BRYOMOLECULES - D2.2 Report on phytochemical profile of 80 collected liverwort accessions

In this deliverable, we present the metabolomic profiles of minimum 86 liverworts accessions
(49 accessions of liverworts coming from axenic culture and about 37 coming from wild-
collected), which were delivered as part of the project BRYOMOLECULES. The table contains
species of the liverworts, additional important information, sample code and identified
compounds. Several accession of Marchantia polymorpha from each of the three subspecies
(polymorpha, ruderalis and montivagans) have been included. The taxonomic identification of
some of the species is still under verification and may be subjected to revision.
Delivered samples of liverworts were extracted with diethyl ether or/and methanol/chloroform
(2:1) or methanol. Samples of liverworts (No 1-49, 57 and 59-86) were extracted in mortar with
diethyl ether in the first step. Extracts were transferred into vials, evaporated and stored at -
20°C for further analysis. The diehtyl ether from the samples of liverworts were evaporated in
mortar in room temperature and they were re-extract in mortar with methanol/chloroform (2:1),
all of them were moved to the glass vials (20 mL) and then put on ultrasound for 20 minutes,
then they were filtered through syringe filter (17 mm, 0.22 um, PTFE hydrophobic), moved to the
vials and evaporated . Dried extracts were stored at -20°C for further analysis. Samples No 50-
56 were extracted with diethyl ether, while sample No 58 was extracted with methanol. All were
transferred into vials, evaporated and stored at -20°C for further analysis.
Highly advanced mass spectrometry techniques were applied for sample analyses. Ether
extracts were analysed both using gas chromatography coupled with electron impact ionisation
source and single Quadrupole mass analyser (GC-MS-EI-Q) and using high performance liquid
chromatography – high-resolution accurate mass – mass spectrometry with electrospray
ionisation source and Quadrupole-Time of Flight mass analyser (HPLC-HRAM-MS-ESI-QToF) in
negative and positive ionization mode. Methanol-chloroform extracts were analysed using
HPLC-HRAM-MS-ESI-QToF in negative and positive ionization mode. Moreover, samples No 58-
63 were analysed with GC-MS-EI-Q and HPLC-HRAM-MS-ESI-QToF. Identification was
performed using available spectrum libraries, standard compounds and fragmentation
patterns.
Attached table contains in the first column serial number (No) from 1 to 86 of all liverworts
accessions. The second column lists the species name, the third column contains additional
important descriptions and the fourth column includes codes of the accessions used during all
analyses. Identified compounds are presented in the fifth column.

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