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MB Sample ID: SA162941
| Local Sample ID: | VAT (ob)3 |
| Subject ID: | SU001815 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001815 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| VAT (ob)3 | SA162941 | FL019377 | Visceral | Depot location |
| VAT (ob)3 | SA162941 | FL019377 | obese | Phenotype |
Collection:
| Collection ID: | CO001808 |
| Collection Summary: | Samples of human white adipose tissue from a total of 86 donors were kindly provided by Matthias Blüher as a part of Leipzig Obesity BioBank. Tissue collection was approved by the Ethics committee of the University of Leipzig (approval number: 159-12-21052012) and all subjects gave written informed consent before taking part in the study. Removed tissue samples were flash frozen in liquid nitrogen and stored at -80°C until further analysis. For the purpose of this study, we included adipose tissue samples from abdominal visceral (VAT) and subcutaneous (SAT) fat depots of lean (BMI < 25kg/m²; n = 5) and obese (BMI > 40kg/m²; n = 81) individuals. Representative tissue pools were generated according to depot and phenotype specificity. |
| Sample Type: | Adipose tissue |
| Collection Location: | Leipzig, Germany |
| Storage Conditions: | -80℃ |
| Storage Vials: | Plastic tubes |
| Additives: | none |
Treatment:
| Treatment ID: | TR001828 |
| Treatment Summary: | Adipose tissue biopsy pools were not treated prior to sample preparation. |
Sample Preparation:
| Sampleprep ID: | SP001821 |
| Sampleprep Summary: | Adipose tissue was spiked with internal standards, homogenized by high-speed homogenization and lipids were extracted using the Folch method. Triacylglycerols were depleted by EtOH/Hexane based liquid/liquid extraction. |
| Sampleprep Protocol Filename: | Sample Preparation Workflow |
| Processing Storage Conditions: | 4℃ |
| Extraction Method: | Folch lipid extraction |
| Extract Enrichment: | EtOH/Hexane liquid/liquid extraction |
| Extract Storage: | -80℃ |
| Sample Derivatization: | none |
| Sample Spiking: | adipose tissue specific internal standard mix |
Combined analysis:
| Analysis ID | AN002829 | AN002830 | AN002831 |
|---|---|---|---|
| Analysis type | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | HILIC |
| Chromatography system | Vanquish Horizon | Vanquish Horizon | Vanquish Horizon |
| Column | Thermo Accucore C30 (150 x 2.1mm,2.6um) | Thermo Accucore C18 (150 x 2.1mm,2.6um) | Waters ACQUITY UPLC BEH HILIC (100 x 1.0mm,1.7um,130 Å) |
| MS Type | ESI | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap; Thermo Fusion Orbitrap | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | UNSPECIFIED | UNSPECIFIED | POSITIVE |
| Units | fmol/μg protein | fmol/μg protein | fmol/μg protein |
Chromatography:
| Chromatography ID: | CH002092 |
| Chromatography Summary: | Three chromatography types were employed to allow for optimal lipidome separation (i.e. C18 RPC for amphiphilic lipids, C30 RPC for triacylglycerols, HILIC for acylcarnitines). All details on the respective methods are provided in the attached methods document |
| Instrument Name: | Vanquish Horizon |
| Column Name: | Thermo Accucore C30 (150 x 2.1mm,2.6um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0-5 min - 50 to 80 % B (curve 5), 5-22 min - 80 to 95 % B (curve 4), 22-26 min - 95 % isocratic, 26-27 min - 95 to 100 % B (curve 5), 27-47 min - 100 % B isocratic, 47-47.1 min - 100 to 50 % B followed by 8 min re-equilibration at 50% B |
| Flow Rate: | 0.3 ml/min |
| Solvent A: | 50% acetonitrile/50% water; 0.1% formic acid; 5 mM ammonium formate |
| Solvent B: | 85% isopropanol/10% acetonitrile/5% water; 0.1% formic acid; 5 mM ammonium formate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002093 |
| Chromatography Summary: | Three chromatography types were employed to allow for optimal lipidome separation (i.e. C18 RPC for amphiphilic lipids, C30 RPC for triacylglycerols, HILIC for acylcarnitines). All details on the respective methods are provided in the attached methods document |
| Instrument Name: | Vanquish Horizon |
| Column Name: | Thermo Accucore C18 (150 x 2.1mm,2.6um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0-5 min - 50 to 80 % B (curve 5), 5-22 min - 80 to 95 % B (curve 4), 22-26 min - 95 % isocratic, 26-27 min - 95 to 100 % B (curve 5), 27-47 min - 100 % B isocratic, 47-47.1 min - 100 to 50 % B followed by 8 min re-equilibration at 50% B |
| Flow Rate: | 0.3 ml/min |
| Solvent A: | 50% acetonitrile/50% water; 0.1% formic acid; 5 mM ammonium formate |
| Solvent B: | 85% isopropanol/10% acetonitrile/5% water; 0.1% formic acid; 5 mM ammonium formate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002094 |
| Chromatography Summary: | Three chromatography types were employed to allow for optimal lipidome separation (i.e. C18 RPC for amphiphilic lipids, C30 RPC for triacylglycerols, HILIC for acylcarnitines). All details on the respective methods are provided in the attached methods document |
| Instrument Name: | Vanquish Horizon |
| Column Name: | Waters ACQUITY UPLC BEH HILIC (100 x 1.0mm,1.7um,130 Å) |
| Column Temperature: | 40 |
| Flow Gradient: | 0-10 min - 0 to 10 % B (curve 5), 10-10.1 min - 10 to 0 % B (curve 5) followed by 5 min re-equilibration at 0% B |
| Flow Rate: | 0.15 ml/min |
| Solvent A: | 96% acetonitrile/4% water; 5 mM ammonium acetate |
| Solvent B: | 100% water; 5 mM ammonium acetate |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002622 |
| Analysis ID: | AN002829 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap; Thermo Fusion Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | DDA and AcquireX methods for TAG identification and Parallel reaction monitoring for CE identification in nonpolar extracts in positive ion mode |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS002623 |
| Analysis ID: | AN002830 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | DDA method on polar extracts in positive and negative ion mode |
| Ion Mode: | UNSPECIFIED |
| MS ID: | MS002624 |
| Analysis ID: | AN002831 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | DDA and DDA with with inclusion list for acylcarnitines on polar extracts in positive ion mode |
| Ion Mode: | POSITIVE |
