Return to study ST001214 main page
MB Sample ID: SA085839
Local Sample ID: | 4-0 |
Subject ID: | SU001281 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
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 |
---|---|---|---|---|
4-0 | SA085839 | FL012836 | lean | 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. |
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. |
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 | Triple TOF |
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: | Triple TOF |
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 |