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MB Sample ID: SA322377
| Local Sample ID: | 20210625_AJU_Y3_lipidomics |
| Subject ID: | SU003073 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003073 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 20210625_AJU_Y3_lipidomics | SA322377 | FL038467 | Test sample | Sample_type |
Collection:
| Collection ID: | CO003066 |
| Collection Summary: | The experiment was initiated by seeding the cells in a 100mm cell culture dish, followed by a 2-day incubation period, during which the culture medium was carefully harvested from the cell culture dish. Use a sterile technique to minimize contamination. Transfer the harvested culture medium to a suitable container. |
| Collection Protocol Filename: | Sample_collection_lipid.pdf |
| Sample Type: | Cultured fibroblasts |
Treatment:
| Treatment ID: | TR003082 |
| Treatment Summary: | no treatment |
Sample Preparation:
| Sampleprep ID: | SP003079 |
| Sampleprep Summary: | For lipid extraction from samples, a two-step method involving neutral and acidic extraction were used. At first, in neutral extraction, lipids from the samples were extracted according to the Folch method using a mixture of chloroform and methanol (2:1, v/v). The samples were vortexed and incubated on ice for 10 min. The samples were centrifuged at 13,800×g for 2 min at 4°C, supernatant was transferred to a new 1.5 mL tube. Next, in Acidic extraction, 750 μL of chloroform:methanol:HCl (1N, 37%) (40:80:1, v/v/v) was added to the remaining samples. After incubating for 15 min at room temperature, 250 μL of cold chloroform and 450 μL of cold 0.1 M HCl were added to the sample. The mixture was vortexed for 1 min and centrifuged at 6,500×g for 2 min at 4°C. The lower organic phase was collected and combined with the prior extract. The sample was then dried using HyperVAC-MAX VC2200 centrifugal vacuum concentrator (Hanil Scientific Inc., Korea). Dried metabolite contents were reconstituted in 50 µL of mobile phase solvent A:solvent B (2 :1, v/v) and then subjected to LC-MS/MS analysis. |
| Sampleprep Protocol Filename: | Sample_prep_lipid.pdf |
Combined analysis:
| Analysis ID | AN004861 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity UHPLC |
| Column | Thermo Hypersil GOLD C18 (100 x 2.1mm,1.9um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Q Exactive™ Hybrid Quadrupole-Orbitrap MS |
| Ion Mode | POSITIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH003669 |
| Methods Filename: | LC_MS_Method_lipid.pdf |
| Instrument Name: | Agilent 1290 Infinity UHPLC |
| Column Name: | Thermo Hypersil GOLD C18 (100 x 2.1mm,1.9um) |
| Column Temperature: | 320℃ |
| Flow Gradient: | 0–5 min, 5% B; 5–15 min, 5–30% B; 15–22 min, 30–90% B; 22–25 min, 90% B; 25–26 min, 90–5% B; 26–30 min, 5% B. Parameters were set as follows: positive mode, spray voltage; 3.8 kV |
| Flow Rate: | 0.2 mL/min |
| Solvent A: | Acetonitrile:Methanol:Water (19:19:2, v/v/v) + 20 mmol/L ammonium formate + 0.1% formic acid (v/v) |
| Solvent B: | 2-propanol + 20 mmol/L ammonium formate + 0.1% formic acid (v/v) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004605 |
| Analysis ID: | AN004861 |
| Instrument Name: | Q Exactive™ Hybrid Quadrupole-Orbitrap MS |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Obtained UHPLC-Orbitrap-MS/MS RAW files were processed using Lipid Search 4.2TM (Thermo Fisher Scientific, Waltham, MA, USA). Lipid profiling under the following conditions: parent search; 0.2 Da, product search; 5.0 ppm, precursor ion mass tolerance; 8.0 ppm, M-score threshold; 2.0. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | MS_Analysis_lipid.pdf |
