ABSTRACT: Plant metabolic extracts. Plant tissues as specified in file name were collected from adult live plants in the University of Michigan Matthaei Botanical Garden. 0.2g fresh weight tissue were frozen, ground in a Tissuelyzer for 2min at 6m/s with 10 2.3mm silica beads in 2mL vials. Ground tissue was extracted with 3mL methanol for 1h at 37 celsius shaking. Methanol was separated from insoluble material and dried under nitrogen. Dried crude methanol extracts were resuspended in water, partitioned twice with 3 mL hexane, 3 mL ethyl acetate and finally extracted with 3 mL n-butanol. Ethylacetate and n-butanol extracts were dried in a speedvac concentrator, resuspended in 1 mL methanol, filtered through a 0.2 micron filter and analyzed by LC-MS/MS on an Thermo QExactive orbitrap with a ESI source. LC-MS settings were as follows: Injection volume 5 uL, LC - Phenomenex Kinetex 2.6 um C18 reverse phase 100 A 150 x 3 mm LC column, LC gradient: solvent A - 0.1% formic acid, solvent B - acetonitrile (0.1% formic acid), 0 min: 10% B, 5 min: min: 60% B, 5.1 min: 95% B, 6 min: 95% B, 6.1 min: 10% B, 9.9 min: 10% B, 0.5 mL/min, MS - positive ion mode, Full MS: Resolution 70000, mass range 400-1200 m/z, dd-MS2 (data-dependent MS/MS): resolution 17500, AGC target 1e5, Loop count 5, Isolation width 1.0 m/z, Collision energy 25 eV, dynamic exclusion 0.5 s. [doi:10.25345/C5H41JX71] [dataset license: CC0 1.0 Universal (CC0 1.0)]
Desnor Chigumba