Chn analyzer model 2400

Melting points (Mp) were determined with a Kofler hot-stage apparatus and are uncorrected. Elemental analyses were performed with a Perkin-Elmer CHN analyzer model 2400. Thin-layer chromatography: silica gel 60 F₂₅₄; layer thickness 0.2 mm (Merck, Darmstadt, Germany); eluents: a: 40% ethyl acetate/60% hexanes, b: 25% ethyl acetate/75% hexanes, detection with I₂ or UV (365 nm) after spraying with 5 % phosphomolybdic acid in 50 % aqueous phosphoric acid and heating at 100–120 °C for 10 min. Flash chromatography: silica gel 60, 40–63 μm (Merck). ¹H-NMR spectra were recorded in DMSO-d₆, CDCl₃ solution with a Bruker DRX-500 instrument at 500 MHz, with Me₄Si as internal standard. ¹³C-NMR spectra were recorded with the same instrument at 125 MHz under the same conditions. Mass spectrometry: Full scan mass spectra of the compounds were acquired in the range of 50 to 1000 m/z with a Finnigan TSQ-7000 triple quadrupole mass spectrometer (Finnigan-MAT, San Jose, CA, USA) equipped with a equipped with a Finnigan electrospray ionization source. Analyses were performed in positive ion mode using flow injection mass spectrometry with a mobile phase of 50 % aqueous acetonitrile containing 0.1 v/v % formic acid. The flow rate was 0.3 mL/min. Five µl aliquot of the samples were loaded into the flow. The ESI capillary was adjusted to 4.5 kV and N₂ was used as a nebulizer gas.

Biomass percent carbon and nitrogen composition was determined using a Perkin Elmer CHN Analyzer model 2400 (Waltham, Massachusetts, USA) per manufacturers specification. This service was completed by Analytical Service Center, Department of Forestry, University of Washington (Seattle, WA, USA).