A sensitive, radiometric assay for lysophosphatidylcholine

David J. Dobmeyer; Peter B. Corr; Michael H. Creer

doi:10.1016/0003-2697(90)90251-4

A sensitive, radiometric assay for lysophosphatidylcholine

David J. Dobmeyer, Peter B. Corr, Michael H. Creer

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Abstract

To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [³H]acetic anhydride and the [³H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [¹⁴C]lysophosphatidylcholine as an internal standard.

Original language	English (US)
Pages (from-to)	36-43
Number of pages	8
Journal	Analytical Biochemistry
Volume	185
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(90)90251-4
State	Published - Feb 15 1990

All Science Journal Classification (ASJC) codes

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0003-2697(90)90251-4

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@article{5af758fcca144a5ab348a7ab30f01047,

title = "A sensitive, radiometric assay for lysophosphatidylcholine",

abstract = "To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [3H]acetic anhydride and the [3H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [14C]lysophosphatidylcholine as an internal standard.",

author = "Dobmeyer, {David J.} and Corr, {Peter B.} and Creer, {Michael H.}",

note = "Funding Information: 1 Research from the authors{\textquoteright} laboratory was supported National Institutes of Health Grant HL 17646, SCOR Heart Disease and NIH Grants HL 28995 and HL 36773. 2 Abbreviations used: LPC, lysophosphatidylcholine; acid methyl ester; DMAP, dimethylaminopyridine; [%]palmitoyl)-2-lysophosphatidylcholine.",

year = "1990",

month = feb,

day = "15",

doi = "10.1016/0003-2697(90)90251-4",

language = "English (US)",

volume = "185",

pages = "36--43",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

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T1 - A sensitive, radiometric assay for lysophosphatidylcholine

AU - Dobmeyer, David J.

AU - Corr, Peter B.

AU - Creer, Michael H.

N1 - Funding Information: 1 Research from the authors’ laboratory was supported National Institutes of Health Grant HL 17646, SCOR Heart Disease and NIH Grants HL 28995 and HL 36773. 2 Abbreviations used: LPC, lysophosphatidylcholine; acid methyl ester; DMAP, dimethylaminopyridine; [%]palmitoyl)-2-lysophosphatidylcholine.

PY - 1990/2/15

Y1 - 1990/2/15

N2 - To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [3H]acetic anhydride and the [3H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [14C]lysophosphatidylcholine as an internal standard.

AB - To facilitate investigation of the metabolism of lysophosphatidylcholine and choline lysoplasmalogen in small quantities of tissue, a method for the quantification of these phospholipid species that is capable of accurate and reproducible analysis in samples which contain less than 1 nmol of total choline lysophospholipid was developed. The procedure employs chloroform and methanol extraction of phospholipids from isolated tissue with subsequent separation of the choline lysophospholipid fraction by high-performance liquid chromatography. The choline lysophospholipids are then acetylated with [3H]acetic anhydride and the [3H]acetyl-lysophosphatidylcholine product is isolated by thin-layer chromatography and quantified by liquid scintillation counting. The choline lysophospholipid content in the sample is determined from a standard curve constructed from samples containing a known amount of synthetic lysophosphatidylcholine with correction for recovery based on the inclusion of [14C]lysophosphatidylcholine as an internal standard.

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

IS - 1

ER -

A sensitive, radiometric assay for lysophosphatidylcholine

Abstract

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