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MB Sample ID: SA326759

Local Sample ID:	N13
Subject ID:	SU003116
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Male and female

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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.

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.