7890a 5975c gc ms

Finely milled wood powder from all transgenic (3–5 per line) and WT (33) trees was used for monosugar analysis of the greenhouse-grown trees. Extractives were removed with 80% ethanol in HEPES buffer (4 mM, pH 7.5) for 30 min at 95 °C, followed by methanol:chloroform (1:1) extraction and two washes with acetone. To remove starch, the extractive-free wood powder was treated overnight at 37 °C with α-amylase from pig pancreas (Roche 10102814001; 100 units per 100 mg of wood) in potassium phosphate buffer (0.1 M, pH 7.0). For the quantification of monosugars, 0.5 mg of amylase-treated extractive-free wood was methanolyzed using 2 M HCl/MeOH [at 85 °C for 24 h; inositol (10 μg) as internal standard] followed by trisil reagent (1,1,1,3,3,3-hexamethyldisilazane + trimethylchlorosilane + pyridine, 3:1:9) derivatization using the Sylon HTP kit (Supelco; Sigma Aldrich) [53 ]. The monosugars were separated on a J&W DB-5MS column (30 m length, 0.25 mm diameter, 0.25 μm film thickness) (Agilent Technologies) and determined using GC–MS (7890A/5975C; Agilent Technologies).

The crystal structure of the Pt-based samples was characterized by X-ray powder diffraction (XRD) using a Rigaku D/max-ga X-ray diffractometer with graphite monochromatized Cu Kα radiation (λ = 1.54178 Å). Transmission electron microscopy (TEM) images of such samples were taken using a HITACHI HT-7700 microscope operated at 100 kV. High-resolution transmission electron microscopy (HRTEM) was performed using a FEI Tecnai F30 G2 microscope operated at 300 kV. High-angle annular dark-field scanning TEM (HAADF-STEM) and Energy dispersive X-ray (EDX) mapping analyses were taken on a FEI Titan ChemiSTEM equipped with a probe-corrector and a Super-X EDX detector system and operated at 200 kV. The percentages of the elements in the samples were determined using inductively coupled plasma atomic emission spectrometry (ICP-AES, IRIS Intrepid II XSP, TJA Co., USA). Gas chromatography mass spectrometer (GC-MS) measurements were performed on a GC-MS 7890A-5975C (Agilent) with molecular ion selective monitoring. All of these samples were diluted with acetone in fixed ratio before the GC-MS measurement.

DNP−1 (7 mg) was dried overnight at 60 °C in a vacuum oven and dissolved in 5 mL DMSO and DMSO/NaOH solution (80 mg/mL, 500 μL) via sonication for 60 min. Iodomethane solution (500 μL) was added and reacted for 40 min in an ice-water bath with continuous stirring. Thereafter, 2 mL water was added to terminate the reaction.
The methylated polysaccharide was added into 2 mL TFA and hydrolyzed for 30 min at 50 °C; it was then dried via rotary evaporator at 40 °C and dissolved in 2 mL water. NaBH₄ (20 mg) was added and reacted for 30 min at 40 °C. Acetic acid was added and dried with nitrogen. Amounts of 2 mL acetic anhydride and 2mL pyridine were added and reacted for 1 h at 95 °C. Methanol was added and dried via rotary evaporator, repeated four times. Amounts of 2.5 mL water and 2.5 mL trichloromethane were added and mixed well through vortexing. The trichloromethane phase was analyzed through gas chromatography/mass (GC/MS) (GCMS-7890A/5975C, Agilent, Santa Clara, CA, USA) with an HP-5MS capillary column (initial temperature, 80 °C; increased to 280 °C at 5 °C/min; held for 1 min at 280 °C).