Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA211389

Local Sample ID:	Fed2
Subject ID:	SU002292
Subject Type:	Invertebrate
Subject Species:	Caenorhabditis elegans
Taxonomy ID:	6239

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Sample Preparation:

Sampleprep ID:	SP002298
Sampleprep Summary:	1. Sample Preparation for CE-TOFMS analysis The samples were mixed with 50% acetonitrile in water (v/v) containing internal standards (20 μM) as shown in Table 1 and homogenized by a homogenizer (1,500 rpm, 120 sec × 2 times), then, the same amount of 50% acetonitrile in water (v/v) were added.. The supernatant (400 μL) was then filtrated through 5-kDa cut-off filter(ULTRAFREE-MC-PLHCC, Human Metabolome Technologies, Yamagata, Japan) to remove macromolecules. The filtrate was centrifugally concentrated and resuspended in 25 μL of ultrapure water immediately before the measurement. 2. Sample Preparation for LC-TOFMS analysis The samples were mixed with 500 μL of 1% formic acid in acetonitrile (v/v) containing internal standards (10 μM), homogenized by a homogenizer (1,500 rpm, 120 sec × 2 times). The mixture was yet again homogenized after adding 167 μL of Milli-Q water and then centrifuged (2,300 x g, 4℃, 5min). After the supernatant was collected, 500 μL of 1% formic acid in acetonitrile (v/v) and 167 μL of MilliQ-water were added to the precipitation. The homogenization and centrifugation was performed as described previously, and the supernatant was mixed with previously collected one. The mixed supernatant was filtrated through 3-kDa cut-off filter (NANOCEP 3K OMEGA, PALL Corporation, Michigan, USA) to remove proteins and far filtrated through column (Hybrid SPE phospholipid 55261-U, Supelco, Bellefonte,PA, USA) to remove phospholipids. The filtrate was desiccated and resuspended in 200 μL of 50% isopropanol in Milli-Q water (v/v) immediately before the measurement.

Sampleprep ID:

SP002298

Sampleprep Summary:

1. Sample Preparation for CE-TOFMS analysis The samples were mixed with 50% acetonitrile in water (v/v) containing internal standards (20 μM) as shown in Table 1 and homogenized by a homogenizer (1,500 rpm, 120 sec × 2 times), then, the same amount of 50% acetonitrile in water (v/v) were added.. The supernatant (400 μL) was then filtrated through 5-kDa cut-off filter(ULTRAFREE-MC-PLHCC, Human Metabolome Technologies, Yamagata, Japan) to remove macromolecules. The filtrate was centrifugally concentrated and resuspended in 25 μL of ultrapure water immediately before the measurement. 2. Sample Preparation for LC-TOFMS analysis The samples were mixed with 500 μL of 1% formic acid in acetonitrile (v/v) containing internal standards (10 μM), homogenized by a homogenizer (1,500 rpm, 120 sec × 2 times). The mixture was yet again homogenized after adding 167 μL of Milli-Q water and then centrifuged (2,300 x g, 4℃, 5min). After the supernatant was collected, 500 μL of 1% formic acid in acetonitrile (v/v) and 167 μL of MilliQ-water were added to the precipitation. The homogenization and centrifugation was performed as described previously, and the supernatant was mixed with previously collected one. The mixed supernatant was filtrated through 3-kDa cut-off filter (NANOCEP 3K OMEGA, PALL Corporation, Michigan, USA) to remove proteins and far filtrated through column (Hybrid SPE phospholipid 55261-U, Supelco, Bellefonte,PA, USA) to remove phospholipids. The filtrate was desiccated and resuspended in 200 μL of 50% isopropanol in Milli-Q water (v/v) immediately before the measurement.