Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA174913

Local Sample ID:	210514_IJ3colmo_1.cdf
Subject ID:	SU001956
Subject Type:	Plant
Subject Species:	Saccharum spp

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Sample Preparation:

Sampleprep ID:	SP001962
Sampleprep Summary:	Metabolites from I5-4M and I9-4M were extracted from six biological samples of four-month-old plants. Following the removal of leaves, the internodes were identified and cut, and the bark removed (see supplementary figure S2), and the remaining tissue was immediately frozen in liquid nitrogen. Prior to metabolite extraction frozen internode tissue (100 mg) was grounded under liquid nitrogen using a vibration mill (MM 301 Retch GmbH & Co, Haan, Germany) set to a frequency of 30 Hz s-1 for 45 s, with pre-chilled holders and 3 mm tungsten beads. Internode metabolites were extracted by adding 500 μl of pre-chilled methanol (MeOH): chloroform (CHCl3): water (H20) (6:2:2 v/v/v). The extract was vortexed vigorously, sonicated at 40 Hz s-1 for 15 min and centrifuged at 14.000 x g (Eppendorf Centrifuge 5415R) for 10 min at 4°C. The supernatant was filtered (Millipore filter PVDF 0.22 μm) and 100 μl of each sample was transferred to vials and evaporated until dryness. Samples were derivatized according to Gullberg et al. (2004) with 30 μl of methoxyamine hydrochloride (15 mg ml-1) in pyridine for 16 h at room temperature. The samples were trimethylsilylated by adding 30 μl of N-methyl-N- (trimethylsilyl) trifluoroacetamide (MSTFA) containing 1% trimethylchlorosilane (TMCS), the resulting mixture stand at room temperature for 1 h. After silylation, 30 μl of heptane was added. Stable isotope reference compounds [1 mg ml-1 each of (13C3)-myristic acid, (13C4)-palmitic acid and (2H4)-succinic acid] were added in samples prior to derivatization and used as external standard for quality control. Derivatized samples were analyzed according to Gullberg et al. (2004). Blank control samples and a series of n-alkanes (C12–C40), which allowed retention indices to be calculated (Schauer et al., 2005) were also used.

Sampleprep ID:

SP001962

Sampleprep Summary:

Metabolites from I5-4M and I9-4M were extracted from six biological samples of four-month-old plants. Following the removal of leaves, the internodes were identified and cut, and the bark removed (see supplementary figure S2), and the remaining tissue was immediately frozen in liquid nitrogen. Prior to metabolite extraction frozen internode tissue (100 mg) was grounded under liquid nitrogen using a vibration mill (MM 301 Retch GmbH & Co, Haan, Germany) set to a frequency of 30 Hz s-1 for 45 s, with pre-chilled holders and 3 mm tungsten beads. Internode metabolites were extracted by adding 500 μl of pre-chilled methanol (MeOH): chloroform (CHCl3): water (H20) (6:2:2 v/v/v). The extract was vortexed vigorously, sonicated at 40 Hz s-1 for 15 min and centrifuged at 14.000 x g (Eppendorf Centrifuge 5415R) for 10 min at 4°C. The supernatant was filtered (Millipore filter PVDF 0.22 μm) and 100 μl of each sample was transferred to vials and evaporated until dryness. Samples were derivatized according to Gullberg et al. (2004) with 30 μl of methoxyamine hydrochloride (15 mg ml-1) in pyridine for 16 h at room temperature. The samples were trimethylsilylated by adding 30 μl of N-methyl-N- (trimethylsilyl) trifluoroacetamide (MSTFA) containing 1% trimethylchlorosilane (TMCS), the resulting mixture stand at room temperature for 1 h. After silylation, 30 μl of heptane was added. Stable isotope reference compounds [1 mg ml-1 each of (13C3)-myristic acid, (13C4)-palmitic acid and (2H4)-succinic acid] were added in samples prior to derivatization and used as external standard for quality control. Derivatized samples were analyzed according to Gullberg et al. (2004). Blank control samples and a series of n-alkanes (C12–C40), which allowed retention indices to be calculated (Schauer et al., 2005) were also used.