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MB Sample ID: SA326789
| Local Sample ID: | PX15 |
| Subject ID: | SU003116 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003116 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| PX15 | SA326789 | FL038821 | PEXG | Factor |
Collection:
| Collection ID: | CO003109 |
| Collection Summary: | All AH samples collected came from human donors during cataract surgery. A total of 66 samples were collected which included non-glaucomatous controls and those with pseudoexfoliation syndrome (PEX), pseudoexfoliation glaucoma (PEXG), and primary open-angle glaucoma (POAG). The clinicians provided samples from the Department of Ophthalmology of Hospital Clínico San Carlos in Madrid, Spain. The patient samples consisted of 19 POAG, 9 PEX, 14 PEXG, and 23 controls. |
| Sample Type: | Eye tissue |
Treatment:
| Treatment ID: | TR003125 |
| Treatment Summary: | No treatment |
Sample Preparation:
| Sampleprep ID: | SP003122 |
| Sampleprep Summary: | Lipids were extracted from POAG, PEX, PEXG, and control AH samples using the Bligh and Dyer method (Bligh and Dyer, 1959). EquiSPLASHTM Lipidomix quantitative mass spec internal standard was spiked into each sample to normalize the lipids. For the extraction, a 2:1 chloroform:methanol solution with 100 µL of water was added to promote phase separation followed by centrifugation at four °C at 14,000 rpm for 20 minutes. After centrifugation, separation was observed and the extracted lipids were collected from the lower, organic chloroform phase with a syringe. Once the lipids were isolated and transferred to 2 mL glass vials, they were dried in a Speed-Vac for about 90 minutes. The upper, aqueous, water/methanol phase containing most of the proteins was collected and stored for a Bradford protein assay to measure protein concentration in the future. When the samples were dehydrated, they were flushed with argon gas for storage at -80°C until ready for mass spec analysis. To prepare the samples for mass spec analysis, the dried lipids were resuspended in a 1:1 acetonitrile:isopropyl alcohol solution, sonicated for 30 minutes, and aliquoted into mass spec vials. |
Combined analysis:
| Analysis ID | AN004932 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Vanquish Horizon Binary UHPLC |
| Column | Accucore Vanquish C18+ UHPLC (2.1mm x 150mm x 1.5µm) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | µg/mL |
Chromatography:
| Chromatography ID: | CH003722 |
| Instrument Name: | Vanquish Horizon Binary UHPLC |
| Column Name: | Accucore Vanquish C18+ UHPLC (2.1mm x 150mm x 1.5µm) |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS004675 |
| Analysis ID: | AN004932 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Lipids were identified and quantified with LipidSearch 4.2.21. Statistical analysis was conducted through MetaboAnalyst 5.0. |
| Ion Mode: | UNSPECIFIED |
