar] [CrossRef]
  35. Kumar, K.; Siva, B.; Sarma, V.U.M.; Mohabe, S.; Reddy, A.M.; Boustie, J.; Tiwari, A.K.; Rao, N.R.; Babu, K.S. UPLC–MS/MS quantitative analysis and structural fragmentation study of five Parmotrema lichens from the Eastern Ghats. J. Pharm. Biomed. Anal. 2018, 156, 45–57. [Google Scholar] [CrossRef] [PubMed]
  36. Kerboua, M.; Ahmed, M.A.; Samba, N.; Aitfella-Lahlou, R.; Silva, L.; Boyero, J.F.; Raposo, C.; Rodilla, J.M.L. Phytochemical Investigation of New Algerian Lichen Species: Physcia mediterranea Nimis. Molecules 2021, 26, 1121. [Google Scholar] [CrossRef] [PubMed]
  37. Elix, J.A. A Catalogue of Standardized Chromatographic Data and Biosynthetic Relationships for Lichen Substances, 3rd ed.; The Author: Canberra, Australia, 2014. [Google Scholar]
  38. Solberg, Y. Chemical Constituent of the Lichen Species Cetraria islandica. J. Hattori Bot. Lab. 1986, 60, 391. [Google Scholar]
  39. Widhelm, T.J.; Egan, R.S.; Bertoletti, F.R.; Asztalos, M.J.; Kraichak, E.; Leavitt, S.D.; Lumbsch, H.T. Picking holes in traditional species delimitations: An integrative taxonomic reassessment of the Parmotrema perforatum group (Parmeliaceae, Ascomycota). Bot. J. Linn. Soc. 2016, 182, 868–884. [Google Scholar] [CrossRef]
  40. Torres-Benítez, A.; Rivera-Montalvo, M.; Sepúlveda, B.; Castro, O.N.; Nagles, E.; Mario, J.; Simirgiotis, M.J.; García-Beltrán, O.; Areche, C. Metabolomic Analysis of Two Parmotrema Lichens: P. robustum (Degel.) Hale and P. andinum (Mull. Arg.) Hale Using UHPLC-ESI-OT-MS-MS. Molecules 2017, 22, 1861. [Google Scholar] [CrossRef]
  41. Elix, J.A.; Wardlaw, J.H. 4-O-Methylconhypoprotocetraric acid, a new β-orcinol depsidone from the lichen Xanthoparmelia competita. Aust. J. Chem. 2000, 53, 1009–1010. [Google Scholar] [CrossRef]
  42. David, F.; Elix, J.A.; Samsudin, M.W. Two new aliphatic acids from the lichen Parmotrema praesorediosum. Aust. J. Chem. 1990, 43, 1297–1300. [Google Scholar] [CrossRef]
  43. Huynh, B.L.C.; Duong, T.H.; Do, T.M.L.; Pinnock, T.G.; Pratt, L.M.; Yamamoto, S.; Watarai, H.; Tanahashi, T.; Nguyen, K.P.P. New gamma-lactone carboxylic acids from the lichen Parmotrema praesorediosum (Nyl.) hale, parmeliaceae. Rec. Nat. Prod. 2016, 10, 332–340. [Google Scholar]
Molecules 27 05229 g001 550
Figure 1. Biosynthetic pathways of lichen secondary metabolites (Elix, 1996; Stocker wörgötter, 2008).
Figure 1. Biosynthetic pathways of lichen secondary metabolites (Elix, 1996; Stocker wörgötter, 2008).
Molecules 27 05229 g001
Molecules 27 05229 g002 550
Figure 2. Chromatogram of Hexane extract part soluble in MeOH esterified with diazomethane, 1-Hexane.
Figure 2. Chromatogram of Hexane extract part soluble in MeOH esterified with diazomethane, 1-Hexane.
Molecules 27 05229 g002
Molecules 27 05229 g003 550
Figure 3. Chromatogram of Hexane extract part soluble in MeOH esterified whit diazomethane, 2-Hexane.
Figure 3. Chromatogram of Hexane extract part soluble in MeOH esterified whit diazomethane, 2-Hexane.
Molecules 27 05229 g003
Molecules 27 05229 g004 550
Figure 4. Chromatogram of Hexane extract part insoluble in MeOH esterified whit diazomethane, 1-Cires.
Figure 4. Chromatogram of Hexane extract part insoluble in MeOH esterified whit diazomethane, 1-Cires.
Molecules 27 05229 g004
Molecules 27 05229 g005 550
Figure 5. Chromatogram of Hexane extract part insoluble in MeOH esterified whit diazometane, 2-Cires.
Figure 5. Chromatogram of Hexane extract part insoluble in MeOH esterified whit diazometane, 2-Cires.
Molecules 27 05229 g005
Molecules 27 05229 g006 550
Figure 6. Chromatogram of Hexane extract part soluble in MeOH, 1-Hexane.
Figure 6. Chromatogram of Hexane extract part soluble in MeOH, 1-Hexane.
Molecules 27 05229 g006
Molecules 27 05229 g007 550
Figure 7. Chromatogram of Hexane extract part soluble in MeOH esterified whit diazomethane, 2-Hexane.
Figure 7. Chromatogram of Hexane extract part soluble in MeOH esterified whit diazomethane, 2-Hexane.
Molecules 27 05229 g007
Molecules 27 05229 g008 550
Figure 8. Chromatogram of Hexane extract part insoluble in MeOH, 1-Cires.
Figure 8. Chromatogram of Hexane extract part insoluble in MeOH, 1-Cires.
Molecules 27 05229 g008
Molecules 27 05229 g009 550
Figure 9. Chromatogram of Hexane extract part insoluble in MeOH, 2-Cires.
Figure 9. Chromatogram of Hexane extract part insoluble in MeOH, 2-Cires.
Molecules 27 05229 g009
Molecules 27 05229 g010 550
Figure 10. Chromatogram of 1-Chloroform extract from P. hypoleucinum.
Figure 10. Chromatogram of 1-Chloroform extract from P. hypoleucinum.
Molecules 27 05229 g010
Molecules 27 05229 g011 550
Figure 11. Chromatogram of 2-Chloroform extract from P. hypoleucinum.
Figure 11. Chromatogram of 2-Chloroform extract from P. hypoleucinum.
Molecules 27 05229 g011
Molecules 27 05229 g012 550
Figure 12. Chromatogram of 1-Ethanol extract from P. hypoleucinum.
Figure 12. Chromatogram of 1-Ethanol extract from P. hypoleucinum.
Molecules 27 05229 g012
Molecules 27 05229 g013 550
Figure 13. Chromatogram of 2-Ethanol extract from P. hypoleucinum.
Figure 13. Chromatogram of 2-Ethanol extract from P. hypoleucinum.
Molecules 27 05229 g013
Molecules 27 05229 g014 550
Figure 14. Location of El Kala National Park (P.N.E.K., 2010).
Figure 14. Location of El Kala National Park (P.N.E.K., 2010).
Molecules 27 05229 g014
Table
Table 1. Sample Parmotrema hypoleucinum (in Olea europaea), Hexane extract part soluble in MeOH esterified whit diazomethane, 1-Hexane.
Table 1. Sample Parmotrema hypoleucinum (in Olea europaea), Hexane extract part soluble in MeOH esterified whit diazomethane, 1-Hexane.
NºRTIdentified ProductMass%Natural Compound, Structure
119:84Methyl 4-hydroxy-2-methoxy-3,5,6-trimethylbenzoate 2240.5 Molecules 27 05229 i001
220:00Methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate 21062.4 Molecules 27 05229 i002
320:34Methyl 2,4-dihydroxy-3,6-dimethylbenzoate1968.6 Molecules 27 05229 i003
322:36Methylhexadecanoate2705.3C16H32O2 palmitic acid
423:5313-methyl-17-norkaur-15-ene (hibaene)27210.1 Molecules 27 05229 i004
524:19Methyl 9,12-octadecadienoate 2944.2C18H32O2 linoleic acid
624:25Methyl (Z)-9-octadecenoate2962.1 C18H34O2 oleic acid
724:48Methyloctadecanoate2983.6C18H34O2 stearic acid
825:39Unidentified2880.8Unidentified
Table
Table 2. Sample Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part soluble in MeOH esterified whit diazometane, 2-Hexane.
Table 2. Sample Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part soluble in MeOH esterified whit diazometane, 2-Hexane.
NºRTIdentified ProductMass%Natural Compound, Structure
120:01Methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate 2102.5 Molecules 27 05229 i005
222:55Methylhexadecanoate2702.8C16H32O2 palmitic acid
323:5413-Methyl-17-norkaur-15-ene
Probably the natural product will be (-)-ent-Kauran-16α-ol 		27236.7 Molecules 27 05229 i006
424:19Methyl (Z,Z)-9,12-octadecadienoate 2941.2C18H32O2 linoleic acid
524:23Methyl (Z)-9-octadecenoate 2961.0C18H34O2 oleic acid
624:38Methyl octadecanoate2981.4C18H34O2 stearic acid
Table
Table 3. Sample Parmotrema hypoleucinum (in Olea europaea), Hexane extract part insoluble in MeOH esterified whit diazomethane, 1-Cires.
Table 3. Sample Parmotrema hypoleucinum (in Olea europaea), Hexane extract part insoluble in MeOH esterified whit diazomethane, 1-Cires.
NºRTIdentified ProductMass%Natural Compound, Structure
120:02Methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate 2101.1 Molecules 27 05229 i007
220:21Methyl 4-hydroxy-2-methoxy-3,6-dimethylbenzoate 210 26.6 Molecules 27 05229 i008
320:33Methyl 2,4-dihydroxy-3,6-dimethylbenzoate 196
C9H10O4		38.9 Molecules 27 05229 i009
422:51Unidentified3120.9Unidentified
522:5113-Methyl-17-nor-8β,13β-kaur-15-ene
Probably the natural product will be (-)-ent-Kauran-16α-ol 		272
C20H32
Hibaene		1.8 Molecules 27 05229 i010
Table
Table 4. Sample Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part insoluble in MeOH esterified whit diazomethane, 2-Cires.
Table 4. Sample Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part insoluble in MeOH esterified whit diazomethane, 2-Cires.
NºRTIdentified ProductMass%Natural Compound, Structure
120:24Methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate 2100.9 Molecules 27 05229 i011
220:35Methyl 2,4-dimethoxy-6-methylbenzoate 21041.4 Molecules 27 05229 i012
320:38Methyl 2,4-dihydroxy-3,6-dimethylbenzoate 19648.9 Molecules 27 05229 i013
Table
Table 5. Samples Parmotrema hypoleucinum (in Olea europea), Hexane extract part soluble in MeOH 1-Hexane and Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part soluble in MeOH, 2-Hexane.
Table 5. Samples Parmotrema hypoleucinum (in Olea europea), Hexane extract part soluble in MeOH 1-Hexane and Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part soluble in MeOH, 2-Hexane.
NºRT[M-H]−Mass CalcFormulaFormulaCompounds
1-Hexane2-Hexane
12.67181.0502182.0574C9H10O4C9H10O43-Methylorsellinic acid
22.72187.0970188.1043C9H16O4C9H16O43,5-Dimethoxyciclohexanecarboxilic acid
32.75293.0669294.0741C14H14O7C14H14O76-(Hydroxymethyl)-3,5-bis(methoxycarbonyl)-2,4-dimethylcyclohex-1-ene-1-carboxylic acid
43.08243.1239244.1311C12H20O5C12H20O53,5,6-Hydroxymethyl-2,4-dimethylcyclohex-1-ene-1-carboxylic acid
53.15151.0393152.0465C8H8O3C8H8O3Atranol
63.20225.1129226.1201C12H18O4C12H18O45-Formyl-3-hydroxymethyl-2,4,6-trimethylcyclohex-1-ene-1-carboxylic acid
73.47199.0973200.1046C10H16O4C10H16O43,5-Dihydroxy-2,4,6-trimethylciclohexenecarboxilic acid
83.55195.0660196.0730-C10H12O43,5-Dimethylorsellinic acid
93.58149.0237150.0310C8H6O3C8H6O34-Formylbenzoic acid
103.82241.1081242.1153C12H18O5C12H18O55-Formyl-3,6-dihydroxymethyl-2,4-dimethylcyclohex-1-enecarboxylic acid
114.10201.1129202.1202C10H18O4C10H18O42,4-Dihydroxy-3,5,6-trimethylcyclohexane-1-carboxylic acid
124.95199.1337200.1409C11H20O3C11H20O32-Hydroxy-10-undecenoic acid
135.00185.0006186.0079C8H7ClO3C8H7ClO3Chloroatranol
145.10213.1130214.1203C11H18O4C11H18O42-Undecenedioic acid
155.21169.0863170.0936C9H14O3C9H14O34-Hydroxy-2,5-dimethylcyclohex-1-ene-1-carboxylic acid
165.49163.0392164.0470C9H8O3C9H8O3p-Coumaric acid
175.60209.1181210.1253C12H18O3C12H18O36-(1-Oxopentyl)-1-cyclohexene-1-carboxylic acid
185.87215.1286216.1359C11H20O4C11H20O4Undecanedioic acid
196.57227.1288228.1360C12H20O4C12H20O4trans-Dodec-2-enedioic acid
207.44282.2078283.2150C16H29NO3C16H29NO3N-Dodecanoyl-L-Homoserine lactone
217.67329.2336330.2252-C18H34O59,10-Dihydroxy-8-oxo-12-octadecenoic acid
227.77209.0817210.0889C11H14O4C11H14O43,4-Dimethoxyhydrocinnamic acid
238.70373.1294374.1366C20H22O7C20H22O74-O-demethyldivaricatic acid
249.01253.1809254.1882C15H26O3C15H26O39-Hydroxy-10,12-pentadecadienoic acid
259.53243.1601244.1675C13H24O4C13H24O4Tridecanedioic acid
2610.13359.1139360.1211C19H20O7-Barbatic acid
2710.30375.1086376.1159C19H20O8C19H20O88-Hydroxybarbatic acid
2810.35373.0931374.1001C19H18O8C19H18O8Baeomycesic acid
2910.59311.2230312.2302C18H32O4C18H32O49Z-Octadecenedioic acid
3010.77351.2178352.2256C20H32O5C20H32O5 Molecules 27 05229 i014
structure proposed
3110.87233.1547234.1619C15H22O2C15H22O2Fukinanolide
3210.89323.2230324.2303C19H32O4C19H32O4allo-Protolichestrinic acid
3310.91411.0045412.0118C18H14Cl2O7C18H14Cl2O7Leoidin
3410.91396.9888397.9961C17H12Cl2O7C17H12Cl2O7Lecideoidin
3511.49403.1399404.1473C21H24O8C21H24O84′-O-demethylsekikaic acid
3612.28313.2388314.2266-C18H34O4Octadecanedioic acid
3713.00269.2124270.2195C16H30O3C16H30O32-Oxopalmitic acid
3813.04389.1245390.1315C20H22O8C20H22O88-Hydroxydiffractaic acid
3913.34293.2124294.2202C18H30O3C18H30O32-Hydroxylinolenic acid
4013.58291.1968292.2041C18H28O3C18H28O3α-Licanic acid
4114.38295.2279296.2351C18H32O3C18H32O32-Hydroxylinoleic acid isomer
4214.39455.1711456.1783C25H28O8C25H28O8Glomelliferonic acid
4314.40295.2278296.2352-C18H32O318-Hydroxylinoleic acid
4414.68321.2437322.2509C20H34O3C20H34O3Hydroxyeicosatrienoic acid
4514.95305.2125306.2197C19H30O3C19H30O314-Oxo-7,10,12-nonadecatrienoic acid
4615.19297.2436298.2508C18H34O3C18H34O39-Oxooctadecanoic acid
4715.25295.2280296.2351C18H32O3C18H32O3Coriolic acid
4815.36297.2435298.2508C18H34O3C18H34O3Ricinoleic acid
4915.42373.0925374.0999C19H18O8C19H18O8Atranorin
5015.43177.0187178.0259C9H6O4C9H6O46,7-Dihydroxycoumarin
5115.44277.2536278.2610C19H34OC19H34O7,10,12-nonadecatrien-1-ol
5215.46365.2330366.2403C21H34O5C21H34O5Muronic acid
5315.70277.2538278.2612C18H30O2C18H30O2Linolenic acid
5415.98367.2488368.2562C21H36O5C21H36O5Murolic acid
5516.16311.2594312.2667C19H36O3C19H36O3Lichesterylic acid
5616.29461.2550462.2619C26H38O7C26H38O7Unidentified
5716.37471.3481472.3553C30H48O4C30H48O4Unidentified
5816.45381.2282382.2356C21H34O6C21H34O6Praesorediosic acid
5916.47210.9834211.9873C9H5ClO4C9H5ClO4 Molecules 27 05229 i015
7-Chloro-3-oxo-1,3-dihydroisobenzofuran-5- carboxylic acid.
6016.47381.2285382.2355C21H34O6C21H34O619-Acetoxy-protolichesterinic acid
6116.51407.0540408.0611C19H17ClO8C19H17ClO8Chloroatranorin
6216.90421.9285422.9363C16H10Cl4O5C16H10Cl4O5Diploicin
6316.97201.1493202.1564C11H22O3C11H22O311-Hydroxyundecanoic acid
6416.97385.2960386.3032C22H42O5C22H42O5Unidentified
6517.59387.2544388.2616C24H36O4C24H36O4Unidentified
6617.68389.1242390.1314C20H22O8C20H22O8Leprolomin
6717.69395.2442396.2520C22H36O6-structure proposed
Molecules 27 05229 i016
6817.80357.0983358.1057C19H18O7C19H18O7structure proposed:
Molecules 27 05229 i017
6917.97449.3276450.3354C35H38O7C35H38O7Unidentified
7018.04253.2173254.2244C16H30O2C16H30O2Palmitoleic acid
7118.33241.2172242.2245C15H30O2C15H30O2Pentadecanoic acid
7218.35455.3531456.3605C30H48O3C30H48O3Ursolic acid
7318.50279.2330280.2403C18H32O2C18H32O2Linoleic acid
7418.66299.2595300.2667C18H36O3C18H36O32-Hydroxyoctadecanoic acid
7518.78279.2332280.2403C18H32O2C18H32O2Linoleic acid isomer, cis,trans
7619.03497.2548498.2626C29H38O7-Superpicrolichenic acid
7719.16255.2329256.2401C16H32O2C16H32O2Palmitic acid
7819.49281.2485282.2559C18H34O2C18H34O2Oleic acid
7919.62269.2488270.2561C17H34O2C17H34O215-Methylhexadecanoic acid
8019.89269.2488270.2561C17H34O2C17H34O2Heptadecanoic acid
8119.99483.3481484.3553C31H48O4C31H48O4Unidentified
8220.07327.2543328.2616C20H40O3C20H40O32-Hydroxyeicosanoic acid
8320.17473.2548474.2626C27H38O7C27H38O7Unidentified
8420.54283.2643284.2716C18H36O2C18H36O2Stearic acid (octadecanoic acid)
8520.74309.2801310.2875C20H38O2C20H38O2Eicosenoic acid
8621.02441.3377442.3448C29H46O3C29H46O3Unidentified
8721.14297.2801298.2873C19H38O2C19H38O2Nonadecanoic acid
8821.70311.2957312.3029C20H40O2C20H40O2(Eicosanoic acid) Arachidic acid
8922.02597.4164598.4242C37H58O6-Unidentified
9022.02669.4739670.4817C41 H66 O7-Unidentified
9122.02545.2758546.2836C30H42O9C30H42O9Structure proposed and confirmed by the ions
Molecules 27 05229 i018
9222.52579.2368580.2446C36H36O7-Unidentified
9322.66753.4057754.4135C37H62O14C37H62O14Unidentified
9422.68637.4841638.4908C41H66O5C41H66O5Unidentified
9522.69751.4779752.4857C45H38O9C45H38O9Unidentified
9623.78603.3334604.3412C37H48O7C37H48O7Unidentified
9723.81633.3798634.3876C39H54O7-Unidentified
9824.12467.4109468.4187C30H56O4-12-Triacontenedioic acid
9924.51605.3483606.3561C37H50O7C37H50O7Unidentified
Table
Table 6. Samples Parmotrema hypoleucinum (in Olea europea), Hexane extract part insoluble in MeOH, 1-Cires and Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part insoluble in MeOH, 2-Cires.
Table 6. Samples Parmotrema hypoleucinum (in Olea europea), Hexane extract part insoluble in MeOH, 1-Cires and Parmotrema hypoleucinum (in Quercus coccifera), Hexane extract part insoluble in MeOH, 2-Cires.
NºRT[M-H]−MW CalcFormula FormulaCompounds
1-Cires2-Cires
12.67146.9397147.9475C4H4O6-Dihydroxyfumaric acid
20.04190.9281191.9359C5H4O8-Methanetetracarboxylic acid
31.80116.9276117.9354C4H6O4-Butendioic acid
42.19112.9845113.9932C4H2O4C4H2O42,3-Dioxobuten-1,4-dial
52.72187.0971188.1049C9H16O4C9H16O43,5-Dimethoxyciclohexanecarboxilic acid
64.12201.1129202.1207C10H18O4C10H18O42,4-Dihydroxy-3,5,6-trimethylcyclohexane-1-carboxylic acid
75.51163.0395164.0473C9H8O3-p-Coumaric acid
85.85268.1919269.1997C12H28O6C12H28O6Unidentified
96.57227.1286228.1364-C12H20O4trans-Dodec-2-enedioic acid
107.46282.2077 283.2155C16H29NO3C16H29NO3N-Dodecanoyl-L-homoserine lactone
118.09174.9556175.9634C5H4O7-2-Hydroxy-3,4-dioxopentanedioc acid
128.15293.1762294.1840C17H26O4-gingerol
138.39323.2230324.2308-C19H32O4allo-protolichestrinic acid
149.50350.2337351.2415C17H34O7C17H34O7Xylitollaurate
1510.88233.1546234.1624-C15H22O2Fukinanolide
1611.43334.2389335.2467C17H34O6C17H34O6Unidentified
1711.75311.2231312.2309C18H32O4C18H32O49Z-octadecenedioic acid
1813.16293.2126294.2204-C18H30O32-Hydroxylinolenic acid
1913.95319.2280320.2358-C20H32O35-Hydroxyeicosatetraenoic acid
2014.37295.2280296.2358C18H32O3C18H32O318-Hydroxylinoleic acid
2114.67321.2436322.2514C20H34O3-Hydroxyeicosatrienoic acid
2214.90346.2390347.2468C18H34O6C18H34O69,10,14-trihydroxy-12-oxooctadecanoic acid
2314.98297.2434298.2512C18H34O3C18H34O39-oxooctadecanoic acid
2415.36177.0186178.0264C9H6O4C9H6O46,7-Dihydroxycoumarin
2515.44373.0929374.1007C19H18O8C19H18O8Atranorin
2616.42210.9801211.9879C9H5O4ClC9H5O4Cl Molecules 27 05229 i019
7-chloro-3-oxo-1,3-dihydroisobenzofuran-5- carboxylic acid.
2716.47407.0539408.0617C19H17O8ClC19H17O8ClChloroatranorin
2817.57277.2175278.2203-C18H30O2Octadeca-9,12,15-trienoic acid
2917.65389.1246390.1324C20H22O8-8-Hydroxydiffractaic acid,
3017.77265.1480266.1558C15H22O4C15H22O4(4E,6E,9E)-Pentadeca-4,6,9-trienedioic acid
3118.03253.2331254.2249C16H30O2C16H30O2Palmitoleic acid
3218.28402.3016403.3094-C22H42O6Unidentified
3318.34241.2173242.2251C15H30O2C15H30O2Pentadecanoic acid
3418.35455.3534456.3612C30H48O3-Oleanolic acid
3518.50279.0936280.2409C18H32O2C18H32O2Linoleic acid
3618.66489.3375490.3453-C26H50O8Unidentified
3718.86403.2645404.2723-C28H36O2Unidentified
3818.91267.2331268.2409C17H32O2C17H32O2cis-9-Heptadecenoic acid
3919.15255.2329256.2407C16H32O2C16H32O2Palmitic acid
4019.35459.3271460.3349C25H48O7C25H48O7Unidentified
4119.51281.2487282.2565C18H34O2C18H34O2Oleic acid
4219.75459.3272460.3350C25H48O7-Methyl glucose isostearate
4319.89269.2488270.2566C17H34O2C17H34O2Heptadecanoic acid
4419.99307.2645308.2723C20H36O2C20H36O211,14-Eicosadienoic acid
4520.07457.3722458.3800C27H54O5C27H54O5Unidentified
4620.18295.2645296.2723C19H36O2C19H36O210E-nonadecenoic acid
4720.54283.2643284.2721C18H36O2C18H36O2Stearic acid (Octadecanoic acid)
4820.82309.2800310.2878C20H38O2C20H38O2Eicosenoic acid
4920.86505.3326506.3404-C26H50O9Unidentified
5021.05457.3722458.3800C27H54O5-Unidentified
5121.71311.2957312.3035-C20H40O2(Eicosanoic acid) arachidic acid
5222.62297.1532298.1610C12H26O8C12H26O8Unidentified
5322.67637.4836638.4914C24H60O12N7-Unidentified
5422.77339.3268340.3346C22H44O2C22H44O2Docosanoic acid
5523.08309.1743310.1821C17H26O5C17H26O5Portentol
5623.58353.2003354.2081C19H30O6C19H30O6Unidentified
5723.81311.1689312.1767C13H30O8C13H30O8Unidentified
5823.89367.3579368.3657C24H48O2C24H48O2Lignoceric acid
5924.01397.2266398.2344C21H34O7C21H34O7Stephanol
6024.60293.1793294.1871C17H26O4C17H26O4Heptadecatrienedioic acid
6124.94325.1844326.1922C14H30O8C14H30O8Unidentified
6225.33395.3895396.3973C26H52O2-Hexacosanoic acid or cerotic acid
6325.89337.2055338.2133C19H30O5C19H30O56-Oxononadeca-8,11-dienedioic acid
6426.10339.2000340.2078C15H32O8C15H32O8Unidentified
6526.24381.2317382.2395C21H34O6C21H34O619-Acetoxylichesterinic acid
6626.75425.2581426.2659C23H38O7-Asebotoxin I
6727.53321.2106322.2184C19H30O4C19H30O4Nonadecatrienedioic Acid
6827.85304.9143305.9221Noformula-Unidentified
Table
Table 7. Samples Parmotrema hypoleucinum (in Olea europea), Chloroform extract, 1-Chloroform and Parmotrema hypoleucinum (in Quercus coccifera), Chloroform extract, 2-Chloroform.
Table 7. Samples Parmotrema hypoleucinum (in Olea europea), Chloroform extract, 1-Chloroform and Parmotrema hypoleucinum (in Quercus coccifera), Chloroform extract, 2-Chloroform.
NºRT[M-H]−Mass CalcFormulaFormulaCompounds
1-Chloroform2-Chloroform
10.98174.9557175.9635C5H4O7-2-Hydroxy-3,4-dioxopentanedioic acid
21.01145.0975146.1053-C7H14O32-Hydroxyheptanoic acid
31.05112.9845113.9923-C4H2O4Acetylenedicarboxylic acid (Squaric acid)
41.12182.9882183.9960-C7H4O6Chelidonic acid
51.14341.1091342.1169-C12H22O11Sucrose
61.52215.0097216.0175C8H8O7C8H8O7D-diacetyltartaric anhydride
71.74433.0778434.0856C20H18O11C20H18O11Avicularin
81.76403.0675404.0753C19H16O10C19H16O10Euxanthic acid
91.92417.0468418.0546C19H14O11-Shoyuflavone C
102.41433.0780434.0858C20H18O11C20H18O11Morin 3-alpha-L-arabinopyranoside
112.57401.0518402.0596C19H14O10C19H14O10Shoyuflavone B
122.64403.0672404.0750C19H16O10C19H16O10Euroxanthone B
133.15447.0934448.1012C21H20O11C21H20O11Quercitrin
143.23401.0515402.0593C19H14O10C19H14O10Constictic acid
153.38182.9882183.9960C8H5O3ClC8H5O3Cl3-Chloro-4-formylbenzoic acid
163.48187.0972188.1050C9H16O4-Azelaic acid
173.63371.0412372.0490C18H12O9-Norstictic acid
183.90313.0722314.0800-C17H14O6Cirsimaritin
193.91442.1145443.1223C18H29O8Cl2-Unidentified
203.99373.0567374.0645-C18H14O9Menegazziaic acid
214.02399.0361400.0439C19H12O10-Kynapcin-28
224.13357.0617358.0695C18H14O8C18H14O8Succinyldisalicylic acid
234.24401.0515402.0593C19H14O10-Siphulellic acid
244.27373.0568374.0646C18H14O9-Protocetraric acid
255.03417.0830418.0908C20H18O10C20H18O10Conphysodalic acid
265.09385.0568386.0646C19H14O9C19H14O9Stictic acid
275.18387.0721388.0799C19H16O9C19H16O9Cryptostictic acid
286.02385.0565386.0643C19H14O9C19H14O93,3′-Carbonylbis [6-(methoxycarbonyl)-benzoic acid]
296.14431.0984432.1062C21H20O10C21H20O10Genistein 7-glucoside (Genistin)
306.32209.0849210.0927-C11H14O4Sinapyl alcohol
317.63309.1017310.1095-C15H18O71-O-cis-cinnamoyl-β-D-glucopyranose
328.11373.0568374.0646-C18H14O9Menegazziaic acid isomer
338.67371.0408372.0486C18H12O9C18H12O9Substictic acid
349.54293.1763294.1841C17H26O4C17H26O4Nordihydrocapsiate
3510.26771.1205772.1283C38H28O18C38H28O18fucofuroeckol A hepta-acetate
3610.61426.9681427.9759-C16H12O14Unidentified
3711.11475.3278476.3356C25H48O8C25H48O8Tetrahydroxypentacosanedioic acid
3812.21345.0982346.1060C18H18O7-Isooptusatic acid (or 3′-Methylevernic acid)
3913.12265.1482266.1560C15H22O4C15H22O4EthyI 4-O-methylolivetolcarboxylate
4013.34467.0985468.1063C24H20O10C24H20O10Gyrophoric acid
4113.36317.0670318.0748C16H14O7-Lecanoric acid
4213.80503.3593504.3671C27H52O8C27H52O8Tetraglyceryl monooleate
4314.47517.3745518.3823C28H54O8-13-beta-D-glucosyloxy)docosanoic acid
4414.83359.0776360.0854-C18H16O8Ramalinaic acid
4515.40365.2334366.2412C21H34O5C21H34O5Muronic acid
4615.87265.1479266.1557C15H22O4C15H22O4Ivambrin
4716.02367.2491368.2569-C21H36O5Constipatic acid or Protoconstipatic acid
4816.28297.1532298.1610C12H26O8C12H26O8Unidentified
4917.14177.0187178.0265C9H6O4C9H6O46,7-dihydroxycoumarin
5017.37309.1743310.1821C17H26O5C17H26O5Portentol
5118.01311.1690312.1768C13H28O8C13H28O8heptahydroxytridecanol
5218.03407.0541408.0619C19H17O8ClC19H17O8ClChloroatranorin
5318.05210.9801211.9879C7H10O3Cl2C7H10O3Cl22-Methoxy-3,4-dichloro-6-methyltetrahydropyran-5-one
5418.32353.2004354.2082C19H30O6C19H30O6tetraoxononadecanoic acid
5518.36421.2265422.2343-C23H34O7Sarmentologenin
5618.85397.2268398.2346C21H34O7-Stephanol
5719.13387.2544388.2622-C24H36O4Dehydrodeoxycholic acid
5819.16441.2530442.2608C23H38O8-Asebotoxin IV
5919.32325.1846326.1924C21H26O3-Linderanolide
6019.55253.2172254.2250C16H30O2C16H30O2palmitoleic acid (9-cis-hexadecenoic acid)
6119.64255.2331256.2409C16H32O2-palmitic acid
6219.74293.1796294.1874-C17H26O4Gingerol
6319.92279.2332280.2410-C18H32O2Linoleic acid
6420.35267.2332268.241C17H32O2C17H32O22-Heptadecenoic acid
6520.51283.2645284.2723C18H36O2C18H36O2Stearic acid
6620.88281.2488282.2566C18H34O2C18H34O2Oleic acid
6722.25565.3784566.3862C32H54O8-Unidentified
6823.08679.4650680.4728C35H68O12C35H68O12Unidentified
6923.11761.5977762.6055C35H68O12C35H68O12Unidentified
7023.14395.3897396.3975-C26H52O2Hexacosanoic acid
7123.84337.2057338.2135C19H30O5C19H30O5Idebenone
7223.91367.3583368.3661C24H48O2-Lignoceric acid (tetracosanoic acid)
7324.13637.4844638.4922-C34H70O10Unidentified
7424.25639.3973640.4051C31H60O13-Unidentified
7524.44381.3741382.3819C25H50O2-Pentacosanoic acid
7624.56339.2000340.2078C15H32O8C15H32O8Heptahydroxypentadecanol
7724.60679.4650680.4728C35H68O12C35H68O12Unidentified
7824.80535.3132536.3210C26H48O11C26H48O11Unidentified
Table
Table 8. Samples Parmotrema hypoleucinum (in Olea europea), Ethanol extract, 1-Ethanol and Parmotrema hypoleucinum (in Quercus coccifera), Ethanol extract, 2-Ethanol.
Table 8. Samples Parmotrema hypoleucinum (in Olea europea), Ethanol extract, 1-Ethanol and Parmotrema hypoleucinum (in Quercus coccifera), Ethanol extract, 2-Ethanol.
NºRT[M-H]−MW Calc FormulaFormulaCompounds
1-Ethanol2-Ethanol
10.98174.9556175.9634C5H4O7C5H4O72-Hydroxy-3,4-dioxopentanedioic acid
21.05112.9845113.9923C4H2O4C4H2O4Acetylenedicarboxylic acid (Squaric acid)
31.15311.1156312.1234C15H20O7C15H20O7Neoanisatin
41.19151.0603152.0681C5H12O5C5H12O5Arabitol
51.82182.9882183.9960C7H4O6C7H4O6Chelidonic acid
62.99401.0517402.0595C19H14O10C19H14O10Constictic acid
73.95357.0617358.0695C18H14O8C18H14O8Hyposalazinic acid, Psoromic acid or Virensic acid
84.17519.1147520.1225C24H24O13-Eujambolin
94.91387.0722388.0800C19H16O9C19H16O9Cryptostictic acid
105.06385.0568386.0646C19H14O9C19H14O9Stictic acid
115.19417.0830418.0908C20H18O10C20H18O10Juglanin
125.71373.0566374.0644C18H14O9C18H14O9Protocetraric acid
135.88431.0985432.1063C21H20O10-Genistin
146.51328.0597329.0675C20H11O4N-Unidentified
156.80163.0393164.0471C9H8O3C9H8O3Coumaric acid
167.25359.0775360.0853-C18H16O8Conhypoprotocetraric acid, [40]
177.83399.0723400.0801C20H16O9-Methylstictic acid
188.44371.0408372.0486C18H12O9C18H12O9Substictic acid
198.98209.0453210.0531C10H10O5-5,6-Dihydroxy-7-methoxy-4-methyl-2-benzofuran-1(3H)-one
209.24293.1763294.1841C17H26O4C17H26O4(+)-[6]-Gingerol
219.33413.0881414.0959C21H18O9C21H18O9Vesuvianic acid
229.84461.3123462.3201-C24H46O8Unidentified
2310.28389.2911390.2989-C21H42O69,10,12,13-tetrahydroxyheneicosanoic acid
2410.49243.0065244.0143C10H9O5Cl-(4-Chloro-2-formyl-6-methoxyphenoxy)acetic Acid
2510.97475.3276476.3354C25H48O8C25H48O8Tetrahydroxypentacosanedioic acid
2611.38265.1481266.1559C15H22O4C15H22O4EthyI 4-O-methylolivetolcarboxylate
2711.42403.3068404.3146C22H44O6C22H44O69,10,12,13-Tetrahydroxydocosanoic acid
2811.62447.3331448.3409-C24H48O7D-Glucitol monostearate
2911.96489.3434490.3512C26H50O8C26H50O8Icosanedioic acid bis(2,3-dihydroxypropyl) ester
3012.05345.0982346.1060C18H18O7C18H18O7Isooptusatic acid (or 3′-Methylevernic acid)
3112.54417.3224418.33.02C23H46O6C23H46O6Heptadecyl D-glucoside
3213.20343.0823344.0901C18H16O7C18H16O7Usnic acid
3313.57431.3379432.3457C24H48O6C24H48O66-Ethyl-6-n-pentyl-pentadecan -4,5,7,8,15-pentol-I5-acetate
3413.73503.3593504.3671C27H52O8C27H52O8Tetraglyceryl monooleate
3514.25309.1746310.1824C17H26O5C17H26O5Portentol
3614.38293.2126294.2204C18H30O3C18H30O317-Hydroxylinolenic acid
3714.40517.3750518.3828C28H54O8C28H54O813-(beta-D-Glucosyloxy)docosanoic acid
3815.34365.2335366.2413C21H34O5C21H34O5Muronic acid
3916.68297.2438298.2516C18H34O3C18H34O3Ricinoleic acid
4018.00311.1691312.1769C13H28O8C13H28O8Heptahydroxytridecanol
4118.20353.2005354.2083C19H30O6C19H30O6Tetraoxononadecanoic acid
4218.71397.2267398.2345C21H34O7C21H34O7Stephanol
4318.93421.2269422.2347C23H34O7C23H34O7Sarmentologenin
4419.09441.2531442.2609C23H38O8C23H38O8Asebotoxin IV
4519.41485.2796486.2874C25H42O9C25H42O92-(7Z,10Z,13Z)-hexadecatrienoyl-3-(β-D-galactosyl)-sn-glycerol
4619.54253.2173254.2251C16H30O2C16H30O2palmitoleic acid (9-cis-hexadecenoic acid)
4719.83241.2172242.2250-C15H30O2Pentadecanoic acid
4819.93279.2332280.2410C18H32O2C18H32O2Linoleic acid
4920.03293.1795294.1873C17H26O4C17H26O4Gingerol
5020.50325.1844326.1922C21H26O3C21H26O3Linderanolide
5120.63255.2329256.2407C16H32O2C16H32O2Palmitic acid
5220.89281.2488282.2566C18H34O2C18H34O2Oleic acid
5321.18325.1846326.1924-C14H30O8Hexahydroxytetradecan-1-ol
5421.38591.2616592.2694C34H40O9-Scortechinone F
5522.01283.2645284.2723C18H36O2C18H36O2Stearic acid
5622.21761.5975762.6053C46H82O8C46H82O8Unidentified
5722.92373.0932374.1010C19H18O8-Baeomycesic acid
5822.94535.3136536.3214C26H48O11-Unidentified
5923.31311.2958312.3036C20H40O2C20H40O2Arachidic acid
6023.54337.2057338.2135C19H30O5C19H30O5Idebenone
6124.82339.2000340.2078C15H32O8C15H32O8Heptahydroxypentadecanol
6226.80367.3584368.3662C24H48O2C24H48O2Tetracosanoic acid
6327.03381.2319382.2397C21H34O6C21H34O6Praesorediosic acid or Protopraesorediosic acid [41,42,43]
Table
Table 9. Solvents used, % of solvent A and % of solvent B.
Table 9. Solvents used, % of solvent A and % of solvent B.
Time (min)%A%B
05050
200100
250100
265050
Total analysis time was set to 30 min.
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Kerboua, M.; Monia, A.A.; Samba, N.; Silva, L.; Raposo, C.; Díez, D.; Rodilla, J.M. Phytochemical Composition of Lichen Parmotrema hypoleucinum (J. Steiner) Hale from Algeria. Molecules 2022, 27, 5229. https://doi.org/10.3390/molecules27165229

AMA Style

Kerboua M, Monia AA, Samba N, Silva L, Raposo C, Díez D, Rodilla JM. Phytochemical Composition of Lichen Parmotrema hypoleucinum (J. Steiner) Hale from Algeria. Molecules. 2022; 27(16):5229. https://doi.org/10.3390/molecules27165229

Chicago/Turabian Style

Kerboua, Marwa, Ali Ahmed Monia, Nsevolo Samba, Lúcia Silva, Cesar Raposo, David Díez, and Jesus Miguel Rodilla. 2022. "Phytochemical Composition of Lichen Parmotrema hypoleucinum (J. Steiner) Hale from Algeria" Molecules 27, no. 16: 5229. https://doi.org/10.3390/molecules27165229

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