Summary of Study ST000624

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000450. The data can be accessed directly via it's Project DOI: 10.21228/M8WC8S This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

Study ID	ST000624
Study Title	Human trabecular meshowrk phospholipid profiles of control and glaucomatous eyes
Study Summary	We compared phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol) profiles of control and glaucomatous trabecular meshwork (TM) derived from human donors.Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradford method, and for select samples confirmed with densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ quantum Access Max triple quadrupole mass spectrometer with precursor ion scan (PIS) or neutral ion loss scan (NLS), using appropriate class specific lipid standards.
Institute	University of Miami
Last Name	Bhattacharya
First Name	Sanjoy
Address	McKnight Vision Research Building, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, 1638 NW 10th Avenue, Room 706A, Miami, Florida 33136
Email	SBhattacharya@med.miami.edu
Phone	305-482-4103
Submit Date	2017-06-01
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	MS(Dir. Inf.)
Release Date	2019-07-17
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP000654
Sampleprep Summary:	Isolated trabecular meshwork tissue was subjected to an alternating cycle of immersion in liquid nitrogen and 37˚C, followed by extraction of lipids using a modified Bligh and Dryer method.13 The organic phase with extracted lipids was dried in a Speed-Vac (Model 7810014; Labconco, Kansas City, MO). Samples were flushed with argon gas to prevent oxidation. Corresponding aqueous phase extracted proteins were subjected to determination of concentration using Bradford’s method. A subset of protein samples also were subjected to densitometric quantification using bovine serum albumin as a standard (amino acid quantified) after electrophoretic separation on a PHAST (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) gel system. All extractions and subsequent handling were made using glass vials, and polyvinyl plastic was avoided completely. We also added a PC control standard during tissue homogenization, determined its recovery in an aliquot, and used for the calculation of total recovery of this standard for each extraction to ensure >99% recovery of added standard during extraction.

Sampleprep ID:

SP000654

Sampleprep Summary:

Isolated trabecular meshwork tissue was subjected to an alternating cycle of immersion in liquid nitrogen and 37˚C, followed by extraction of lipids using a modified Bligh and Dryer method.13 The organic phase with extracted lipids was dried in a Speed-Vac (Model 7810014; Labconco, Kansas City, MO). Samples were flushed with argon gas to prevent oxidation. Corresponding aqueous phase extracted proteins were subjected to determination of concentration using Bradford’s method. A subset of protein samples also were subjected to densitometric quantification using bovine serum albumin as a standard (amino acid quantified) after electrophoretic separation on a PHAST (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) gel system. All extractions and subsequent handling were made using glass vials, and polyvinyl plastic was avoided completely. We also added a PC control standard during tissue homogenization, determined its recovery in an aliquot, and used for the calculation of total recovery of this standard for each extraction to ensure >99% recovery of added standard during extraction.