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MB Sample ID: SA096483

Local Sample ID:OXGF14b-4-0h-1
Subject ID:SU001404
Subject Type:Plant
Subject Species:Oryza sativa Japonica Group
Taxonomy ID:39947

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Sample Preparation:

Sampleprep ID:SP001412
Sampleprep Summary:The rice panicle pre-cooled in liquid nitrogen were ground using a Mixer/mill (MM400; Retsch) with steel ball for 30 seconds at 30 HZ. Fifty milligram of rice panicle powder of each sample was extracted with a fixed volume (1 ml) of pre-cooled (−20 °C) extraction solvent (methanol: chloroform: water = 5: 2: 2) was added to homogenized tissues. After adding the extraction solvent, the vials/tubes were thoroughly vortexed for 1 min and then incubated on an orbital shaker (200 rpm) for 10 min at 4 °C followed by a 15 min sonication step. For phase separation, a volume of 500 µl of solvent (methanol: water = 1: 3), was added to each vial/tube and the samples were again thoroughly vortexed for 1 min. After that, the samples are centrifuged at a speed of 14000rpm for 10 min at 4 °C. Two fixed volume of 200 μL of the polar phase (the lower phase) were transferred into pre-labeled 1.5 ml microcentrifuge tube respectively. Then the samples were dried in a SpeedVac concentrator without heating. Two dried 200 μL aliquots of the polar phase in each sample were analyzed using gas chromatography tandem mass spectrometry (GC-MS) and liquid chromatography tandem mass spectrometry (LC-MS) for metabolomics study. The dried 200 µl aliquots of the polar phase for GC-MS analysis were re-suspended in methoxyamine-hydrochloride/pyridine solution for methoxymization of carbonyl groups followed by heating at 37 °C for 2 h. The samples were further derivatized with N-methyl-N-trimethylsilyltrifloracetamide (MSTFA) for 30 min at 37 °C. then one µl of the derivatized sample mixture was injected onto the GC-column and measured. Another dried 200 µl aliquots of the polar phase were re-suspended in 150 µl UPLC-grade methanol: water (1:1, vol/vol) and subjected to LC-MS analysis.
Processing Storage Conditions:-80℃
Extract Storage:-80℃
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