Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA169325

Local Sample ID:	OD10
Subject ID:	SU001902
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

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Subject:

Subject ID:	SU001902
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
OD10	SA169325	FL020198	OD	Genotype

Collection:

Collection ID:	CO001895
Collection Summary:	Serum samples were collected for CKD obese patients (OD), obese patients without CKD (O) and CKD obese patients who underwent bariatric surgery (OD BS). Samples were centrifuged (3500 rpm, 15 min at 4 °C), aliquoted and stored at -80 °C until extraction.
Sample Type:	Blood (serum)
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001915
Treatment 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 a mix of internal standards. Samples were oximated and silylated. Samples were analyzed in a Orbitrap system.

Sample Preparation:

Sampleprep ID:	SP001908
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:

SP001908

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	AN002961
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:	CH002194
Chromatography Summary:	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.
Instrument Name:	Q Exactive GC orbitrap
Column Name:	Agilent Technologies (30 m x 0.25mm, 0.25um)
Chromatography Type:	Normal phase

MS:

MS ID:	MS002751
Analysis ID:	AN002961
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