Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA168883

Local Sample ID:	C67B
Subject ID:	SU001891
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Subject:

Subject ID:	SU001891
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
C67B	SA168883	FL020086	CONTROL	Factor1

Collection:

Collection ID:	CO001884
Collection Summary:	On the day of the interview subjects were appointed for blood sample analysis within the next week at the primary care centre, where the interview took place, and a 30 ml blood sample were obtained at baseline. Fasting venous blood samples were drawn and immediately processed and were divided into 0.5 ml aliquots of plasma, serum, concentrated red blood cells and buffy coat, and stored in liquid nitrogen tanks at –190ºC until analysis.
Sample Type:	Blood (plasma)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001904
Treatment Summary:	Briefly, for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system.

Treatment ID:

TR001904

Treatment Summary:

Briefly, for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system.

Sample Preparation:

Sampleprep ID:	SP001897
Sampleprep Summary:	for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system.

Sampleprep ID:

SP001897

Sampleprep Summary:

for GC-MS analysis, protein precipitation was achieved by mixing 1 volume of plasma with 3 volumes of cold (-20°C) acetonitrile, followed by methoximation with O-methoxyamine hydrochloride (15 mg/mL) in pyridine, and silylation with BSTFA:TMSC (99:1). Finally, 20 ppm of tricosane in heptane was added as internal standard (IS). For CE-MS analysis, 100 µL of plasma was mixed with 100 µL of 0.2 M formic acid containing 5% acetonitrile and 0.4 mM methionine sulfone as IS. The sample was transferred to a centrifree ultracentrifugation device (Millipore Ireland Ltd.,Carrigtohill, Ireland) with a 30 kDa protein cutoff for deproteinization through centrifugation (2000 g, 4 °C, 70 min). For LC-MS analysis, 100 µL of plasma was mixed with 300 µL of a cold mixture (-20°C) of methanol:ethanol (1:1, v/v) for deproteinization. Samples were centrifuged (13,000 g, 4°C, 20 min). After centrifugation, 100 µL of the supernatant was directly injected into the system.

Combined analysis:

Analysis ID	AN002940	AN002941	AN002942	AN002943
Analysis type	MS	MS	MS	MS
Chromatography type	Reversed phase	Reversed phase	GC	Normal phase
Chromatography system	Agilent 1200	Agilent 1200	Agilent 7890A	Agilent 7100 CE
Column	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)	Agilent DB5-MS (30m x 0.25mm, 0.25um)	Agilent Technologies fused silica capillary 96cm x 50μm
MS Type	ESI	ESI	EI	ESI
MS instrument type	QTOF	QTOF	Single quadrupole	TOF
MS instrument name	Agilent 6520 QTOF	Agilent 6520 QTOF	Agilent 5975C	Agilent 6224 TOF
Ion Mode	POSITIVE	NEGATIVE	UNSPECIFIED	POSITIVE
Units	AREA	AREA		AREA

Chromatography:

Chromatography ID:	CH002177
Chromatography Summary:	The analysis of the samples was accomplished using an UHPLC system (1200 Infinity system, Agilent Technologies, Waldbronn, Germany), coupled to a 6520 QTOF MS (Agilent Technologies) with an ESI ion source. The sample injection volume was set up to 10 μL. The separation was achieved using a Discovery® HS C18 15cm x 2.1 mm, 3 µm (Supelco analytical) reverse phase column at thermostated 40 °C.
Instrument Name:	Agilent 1200
Column Name:	Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
Chromatography Type:	Reversed phase

Chromatography ID:	CH002178
Chromatography Summary:	An Agilent GC system (7890A) coupled to a- 5975C mass spectrometer (Agilent Technologies) was used to perform metabolite fingerprinting of plasma samples. Briefly, 2 μL of derivatized samples were automatically injected in split mode (ratio 1:10) through a split liner of ultra-inert deactivated glass wool from Agilent. The separation of the compounds was achieved using a pre-column (10 m J&W integrated with Agilent 122‐5532G) combined with a GC DB5-MS column (length, 30 m; internal diameter, 0.25 mm; and 0.25 μm film of 95% of dimethyl/5% diphenylpolysiloxane).
Instrument Name:	Agilent 7890A
Column Name:	Agilent DB5-MS (30m x 0.25mm, 0.25um)
Chromatography Type:	GC

Chromatography ID:	CH002179
Chromatography Summary:	The analysis was performed using a 7100 capillary electrophoresis (Agilent Technologies) coupled to a TOF MS 6224 mass spectrometer (Agilent Technologies), equipped with an ESI ion source. For the separation of metabolites, an Agilent Technologies fused silica capillary (total length, 96 cm; internal diameter, 50 μm) was used, working in normal polarity. Before each analysis, the capillary was washed for 5 min (950 mbar) with background electrolyte (BGE) (0.8 M formic acid solution in 10% methanol (v/v)). The sample injection was performed during 50 s at 50 mbar and, in order to improve the reproducibility of the analysis, the BGE was injected for 20 s at 100 mbar after the injection of each sample.
Instrument Name:	Agilent 7100 CE
Column Name:	Agilent Technologies fused silica capillary 96cm x 50μm
Chromatography Type:	Normal phase

MS:

MS ID:	MS002730
Analysis ID:	AN002940
Instrument Name:	Agilent 6520 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany)
Ion Mode:	POSITIVE

MS ID:	MS002731
Analysis ID:	AN002941
Instrument Name:	Agilent 6520 QTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany)
Ion Mode:	NEGATIVE

MS ID:	MS002732
Analysis ID:	AN002942
Instrument Name:	Agilent 5975C
Instrument Type:	Single quadrupole
MS Type:	EI
MS Comments:	Agilent MSD ChemStation software (Agilent Technologies); Agilent MassHunter Qualitative B.08.00; Agilent MassHunter Unknowns Analysis Tool 9.0.
Ion Mode:	UNSPECIFIED

MS ID:	MS002733
Analysis ID:	AN002943
Instrument Name:	Agilent 6224 TOF
Instrument Type:	TOF
MS Type:	ESI
MS Comments:	Agilent Technologies MassHunter WorkStation Data Acquisition B.08.00 software; Agilent Technologies MassHunter Profinder B.08.00 SP1 software (Waldbronn, Germany)
Ion Mode:	POSITIVE