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JoVE Journal Bioengineering
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Bioengineering

Cellular Lipid Ekstraktion for målrettet stabile isotoper Fortynding Liquid Chromatography-massespektrometri Analyse

Published: November 17, 2011 doi: 10.3791/3399
Automatic Translation
1,2, 1,2, 1,2
1Centers for Cancer Pharmacology and Excellence in Environmental Toxicology, University of Pennsylvania, 2Department of Pharmacology, University of Pennsylvania

Materials

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-ox–TE 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-ox–TE Standard
12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid Cayman Chemical 34580 12-ox–TE Standard
15-oxo-5Z,8Z,11Z,13E-eicosatetraenoic acid Cayman Chemical 34730 15-ox–TE 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

DOWNLOAD MATERIALS LIST

References

  1. Needleman, P., Turk, J., Jakschik, B. A., Morrison, A. R., Lefkowith, J. B. Arachidonic acid metabolism. Annu. Rev. Biochem. 55, 69-102 (1986).
  2. Nikolaev, V., Reddanna, P., Whelan, J., Hildenbrandt, G., Reddy, C. C. Stereochemical nature of the products of linoleic acid oxidation catalyzed by lipoxygenases from potato and soybean. Biochem. Biophys. Res. Commun. 170, 491-496 (1990).
  3. Panigrahy, D., Kaipainen, A., Greene, E. R., Huang, S. Cytochrome P450-derived eicosanoids: the neglected pathway in cancer. Cancer. Metastasis. Rev. 29, 723-735 (2010).
  4. Basu, S. Bioactive eicosanoids: role of prostaglandin F(2alpha) and F-isoprostanes in inflammation and oxidative stress related pathology. Mol. Cells. 30, 383-391 (2010).
  5. Sanchez-Mejia, R. O., Mucke, L. Phospholipase A2 and arachidonic acid in Alzheimer's disease. Biochim. Biophys. Acta. 1801, 784-790 (2010).
  6. Hersberger, M. Potential role of the lipoxygenase derived lipid mediators in atherosclerosis: leukotrienes, lipoxins and resolvins. Clin. Chem. Lab Med. 48, 1063-1073 (2010).
  7. Allayee, H., Roth, N., Hodis, H. N. Polyunsaturated fatty acids and cardiovascular disease: implications for nutrigenetics. J. Nutrigenet. Nutrigenomics. 2, 140-148 (2009).
  8. Capdevila, J. H., Falck, J. R. Biochemical and molecular properties of the cytochrome P450 arachidonic acid monooxygenases. Prostaglandins Other Lipid Mediat. , 68-69 (2002).
  9. Ciccimaro, E., Blair, I. A. Stable-isotope dilution LC-MS for quantitative biomarker analysis. Bioanalysis. 2, 311-341 (2010).
  10. Blair, I. A. Electron-capture negative-ion chemical ionization mass-spectrometry of lipid mediators. Methods in Enzymology. 187, 13-23 (1990).
  11. Singh, G., Gutierrez, A., Xu, K., Blair, I. A. Liquid chromatography/electron capture atmospheric pressure chemical ionization/mass spectrometry: analysis of pentafluorobenzyl derivatives of biomolecules and drugs in the attomole range. Analytical Chemistry. 72, 3007-3013 (2000).
  12. Blair, I. A. Encyclopedia of Mass Spectrometry. Capriol, R. M., Gross, M. L. , Elsevier. 283-307 (2005).
  13. Lee, S. H., Blair, I. A. Targeted chiral lipidomics analysis by liquid chromatography electron capture atmospheric pressure chemical ionization mass spectrometry (LC-ECAPCI/MS). Methods in Enzymology. 433, 159-174 (2007).
Cellular Lipid Ekstraktion for målrettet stabile isotoper Fortynding Liquid Chromatography-massespektrometri Analyse
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Cite this Article

Gelhaus, S. L., Mesaros, A. C.,… More

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).

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