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MB Sample ID: SA198315

Local Sample ID:	20181221_hProdh2_OE_Vector5_M62
Subject ID:	SU002169
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU002169
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
20181221_hProdh2_OE_Vector5_M62	SA198315	FL024103	Vector_OE	Genotype
20181221_hProdh2_OE_Vector5_M62	SA198315	FL024103	Control	Treatment

Collection:

Collection ID:	CO002162
Collection Summary:	cell were counted, washed twice with PBS then used for metabolite extraction.
Sample Type:	T-cells

Treatment:

Treatment ID:	TR002181
Treatment Summary:	T cells were transduced with lentivirus, then collected for metabolite extraction.

Sample Preparation:

Sampleprep ID:	SP002175
Sampleprep Summary:	After normalizing cell counts, 800 μL of 80 % (vol / vol) HPLC-grade methanol (Sigma) (precooled to -80 oC on dry ice) was added to fresh cells in a 1.5-mL microcentrifuge tube, then tubes were put on dry ice for 30 minutes. The tubes were then incubated on ice for 20 minutes and centrifuged at 15,000 x g for 15 min at 4 oC to pellet the cell debris. The metabolite-containing supernatant was transferred to a new 1.5-mL microcentrifuge tube on dry ice. Metabolite extraction was repeated with 400 μL of 80 % (vol / vol) HPLC-grade methanol. The cell lysate / methanol mixtures were dried by Speedvac at room temperature. The metabolites were dissolved again with 80 % (vol / vol) methanol, then centrifuged at 18,000 x g for 10 min to remove any particulates, and the metabolite mixtures were stored at -80 oC until LC-MS analysis.

Sampleprep ID:

SP002175

Sampleprep Summary:

After normalizing cell counts, 800 μL of 80 % (vol / vol) HPLC-grade methanol (Sigma) (precooled to -80 oC on dry ice) was added to fresh cells in a 1.5-mL microcentrifuge tube, then tubes were put on dry ice for 30 minutes. The tubes were then incubated on ice for 20 minutes and centrifuged at 15,000 x g for 15 min at 4 oC to pellet the cell debris. The metabolite-containing supernatant was transferred to a new 1.5-mL microcentrifuge tube on dry ice. Metabolite extraction was repeated with 400 μL of 80 % (vol / vol) HPLC-grade methanol. The cell lysate / methanol mixtures were dried by Speedvac at room temperature. The metabolites were dissolved again with 80 % (vol / vol) methanol, then centrifuged at 18,000 x g for 10 min to remove any particulates, and the metabolite mixtures were stored at -80 oC until LC-MS analysis.

Combined analysis:

Analysis ID	AN003402	AN003403
Analysis type	MS	MS
Chromatography type	HILIC	HILIC
Chromatography system	Agilent 6490	Agilent 6550
Column	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
MS Type	ESI	ESI
MS instrument type	Triple quadrupole	QTOF
MS instrument name	Agilent 6490 QQQ	Agilent 6550 QTOF
Ion Mode	POSITIVE	POSITIVE
Units	ppm	ppm

Chromatography:

Chromatography ID:	CH002515
Instrument Name:	Agilent 6490
Column Name:	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
Chromatography Type:	HILIC

Chromatography ID:	CH002516
Instrument Name:	Agilent 6550
Column Name:	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
Chromatography Type:	HILIC

MS:

MS ID:	MS003169
Analysis ID:	AN003402
Instrument Name:	Agilent 6490 QQQ
Instrument Type:	Triple quadrupole
MS Type:	ESI
MS Comments:	Two metabolomics strategies were adopted, i.e. untargeted metabolomics (aiming to unbiasedly detect all detectable metabolites) and targeted approaches (aiming to detect specifically defined metabolites, such as related metabolites in the proline metabolism and T cell metabolism). For untargeted metabolomics analysis, the optimized workflow consists of automated peak detection and integration, peak alignment, background noise subtraction, and multivariate data analysis. These steps were carried out for comprehensive metabolite phenotyping of the two groups using Agilent Mass Hunter Qualitative Analysis Software (Version B.07.0.0, build 7.0.7024.0) and Agilent Mass Profiler Professional (Version 14.5-Build 2772). The metabolites were first putatively identified based on accurate mass match (accurate mass ± 30 ppm error) and fragmentation pattern match. Putative structural annotation was carried out by searching the metabolite databases HMDB (http://www.hmdb.ca/) and METLIN (http:// metlin.scripps.edu) using the mass-to-charge ratio of the metabolic features.
Ion Mode:	POSITIVE

MS ID:	MS003170
Analysis ID:	AN003403
Instrument Name:	Agilent 6550 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Two metabolomics strategies were adopted, i.e. untargeted metabolomics (aiming to unbiasedly detect all detectable metabolites) and targeted approaches (aiming to detect specifically defined metabolites, such as related metabolites in the proline metabolism and T cell metabolism). For untargeted metabolomics analysis, the optimized workflow consists of automated peak detection and integration, peak alignment, background noise subtraction, and multivariate data analysis. These steps were carried out for comprehensive metabolite phenotyping of the two groups using Agilent Mass Hunter Qualitative Analysis Software (Version B.07.0.0, build 7.0.7024.0) and Agilent Mass Profiler Professional (Version 14.5-Build 2772). The metabolites were first putatively identified based on accurate mass match (accurate mass ± 30 ppm error) and fragmentation pattern match. Putative structural annotation was carried out by searching the metabolite databases HMDB (http://www.hmdb.ca/) and METLIN (http:// metlin.scripps.edu) using the mass-to-charge ratio of the metabolic features.
Ion Mode:	POSITIVE