Summary of Study ST000616

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.

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Study IDST000616
Study TitleHuman Aqueous Humor Phospholipids in Control and Glaucomatous eyes
Study SummaryAn analysis of phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine and phosphatidylinositol) classes in human control and glaucomatous aqueous humor (AH). Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradford’s method, and select samples were confirmed with Densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ Quantum Access Max triple quadrupole mass spectrometer utilizing precursor ion scan (PIS) or neutral ion loss scan (NLS) using appropriate class specific lipid standards in a two-step quantification process. Mass spectrometer data were analyzed using MzMine 2.23.
Institute
University of Miami
Last NameBhattacharya
First NameSanjoy
Address1638 NW 10th Avenue, Miami, Florida, 33136, USA
Emailsbhattacharya@med.miami.edu
Phone3054824103
Submit Date2017-05-31
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailMS(Dir. Inf.)
Release Date2019-07-17
Release Version1
Sanjoy Bhattacharya Sanjoy Bhattacharya
https://dx.doi.org/10.21228/M8WC8S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000450
Project DOI:doi: 10.21228/M8WC8S
Project Title:Phospholipid Profiling of Human Glaucomatous and Control Aqueous Humor and Trabecular Meshwork
Project Summary:In this study we used a triple quadrupole mass spectrometer to investigate the phospholipids within human aqueous humor and trabecular meshwork. Our aim was to find potential differences in the lipid profiles between control and glaucomatous donors to better understand the disease.
Institute:University of Miami
Last Name:Bhattacharya
First Name:Sanjoy
Address:1638 NW 10th Avenue, Miami, Florida, 33136, USA
Email:sbhattacharya@med.miami.edu
Phone:3054824103

Subject:

Subject ID:SU000639
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Condition
SA034156NAH16L1Control
SA034157NAH14L1Control
SA034158NAH13L1Control
SA034159NAH17L1Control
SA034160NAH19L1Control
SA034161NAH1BL1Control
SA034162NAH20L1Control
SA034163NAH12L1Control
SA034164NAH18L1Control
SA034165NAH15L1Control
SA034166NAH5BL1Control
SA034167NAH4BL1Control
SA034168NAH11L1Control
SA034169NAH2BL1Control
SA034170NAH6BL1Control
SA034171NAH3BL1Control
SA034172NAH7BL1Control
SA034173NAH10BL1Control
SA034174NAH9BL1Control
SA034175NAH8BL1Control
SA034176GAH15L1Glaucomatous
SA034177GAH14L1Glaucomatous
SA034178GAH13L1Glaucomatous
SA034179GAH16L1Glaucomatous
SA034180GAH19L1Glaucomatous
SA034181GAH12L1Glaucomatous
SA034182GAH20L1Glaucomatous
SA034183GAH18L1Glaucomatous
SA034184GAH17L1Glaucomatous
SA034185GAH1BL1Glaucomatous
SA034186GAH5BL1Glaucomatous
SA034187GAH4BL1Glaucomatous
SA034188GAH3BL1Glaucomatous
SA034189GAH2BL1Glaucomatous
SA034190GAH6BL1Glaucomatous
SA034191GAH7BL1Glaucomatous
SA034192GAH10BL1Glaucomatous
SA034193GAH9BL1Glaucomatous
SA034194GAH8BL1Glaucomatous
SA034195GAH11L1Glaucomatous
Showing results 1 to 40 of 40

Collection:

Collection ID:CO000633
Collection Summary:Control and POAG AH were procured during glaucoma, cataract surgeries following institutional review board approved protocols, and principles outlined in the Declaration of Helsinki were adhered to. A total of 20 control and 20 glaucomatous AH samples were included for these studies. All AH samples were immediately stored at −80°C until time of use. The mean age of donors was 69.5 ± 9.9 years and both genders were included for these studies.
Sample Type:Eye tissue

Treatment:

Treatment ID:TR000653
Treatment Summary:No treatment was performed on the eyes.

Sample Preparation:

Sampleprep ID:SP000646
Sampleprep Summary:Aqueous humor samples were subjected to extraction of lipids using suitable and minimal modification of Bligh and Dyer method. The lower organic phase containing the extracted lipids was isolated and solvent dried with a Speed-Vac (Model 7810014; Labconco, Kansas City, MO). Samples were subsequently flushed with argon gas to prevent oxidation. Proteins recovered from the corresponding upper aqueous phase were quantified using the Bradford method. A subset of protein samples were also subjected to densitometric quantification using bovine serum albumin (BSA) as a standard (amino acid quantified) after electrophoretic separation on a PHAST (GE Healthcare Bio-Sciences AB, Sweden) gel system. We repeated protein estimations using an amino acid analyzer after overnight digestion in hydrochloric acid following previously published protocols. The protein amounts determined using amino acid analyzer were utilized in normalization of lipids per amount of proteins. In order to determine and ensure extraction efficiency, ovine wool cholesterol (molecular mass 386.7; catalog no. 700000; Avanti Polar Lipids, Albaster, AL) was premixed with AH prior to extraction. All extractions and subsequent handling were made using glass vials to avoid contaminating impurities. The quantification lipid standards (all procured from Avanti Polar Lipids, Albaster, AL) we used were ditridecanoyl-sn-glycero-3-phosphocholine (molecular mass 649.89, catalog no. 850340) for PCs, 1,2-dioleoyl-snglycero- 3-phospho-L-serine (molecular mass 810.03, catalog no. 840035) for PSs, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (molecular mass 744.04, catalog no. 850725) for PEs and 1,2-dioleoyl-sn-glycero-3-phospho-(10-myo-inositol) (molecular mass 880.15, catalog no. 850149) for PIs.

Combined analysis:

Analysis ID AN000945
Analysis type MS
Chromatography type None (Direct infusion)
Chromatography system None
Column none
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Thermo Quantiva Access Max
Ion Mode UNSPECIFIED
Units Peak Area

Chromatography:

Chromatography ID:CH000673
Instrument Name:None
Column Name:none
Chromatography Type:None (Direct infusion)

MS:

MS ID:MS000841
Analysis ID:AN000945
Instrument Name:Thermo Quantiva Access Max
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:UNSPECIFIED
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