Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA081602

Local Sample ID:	Control 2
Subject ID:	SU001244
Subject Type:	Cultured cells
Subject Species:	Mus musculus
Taxonomy ID:	10090

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Subject:

Subject ID:	SU001244
Subject Type:	Cultured cells
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
Control 2	SA081602	FL012446	control	treatment

Collection:

Collection ID:	CO001238
Collection Summary:	Cells were washed with cold PBS and then flash-frozen in liquid N2.
Sample Type:	Muscle

Treatment:

Treatment ID:	TR001259
Treatment Summary:	Two days after confluence, cells were treated with tamoxifen for 24 h and used for the metabolomic analysis.

Sample Preparation:

Sampleprep ID:	SP001252
Sampleprep Summary:	The cells treated with vehicle (control, N = 3) or tamoxifen (FOXO1-ER, N = 3) were used for the metabolomic analysis (Human Metabolome Technologies Inc., Tsuruoka, Japan). 3.9 x 106 cells were transferred into 500 μL of methanol containing 50 mM external standard. After homogenization by BMSM10N21 (BMS, Tokyo) at 1,500 rpm for 120 s performed 5 times, 500 μL of chloroform and 200 μL of ultrapure water were added to the homogenate. The solution was mixed well and centrifuged at 2,300 × g for 5 min at 4 °C. The resultant water phase was ultrafiltered using a Millipore Ultrafree-MC PLHCC HMT Centrifugal Filter Device, 5 kDa (Millipore, Billerica, MA). The filtrate was dried and then dissolved in 50 μL of ultrapure water. The samples obtained were then subjected to capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) analysis using the Agilent CE-TOFMS system (Agilent Technologies, Santa Clara, CA) at 4 °C. The detected peaks were aligned according to their m/z values and normalized migration times. The peaks were mean-centered and scaled using their standard deviations on a per peak basis as a pretreatment.

Sampleprep ID:

SP001252

Sampleprep Summary:

The cells treated with vehicle (control, N = 3) or tamoxifen (FOXO1-ER, N = 3) were used for the metabolomic analysis (Human Metabolome Technologies Inc., Tsuruoka, Japan). 3.9 x 106 cells were transferred into 500 μL of methanol containing 50 mM external standard. After homogenization by BMSM10N21 (BMS, Tokyo) at 1,500 rpm for 120 s performed 5 times, 500 μL of chloroform and 200 μL of ultrapure water were added to the homogenate. The solution was mixed well and centrifuged at 2,300 × g for 5 min at 4 °C. The resultant water phase was ultrafiltered using a Millipore Ultrafree-MC PLHCC HMT Centrifugal Filter Device, 5 kDa (Millipore, Billerica, MA). The filtrate was dried and then dissolved in 50 μL of ultrapure water. The samples obtained were then subjected to capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) analysis using the Agilent CE-TOFMS system (Agilent Technologies, Santa Clara, CA) at 4 °C. The detected peaks were aligned according to their m/z values and normalized migration times. The peaks were mean-centered and scaled using their standard deviations on a per peak basis as a pretreatment.

Combined analysis:

Analysis ID	AN001954	AN001955
Analysis type	MS	MS
Chromatography type	None (Direct infusion)	None (Direct infusion)
Chromatography system	none	none
Column	none	none
MS Type	ESI	ESI
MS instrument type	Other	Other
MS instrument name	Agilent CE-TOFMS	Agilent CE-TOFMS
Ion Mode	POSITIVE	NEGATIVE
Units	Relative Area	fold

Chromatography:

Chromatography ID:	CH001417
Instrument Name:	none
Column Name:	none
Chromatography Type:	None (Direct infusion)

MS:

MS ID:	MS001809
Analysis ID:	AN001954
Instrument Name:	Agilent CE-TOFMS
Instrument Type:	Other
MS Type:	ESI
MS Comments:	-
Ion Mode:	POSITIVE

MS ID:	MS001810
Analysis ID:	AN001955
Instrument Name:	Agilent CE-TOFMS
Instrument Type:	Other
MS Type:	ESI
MS Comments:	-
Ion Mode:	NEGATIVE