Quantification of phosphatidic acid and lysophosphatidic acid by HPLC with evaporative light-scattering detection

William L. Holland, Erinn C. Stauter, Bradley J. Stith

Research output: Contribution to journalArticle

24 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)854-858
Number of pages5
JournalJournal of Lipid Research
Volume44
Issue number4
DOIs
StatePublished - Apr 2003

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Phosphatidic Acids
Light scattering
High Pressure Liquid Chromatography
Lipids
Light
Purification
Ammonium Hydroxide
Membrane Fusion
Exocytosis
Xenopus laevis
Cell proliferation
Chloroform
Platelets
Actin Cytoskeleton
Platelet Aggregation
Oocytes
Methanol
lysophosphatidic acid
Fusion reactions
Agglomeration

Keywords

  • Acidic phospholipids
  • Lipid
  • Lipid extraction
  • Lipid signaling
  • Method
  • Oocyte
  • Phospholipase D
  • Xenopus laevis

ASJC Scopus subject areas

  • Endocrinology

Cite this

Quantification of phosphatidic acid and lysophosphatidic acid by HPLC with evaporative light-scattering detection. / Holland, William L.; Stauter, Erinn C.; Stith, Bradley J.

In: Journal of Lipid Research, Vol. 44, No. 4, 04.2003, p. 854-858.

Research output: Contribution to journalArticle

Holland, William L. ; Stauter, Erinn C. ; Stith, Bradley J. / Quantification of phosphatidic acid and lysophosphatidic acid by HPLC with evaporative light-scattering detection. In: Journal of Lipid Research. 2003 ; Vol. 44, No. 4. pp. 854-858.
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