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 JoVE Bioengineering

Cellular Lipid Uttak for Målrettet Stabil isotopfortynning Liquid Chromatography-Mass Spectrometry Analysis

1,2, 1,2, 1,2

1Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, University of Pennsylvania, 2Department of Pharmacology, University of Pennsylvania

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Cite this Article: Cellular Lipid Uttak for Målrettet Stabil isotopfortynning Liquid Chromatography-Mass Spectrometry Analysis

Gelhaus, S. L., Mesaros, A. C., Blair, I. A. Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis. J. Vis. Exp. (57), e3399, doi:10.3791/3399 (2011).

Disclosures: Cellular Lipid Uttak for Målrettet Stabil isotopfortynning Liquid Chromatography-Mass Spectrometry Analysis

Materials: Cellular Lipid Uttak for Målrettet Stabil isotopfortynning Liquid Chromatography-Mass Spectrometry Analysis

Name Company Catalog Number Comments
5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic-6,8,9,11,12,14,15-d7 acid Cayman Chemical 334250 [2H7]-5-oxoETE Internal Standard
5S-hydroxy-6E,8Z,11Z,14Z-eicosatetraenoic-5,6,8,9,11,12,14,15-d8 acid Cayman Chemical 334230 [2H8]-5(S)-HETE Internal Standard
12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic-5,6,8,9,11,12,14,15-d8 acid Cayman Chemical 334570 [2H8]-12(S)-HETE Internal Standard
15S-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic-5,6,8,9,11,12,14,15-d8 acid Cayman Chemical 334720 [2H8]-15(S)-HETE Internal Standard
20-hydroxy-5Z,8Z,11Z,14Z-eicosatetraenoic-16,16,17,17,18,18-d6 acid Cayman Chemical 390030 [2H6]-20-HETE Internal Standard
9S-hydroxy-10E,12Z-octadecadienoic-9,10,12,13-d4 acid Cayman Chemical 338410 [2H4]-9(S)-HODE Internal Standard
13S-hydroxy-9Z,11E-octadecadienoic-9,10,12,13-d4 acid Cayman Chemical 338610 [2H4]-13(S)-HODE Internal Standard
9α,11α,15S-trihydroxy-prosta-5Z,13E-dien-1-oic-17,17,18,18,19,19,20,20,20-d4 acid Cayman Chemical 316010 [2H4]-PGF2α Internal Standard
9α,11α,15S-trihydroxy-(8β)-prosta-5Z,13E-dien-1-oic-3,3,4,4-d4 acid Cayman Chemical 316350 [2H4]-8-iso-PGF2a Internal Standard
9α,11β.,15S-trihydroxy-prosta-5Z,13E-dien-1-oic-3,3,4,4-d4 acid Cayman Chemical 10008989 [2H4]-11β-PGF2 Internal Standard
9α,15S-dihydroxy-11-oxo-prosta-5Z,13E-dien-1-oic-17,17,18,18,19,19,20,20,20-d4 acid Cayman Chemical 312010 [2H4]-PGD2 Internal Standard
9-oxo-11α,15S-dihydroxy-prosta-5Z,13E-dien-1-oic-17,17,18,18,19,19,20,20,20-d4 acid Cayman Chemical 314010 [2H4]-PGE2 Internal Standard
5S,12R-dihydroxy-6Z,8E,10E,14Z-eicosatetraenoic-6,7,14,15-d4 acid Cayman Chemical 320110 [2H4]-LTB4 Internal Standard
9α,11,15S-trihydroxy-thromba-5Z,13E-dien-1-oic-3,3,4,4-d4 acid Cayman Chemical 319030 [2H4]-TxB2 Internal Standard
5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid Cayman Chemical 34250 5-oxoETE Standard
12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid Cayman Chemical 34580 12-oxoETE Standard
15-oxo-5Z,8Z,11Z,13E-eicosatetraenoic acid Cayman Chemical 34730 15-oxoETE Standard
5R-hydroxy-6E,8Z,11Z,14Z-eicosatetraenoic acid Cayman Chemical 34225 5(R)-HETE Standard
5S-hydroxy-6E,8Z,11Z,14Z-eicosatetraenoic acid Cayman Chemical 34230 5(S)-HETE Standard
8R-hydroxy-5Z,9E,11Z,14Z-eicosatetraenoic acid Cayman Chemical 34350 8(R)-HETE Standard
8S-hydroxy-5Z,9E,11Z,14Z-eicosatetraenoic acid Cayman Chemical 34360 8(S)-HETE Standard
11R-hydroxy-5Z,8Z,12E,14Z-eicosatetraenoic acid Cayman Chemical 34505 11(R)-HETE Standard
11S-hydroxy-5Z,8Z,12E,14Z-eicosatetraenoic acid Cayman Chemical 34510 11(S)-HETE Standard
12R-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid Cayman Chemical 34560 12(R)-HETE Standard
12S-hydroxy-5Z,8Z,10E,14Z-eicosatetraenoic acid Cayman Chemical 34570 12(S)-HETE Standard
15R-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic acid Cayman Chemical 34710 15(R)-HETE Standard
15S-hydroxy-5Z,8Z,11Z,13E-eicosatetraenoic acid Cayman Chemical 34720 15(S)-HETE Standard
20-hydroxy-5Z,8Z,11Z,14Z-eicosatetraenoic acid Cayman Chemical 90030 20-HETE Standard
9R-hydroxy-10E,12Z-octadecadienoic acid Cayman Chemical 38405 9(R)-HODE Standard
9S-hydroxy-10E,12Z-octadecadienoic acid Cayman Chemical 38410 9(S)-HODE Standard
13R-hydroxy-9Z,11E-octadecadienoic acid Cayman Chemical 38605 13(R)-HODE Standard
13S-hydroxy-9Z,11E-octadecadienoic acid Cayman Chemical 38610 13(S)-HODE Standard
9α,11α,15S-trihydroxy-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 16010 PGF2α Standard
9α,11α,15S-trihydroxy-(8β)-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 16350 8-iso-PGF2α Standard
9α,11β,15S-trihydroxy-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 16520 11β-PGF2 Standard
9α,15S-dihydroxy-11-oxo-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 12010 PGD2 Standard
9-oxo-11α,15S-dihydroxy-(8β)-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 14350 8-iso-PGE2 Standard
9-oxo-11α,15S-dihydroxy-prosta-5Z,13E-dien-1-oic acid Cayman Chemical 14010 PGE2 Standard
5S,12R-dihydroxy-6Z,8E,10E,14Z-eicosatetraenoic acid Cayman Chemical 20110 LTB4 Standard
9α,11,15S-trihydroxythromba-5Z,13E-dien-1-oic acid Cayman Chemical 19030 TxB2 Standard
Phosphate Buffered Saline GIBCO, by Life Technologies 14190
Diethyl Ether Sigma-Aldrich 346136
Dichloromethane Acros Organics 61030-1000 anhydrous
N,N-diisopropylethyl amine Sigma-Aldrich 387649
Pentafluorylbenzyl bromide Sigma-Aldrich 101052
Hydrochloric Acid Sigma-Aldrich 320331
Potassium Hydroxide Fluka 00650
Acetonitrile Fisher Scientific A996-4
Methanol Fisher Scientific A454-4
Chloroform Fisher Scientific 366927
Hexane Fisher Scientific H303-4
Isopropanol Fisher Scientific A464-4
Ethanol Decon Laboratories 2716
Water Fisher Scientific W7-4
Pasteur Pipets Fisher Scientific 13-678-200
10 mL Glass Centrifuge Tubes Kimble Chase 73785-10 Screw cap
Phenolic Screw Caps Kimble Chase 73802-13415
Chiralcel ADH Column Chiral Technologies 19325
HPLC vials Waters 60000751CV
HPLC inserts Waters WAT094171

References: Cellular Lipid Uttak for Målrettet Stabil isotopfortynning Liquid Chromatography-Mass Spectrometry Analysis

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