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MB Sample ID: SA214237
Local Sample ID: | Control_Negative_P-06 |
Subject ID: | SU002329 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU002329 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Control_Negative_P-06 | SA214237 | FL026025 | Control | Group |
Collection:
Collection ID: | CO002322 |
Collection Summary: | Venous blood was drawn in 8.5 mL purple cap Vacutainer EDTA tubes and gently invert to mix. Plasma was collected after centrifugation at 1000 g for 15 min. All samples were immediately aliquoted to Eppendorf tubes and frozen at -80 ºC until analysis. |
Sample Type: | Blood (plasma) |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR002341 |
Treatment Summary: | NA |
Sample Preparation:
Sampleprep ID: | SP002335 |
Sampleprep Summary: | 50 μL plasma, was mixed with 20 μL internal standard working solution (SPLASH® LIPIDOMIX® Mass Spec Standard + Cer/sph mixture I), extracted with 1000 μL butanol/methanol (1/1, v/v, 10 mM ammonium formate) for 10 min on a vortex mixture. Samples were pelleted by centrifugation at 4000 x g for 5 min at room temperature. The supernatant was moved to a clean glass tube, dried under nitrogen, and resuspended in 100 μL MTBE/methanol (1/3, v/v) with 10 mM ammonium formate. 5 μL of each sample was combined to make a pooled quality control (QC) sample and ran every ten samples in the long sequence to monitor retention time and signal intensity drift. The remaining samples (50 μL) were transferred into injection vials for lipidomic analysis. |
Combined analysis:
Analysis ID | AN003661 | AN003662 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
Column | Thermo Accucore C18 (100 x 2.1mm,2.6um) | Thermo Accucore C18 (100 x 2.1mm,2.6um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | intensity | intensity |
Chromatography:
Chromatography ID: | CH002713 |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Thermo Accucore C18 (100 x 2.1mm,2.6um) |
Column Temperature: | 35 |
Flow Gradient: | 0 min, 90% A; 1 min, 90% A; 4 min, 60% A; 12 min, 25% A; 21 min, 1% A; 24 min, 1% A; 24.1 min, 90% A; 28 min, 90%. |
Flow Rate: | 0.4 ml/min |
Injection Temperature: | 4 |
Solvent A: | 50% acetonitrile/50% water; 0.1% formic acid; 10 mM ammonium formate |
Solvent B: | 10% acetonitrile/88% isopropanol/2% water; 0.02% formic acid; 2 mM ammonium formate |
Analytical Time: | 28min |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003412 |
Analysis ID: | AN003661 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Samples were analyzed using a Q Exactive HF (QE-HF) (Thermo Scientific, Waltham, MA) equipped with a heated electro-spray ionization (HESI) source operated in both positive and negative ion mode. For DDA library generation (untargeted lipidomics), data acquisition was performed on the pooled QC samples in Full Scan/ddMS2 mode @ 120,000 resolutions. The Full Scan settings as follows: AGC target = 1e6; Maximum IT = 250 ms; scan range = 250 to 1800 m/z. Top 20 MS/MS spectral (dd-MS2) @ 15000 were generated with AGC target = 1e5, Maximum IT=25 ms, and (N)CE/stepped nce = 20, 30, 40v. For untargeted lipid analysis, LipidSearch 4.2 (Thermo Scientific, Waltham, MA) was used for peak detection, identification, alignment, and quantification. Well annotated lipids (MS spectral match) were used to generate a inclusion list (Supplementary Table 4) for targeted lipidomics analysis. Targeted lipidomics were performed in Full scan + PRM modes. The PRM settings were as follows: resolution = 15000, AGC target = 2e5, Maximum IT=25 ms, loop count = 36, isolation window = 2.0, (N)CE was optimized for each lipid class. |
Ion Mode: | POSITIVE |
MS ID: | MS003413 |
Analysis ID: | AN003662 |
Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Samples were analyzed using a Q Exactive HF (QE-HF) (Thermo Scientific, Waltham, MA) equipped with a heated electro-spray ionization (HESI) source operated in both positive and negative ion mode. For DDA library generation (untargeted lipidomics), data acquisition was performed on the pooled QC samples in Full Scan/ddMS2 mode @ 120,000 resolutions. The Full Scan settings as follows: AGC target = 1e6; Maximum IT = 250 ms; scan range = 250 to 1800 m/z. Top 20 MS/MS spectral (dd-MS2) @ 15000 were generated with AGC target = 1e5, Maximum IT=25 ms, and (N)CE/stepped nce = 20, 30, 40v. For untargeted lipid analysis, LipidSearch 4.2 (Thermo Scientific, Waltham, MA) was used for peak detection, identification, alignment, and quantification. Well annotated lipids (MS spectral match) were used to generate a inclusion list (Supplementary Table 4) for targeted lipidomics analysis. Targeted lipidomics were performed in Full scan + PRM modes. The PRM settings were as follows: resolution = 15000, AGC target = 2e5, Maximum IT=25 ms, loop count = 36, isolation window = 2.0, (N)CE was optimized for each lipid class. |
Ion Mode: | NEGATIVE |