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MB Sample ID: SA151618

Local Sample ID:root-32
Subject ID:SU001725
Subject Type:Plant
Subject Species:Hordeum vulgare L.
Taxonomy ID:4513

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Sample Preparation:

Sampleprep ID:SP001731
Sampleprep Summary:The frozen tissue was homogenized in liquid nitrogen and approximately 100 mg of powder sample was added into 2 ml vertical centrifuge tube. Then 1.4 ml of 100% methanol (pre-cooled at -20 °C) was added and vortexed thoroughly. Then the tubes were shaken for 10 min at 70 °C, followed by centrifugation at 11,000 g for 10 min. The supernatant was transferred into a new 10 ml sterile centrifuge tube, and 750 μl chloroform (−20 °C) and 1.5 ml deionized water (4 °C) were added. The mixture was vortexed and centrifuged at 2200 g for 15 min. The upper methanol/water phase was collected for metabolite analysis. A total of 150 μl collected solution was dried in a vacuum freeze dryer. The dried residue was derivatized in 40 μl of 15 mg ml-1 methoxylamine hydrochloride in pyridine for 120 min at 37 °C, followed by reaction with 70 μl of N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA) reagent for another 30 min at 37 °C. After the reaction, the reacted solution was transferred to glass vial insert in 2 ml glass bottle (Agilent, USA) for GC-MS determination. Meanwhile, one blank derivatization reaction (with reagent) was also prepared as a control.