Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA085881

Local Sample ID:	1016
Subject ID:	SU001281
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Subject:

Subject ID:	SU001281
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
1016	SA085881	FL012838	overweight	BMI group

Collection:

Collection ID:	CO001275
Collection Summary:	Blood was collected from the subjects and serum obtained by fractionation. Collection of human biomaterial, serum analyses and phenotyping were approved by the ethics committee of the University of Leipzig (approval numbers: 159-12-21052012 and 017-12-23012012), and all individuals gave written informed consent before taking part in the study.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR001296
Treatment Summary:	A cohort of 55 individuals were selected from the Leipzig biobank (42 women and 13 men) to represent a wide range of BMI (17.5–75.4 kg/m2), categories of lean (BMI < 25 kg/m2; n = 15; 4 male (M)/11 female (F)), overweight (BMI 25.1–29.9 kg/m2; n = 13; 4 M/9 F) or obese (BMI > 30 kg/m2; n = 27; 5 M/22 F) and glucose-metabolism parameters (fasting plasma glucose 3.9–13.4 mmol/liter; fasting plasma insulin 3.8–451 pmol/liter, HOMA-IR: 0.1–25). In the subgroup of lean, all individuals were normal glucose tolerant (NGT), whereas in the overweight subgroup, 10 individuals with NGT and 3 with type 2 diabetes (T2D), and in the obese group, 20 NGT individuals and 7 individuals with T2D were included.

Treatment ID:

TR001296

Treatment Summary:

A cohort of 55 individuals were selected from the Leipzig biobank (42 women and 13 men) to represent a wide range of BMI (17.5–75.4 kg/m2), categories of lean (BMI < 25 kg/m2; n = 15; 4 male (M)/11 female (F)), overweight (BMI 25.1–29.9 kg/m2; n = 13; 4 M/9 F) or obese (BMI > 30 kg/m2; n = 27; 5 M/22 F) and glucose-metabolism parameters (fasting plasma glucose 3.9–13.4 mmol/liter; fasting plasma insulin 3.8–451 pmol/liter, HOMA-IR: 0.1–25). In the subgroup of lean, all individuals were normal glucose tolerant (NGT), whereas in the overweight subgroup, 10 individuals with NGT and 3 with type 2 diabetes (T2D), and in the obese group, 20 NGT individuals and 7 individuals with T2D were included.

Sample Preparation:

Sampleprep ID:	SP001289
Sampleprep Summary:	Aliquots of 100 µL serum were taken, depending on the experiment. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at 31 −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H 2 O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H 2 O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LCMS/MS mediator lipidomics platform.

Sampleprep ID:

SP001289

Sampleprep Summary:

Aliquots of 100 µL serum were taken, depending on the experiment. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at 31 −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H 2 O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H 2 O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. (35ul) of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LCMS/MS mediator lipidomics platform.

Combined analysis:

Analysis ID	AN002025
Analysis type	MS
Chromatography type	Reversed phase
Chromatography system	Ekspert MicroLC 200 system
Column	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
MS Type	ESI
MS instrument type	QTOF
MS instrument name	ABI Sciex 5600+ TripleTOF
Ion Mode	NEGATIVE
Units	Peak Area

Chromatography:

Chromatography ID:	CH001466
Instrument Name:	Ekspert MicroLC 200 system
Column Name:	Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS001878
Analysis ID:	AN002025
Instrument Name:	ABI Sciex 5600+ TripleTOF
Instrument Type:	QTOF
MS Type:	ESI
MS Comments:	MS analysis was performed on a SCIEX TripleTOF® 5600+ system using the HR-MRM strategy consisting of a TOF MS experiment looped with multiple MS/MS experiments. MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Fullscan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX).
Ion Mode:	NEGATIVE