Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA169361

Local Sample ID:	ODBS5
Subject ID:	SU001903
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	53 ± 15
Weight Or Weight Range:	116 ± 25
Gender:	Male and female

Select appropriate tab below to view additional metadata details:

Subject:

Subject ID:	SU001903
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Age Or Age Range:	53 ± 15
Weight Or Weight Range:	116 ± 25
Gender:	Male and female

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
ODBS5	SA169361	FL020202	ODBS	Genotype

Collection:

Collection ID:	CO001896
Collection Summary:	Urine, first void in the morning, were collected, aliquoted and stored at -80 °C until extraction.
Sample Type:	Urine
Collection Frequency:	First void
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001916
Treatment Summary:	150 µl of urine of each sample were randomized and vortex-mixed with 400 µl of MeOH at -20 °C containing 1 ppm of a mix of internal standards. Samples were oximated and silylated. Samples were analyzed in a Orbitrap system.

Sample Preparation:

Sampleprep ID:	SP001909
Sampleprep Summary:	30 µl of serum of each sample were randomized and vortex-mixed with 400 µl of MeOH at -20 °C containing 1 ppm of the following internal standards: heptadecanoic acid, valine-d8, succinic acid-d4 and glutamic acid-d5 (Sigma-Aldrich). Samples were incubated on ice for 30 min and centrifuged (9600 rpm, 3 min). After that, 350 µl (400 µl for urine) of the supernatant of each serum sample were transferred to a V-shaped GC-vial. Stability and reproducibility of the system were checked with pooled samples prepared colleting from all the extracts the same quantity of the remained supernatant. Afterwards, pooled samples were vortex-mixed, centrifuged and 350 µl (400 µl for urine) of the supernatant of each aliquot were transferred to a V-shaped GC-vial. Derivatization. Supernatants were evaporate to dryness in a nitrogen flow. Then, samples were converted to trimethylsilyl (TMS) and methoxime (MEOX) derivate(s). Consequently, 25 µl of MOX reagent in pyridine (20 mg/ml) were added, samples were vortex-mixed and incubated for 60 min at 45 °C. After oximation, silylation was performed adding 25 µl of MSTFA, samples were vortex-mixed and incubated for 60 min at 45 °C.

Sampleprep ID:

SP001909

Sampleprep Summary:

30 µl of serum of each sample were randomized and vortex-mixed with 400 µl of MeOH at -20 °C containing 1 ppm of the following internal standards: heptadecanoic acid, valine-d8, succinic acid-d4 and glutamic acid-d5 (Sigma-Aldrich). Samples were incubated on ice for 30 min and centrifuged (9600 rpm, 3 min). After that, 350 µl (400 µl for urine) of the supernatant of each serum sample were transferred to a V-shaped GC-vial. Stability and reproducibility of the system were checked with pooled samples prepared colleting from all the extracts the same quantity of the remained supernatant. Afterwards, pooled samples were vortex-mixed, centrifuged and 350 µl (400 µl for urine) of the supernatant of each aliquot were transferred to a V-shaped GC-vial. Derivatization. Supernatants were evaporate to dryness in a nitrogen flow. Then, samples were converted to trimethylsilyl (TMS) and methoxime (MEOX) derivate(s). Consequently, 25 µl of MOX reagent in pyridine (20 mg/ml) were added, samples were vortex-mixed and incubated for 60 min at 45 °C. After oximation, silylation was performed adding 25 µl of MSTFA, samples were vortex-mixed and incubated for 60 min at 45 °C.

Combined analysis:

Analysis ID	AN002962
Analysis type	MS
Chromatography type	Normal phase
Chromatography system	Q Exactive GC orbitrap
Column	Agilent Technologies (30 m x 0.25mm, 0.25um)
MS Type	EI
MS instrument type	Triple quadrupole
MS instrument name	Thermo Q Exactive Orbitrap
Ion Mode	POSITIVE
Units	Area

Chromatography:

Chromatography ID:	CH002195
Chromatography Summary:	GC-HRAM analysis were performed in a Q Exative GC Orbitrap system (Thermo Scientific), mounted with a Rxi Guard column purchased at Restek (10 m X 0.37 mm, 0.25 µm i.d.) and a capillary column provided by Agilent Technologies (30 m, 0.25 mm, 0.25 µm i.d.). Injection (1µ) was done in splitless mode with a TriPlus RSH autosampler system provided by Thermo. Oven temperature was keep at 70 °C for the first 5 minutes. Then, temperature was increased to 260 °C (10 °C/min) to reach in the final step 300 °C (40 °C/min) for 5 min. The carrier gas used was Helium with a flow of 2.0 ml/min. Scan range and resolution were adjusted to 50 – 500 m/z and 60,000 respectively. MS Detector was operated in EI positive mode.
Instrument Name:	Q Exactive GC orbitrap
Column Name:	Agilent Technologies (30 m x 0.25mm, 0.25um)
Chromatography Type:	Normal phase

MS:

MS ID:	MS002752
Analysis ID:	AN002962
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Triple quadrupole
MS Type:	EI
MS Comments:	GC-HRAM analysis were performed in a Q Exactive GC Orbitrap system (Thermo Scientific), mounted with a Rxi Guard column purchased at Restek (10 m X 0.37 mm, 0.25 µm i.d.) and a capillary column provided by Agilent Technologies (30 m, 0.25 mm, 0.25 µm i.d.). Injection (1µ) was done in splitless mode with a TriPlus RSH autosampler system provided by Thermo. Oven temperature was keep at 70 °C for the first 5 minutes. Then, temperature was increased to 260 °C (10 °C/min) to reach in the final step 300 °C (40 °C/min) for 5 min. The carrier gas used was Helium with a flow of 2.0 ml/min. Scan range and resolution were adjusted to 50 – 500 m/z and 60,000 respectively. MS Detector was operated in EI positive mode. The ion source and the transfer line were kept at 280 °C.
Ion Mode:	POSITIVE