Quantification of phosphatidic acid and lysophosphatidic acid by HPLC with evaporative light-scattering detection
- 1To whom correspondence should be addressed. e-mail: bstith{at}carbon.cudenver.edu
Abstract
Phosphatidic acid (PA) and lysophosphatidic acid (LPA) are lipids that regulate cellular processes. PA stimulates kinases and may play a role in exocytosis and membrane fusion. LPA can induce cell proliferation, platelet aggregation, and microfilament formation. Due to the growing interest in these lipids, rapid purification and quantification of these lipids is desirable. We now describe a method that utilizes one HPLC run to separate trace amounts of PA and LPA from large amounts of lipids found in cellular extracts. A two-pump HPLC with a solvent system consisting of chloroform, methanol, water, and ammonium hydroxide was employed to produce a reliable, efficient purification of the two lipids. Lipid mass was quantified by a sensitive evaporative light-scattering detector.
Using this new method, insulin addition increased both PA (87%) and LPA (217%) mass in Xenopus laevis oocytes.
- lipid
- Xenopus laevis
- oocyte
- method
- lipid extraction
- lipid signaling
- acidic phospholipids
- phospholipase D
Footnotes
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Published, JLR Papers in Press, January 16, 2003. DOI 10.1194/jlr.D200040-JLR200
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ELSD, evaporative light-scattering detection
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LPA, lysophosphatidic acid
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PA, phosphatidic acid
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PC, phosphatidylcholine
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PI, phosphatidylinositol
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PS, phosphatidylserine
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SM, sphingomyelin
Abbreviations
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- Received November 1, 2002.
- Revision received January 8, 2003.
- Copyright © 2003 by Lipid Research, Inc.
