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MB Sample ID: SA169297
| Local Sample ID: | ODBS1_1 |
| Subject ID: | SU001901 |
| 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: | SU001901 |
| 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 |
|---|---|---|---|---|
| ODBS1_1 | SA169297 | FL020195 | ODBS | Genotype |
Collection:
| Collection ID: | CO001894 |
| 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: | TR001914 |
| Treatment Summary: | 10 µl of each serum sample were added to 10 µl of 0.9% NaCl and 120 µl of CHCl3: MeOH (2:1, v/v) containing a 2.5 ppm solution of different lipids standards. Lipids were extracted with CHCl3: MeOH (2:1, v/v). |
Sample Preparation:
| Sampleprep ID: | SP001907 |
| Sampleprep Summary: | 10 µl of each serum sample were added to 10 µl of 0.9% NaCl and 120 µl of CHCl3: MeOH (2:1, v/v) containing a 2.5 ppm solution of different lipids standards. Samples were randomized, vortex-mixed and put on ice for 30 min. After that, samples were centrifuged (9400 g, 3 min, 4 °C). Then, 60 µl of the lower layer of each sample was transferred to a glass vial with an insert and 60 µl of CHCl3: MeOH (2:1, v/v) was added. Samples were stored at -80 °C until LC-MS analysis. |
| Processing Storage Conditions: | On ice |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002960 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent Infinity 1290 |
| Column | Waters ACQUITY UPLC BEH C18 |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6545 Q-TOF |
| Ion Mode | POSITIVE |
| Units | Area |
Chromatography:
| Chromatography ID: | CH002193 |
| Chromatography Summary: | An ultra-high-performance liquid chromatography electrospray ionization quadrupole time-of-flight (UHPLC-ESI-Q-TOF-MS) were used to analyze the samples on positive ionization mode. Samples were analyzed by duplicate. UHPLC system was an Agilent Infinity 1290 provided by Agilent Technologies (Santa Clara, CA, USA) equipped with a multisampler (kept at -10 °C). Needle wash solutions were performed with 10% DCM in MeOH and ACN:MeOH:IPA:H2O (1:1:1:1, v/v/v/v), and 0.1% of HCOOH after each injection for 7.5 s. System were equipped with a column thermostat (maintained at 50 °C) and a quaternary solvent manager. An ACQUITY UPLC BEH C18 column was used for separations (2.1 mm x 100 mm, particle size 1.7 μm) purchased at Waters (Milford, CT, USA). Injection volume was 1 μl and flow rate was established at 0.4 ml/min. Mobile phases were composed of (A) H2O + NH4AC 10 mM + 0.1 % HCOOH and (B) ACN: IPA (1:1, v/v) + NH4AC 10 mM + 0.1 % HCOOH. Gradient was from 0 to 2 min 35-80 % B, 2 min to 7 min 80-100 % B and 7 to 14 min 100 % B. A re-equilibration of 7 min was performed after each run to bring system to initial conditions (35 % B). |
| Instrument Name: | Agilent Infinity 1290 |
| Column Name: | Waters ACQUITY UPLC BEH C18 |
| Column Temperature: | 50 |
| Flow Gradient: | 0 to 2 min 35-80 % B, 2 min to 7 min 80-100 % B and 7 to 14 min 100 % B |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 100% water; 0.1% formic acid; 10 mM ammonium acetate |
| Solvent B: | 50% acetonitrile/50% isopropanol; 0.1% formic acid; 10 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002750 |
| Analysis ID: | AN002960 |
| Instrument Name: | Agilent 6545 Q-TOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Mass Spectrometer was an Agilent 6545 quadrupole time-of-flight (Q-TOF) mounted with a dual jet stream electrospray (dual ESI) ion source interface. Nitrogen was obtained from a nitrogen generator (PEAK Scientific, Renfrewshire, Scotland, UK) as a sheath gas at a flow rate of 11 l/min at 379 °C. As a Collision gas was used Pure Nitrogen (6.0) from Praxair (Fredericia, Denmark). Capillary voltage was maintained at 3600 V and nozzle voltage was kept at 1500 V. Reference mass solution was prepared in consonance with Agilent guidelines, including ions at m/z 121.0509 and 922.0098. The second nebulizer was used to introduce the solution in the dual ESI ion source through the isocratic pump at a constant flow rate of 4 ml/min (split to 1:100 before nebulization). Acquisition mass range was 100 to 3000 m/z. Instrument used the extended dynamic range with an estimated resolution of 30.000 FWHM measured at 1521.9715 m/z (included in tune mixture) when instrument calibration was performed. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3600 V |
