Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA211387

Local Sample ID:	Fast2-2
Subject ID:	SU002292
Subject Type:	Invertebrate
Subject Species:	Caenorhabditis elegans
Taxonomy ID:	6239

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Subject:

Subject ID:	SU002292
Subject Type:	Invertebrate
Subject Species:	Caenorhabditis elegans
Taxonomy ID:	6239

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
Fast2-2	SA211387	FL025781	Fasted 8 hours	experimental factor

Collection:

Collection ID:	CO002285
Collection Summary:	C. elegans were collected after dietary restricted conditions.
Sample Type:	Worms

Treatment:

Treatment ID:	TR002304
Treatment Summary:	C. elegans were experienced restricted conditions.

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.

Combined analysis:

Analysis ID	AN003609
Analysis type	MS
Chromatography type	Unspecified
Chromatography system	Agilent 1200 RR Series II
Column	ODS
MS Type	ESI
MS instrument type	LC/MSD
MS instrument name	Agilent CE-TOFMS
Ion Mode	POSITIVE
Units	relative area

Chromatography:

Chromatography ID:	CH002668
Instrument Name:	Agilent 1200 RR Series II
Column Name:	ODS
Chromatography Type:	Unspecified

MS:

MS ID:	MS003363
Analysis ID:	AN003609
Instrument Name:	Agilent CE-TOFMS
Instrument Type:	LC/MSD
MS Type:	ESI
MS Comments:	Peaks detected in CE-TOFMS and LC-TOFMS analysis were extracted using automatic integration software (MasterHands ver. 2.17.1.11 developed at Keio University) in order to obtain peak information including m/z, migration time (MT) in CE, retention time (RT) in LC, and peak area. The peak area was then converted to relative peak area by the following equation. The peak detection limit was determined based on signal-noise ratio; S/N = 3.
Ion Mode:	POSITIVE