Separation and Purification of Sphingomyelin Diastereomers by High-Performance Liquid Chromatography
Identifieur interne : 003698 ( Main/Exploration ); précédent : 003697; suivant : 003699Separation and Purification of Sphingomyelin Diastereomers by High-Performance Liquid Chromatography
Auteurs : Bodil Ramstedt [Finlande] ; J. Peter Slotte [Finlande]Source :
- Analytical Biochemistry [ 0003-2697 ] ; 2000.
English descriptors
- KwdEn :
- Teeft :
- Academic press, Akademi university, Analytical hplc, Binary gradient, Biophysical properties, Chromatographic separation, Cupric acid reagent, Diastereomers, Diol column, Double band pattern, Good separation, High performance, Highperformance chromatography, Hplc, Hplc method, Hptlc, Isomer, Isomer eluting, Lipid, Lipid classes, Liquid chromatography, Mobile phase, Molecular species, Nmol, Other stereoisomer, Pergamon press, Phospholipid, Phospholipid separation, Pure isomer, Racemic, Retention times, Room temperature, Single injection, Solvent mixture, Solvent mixtures, Sphingoid base, Sphingomyelin, Sphingomyelin diastereomers, Sphingomyelin stereoisomers, Sphingomyelins, Stereoisomers, Temperature dependence.
Abstract
Abstract: All naturally occurring sphingomyelins have the d-erythro-(2S,3R) configuration of the sphingoid base. We have developed a normal-phase HPLC method for the separation of this natural stereoisomer from the l-threo-sphingomyelin, which is the other stereoisomer commonly present in semisynthetic preparations of acyl-chain defined sphingomyelins. The chromatographic method was developed by modification of a previously reported method for phospholipid separation on a normal-phase diol column. The separation was accomplished by a binary gradient of solvent mixtures (A) hexane:isopropanol:acetic acid (82:17:1.0 by vol) and (B) isopropanol:water:acetic acid (85:14:1.0 by vol) with 0.08 vol% triethylamine added to both solvent mixtures. The program of gradient elution was optimized for maximal separation of sphingomyelin diastereomers. For detection of the lipids, a light-scattering detector was used. This analytical scale HPLC method was also used for purification of the stereoisomers (up to 0.5 mg of N-oleoyl-sphingomyelin in a single injection). The purified stereoisomers were at least 99% pure according to high-performance thin-layer chromatography and analytical HPLC.
Url:
DOI: 10.1006/abio.2000.4612
Affiliations:
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Box 66, FIN 20521, Turku</wicri:regionArea><placeName><settlement type="city">Turku</settlement><region type="région" nuts="2">Finlande occidentale</region></placeName></affiliation></author></analytic><monogr></monogr><series><title level="j">Analytical Biochemistry</title><title level="j" type="abbrev">YABIO</title><idno type="ISSN">0003-2697</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2000">2000</date><biblScope unit="volume">282</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="245">245</biblScope><biblScope unit="page" to="249">249</biblScope></imprint><idno type="ISSN">0003-2697</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0003-2697</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>diastereomers</term><term>normal-phase HPLC</term><term>sphingomyelin</term><term>stereochemistry</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Academic press</term><term>Akademi university</term><term>Analytical hplc</term><term>Binary gradient</term><term>Biophysical properties</term><term>Chromatographic separation</term><term>Cupric acid reagent</term><term>Diastereomers</term><term>Diol column</term><term>Double band pattern</term><term>Good separation</term><term>High performance</term><term>Highperformance chromatography</term><term>Hplc</term><term>Hplc method</term><term>Hptlc</term><term>Isomer</term><term>Isomer eluting</term><term>Lipid</term><term>Lipid classes</term><term>Liquid chromatography</term><term>Mobile phase</term><term>Molecular species</term><term>Nmol</term><term>Other stereoisomer</term><term>Pergamon press</term><term>Phospholipid</term><term>Phospholipid separation</term><term>Pure isomer</term><term>Racemic</term><term>Retention times</term><term>Room temperature</term><term>Single injection</term><term>Solvent mixture</term><term>Solvent mixtures</term><term>Sphingoid base</term><term>Sphingomyelin</term><term>Sphingomyelin diastereomers</term><term>Sphingomyelin stereoisomers</term><term>Sphingomyelins</term><term>Stereoisomers</term><term>Temperature dependence</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: All naturally occurring sphingomyelins have the d-erythro-(2S,3R) configuration of the sphingoid base. We have developed a normal-phase HPLC method for the separation of this natural stereoisomer from the l-threo-sphingomyelin, which is the other stereoisomer commonly present in semisynthetic preparations of acyl-chain defined sphingomyelins. The chromatographic method was developed by modification of a previously reported method for phospholipid separation on a normal-phase diol column. The separation was accomplished by a binary gradient of solvent mixtures (A) hexane:isopropanol:acetic acid (82:17:1.0 by vol) and (B) isopropanol:water:acetic acid (85:14:1.0 by vol) with 0.08 vol% triethylamine added to both solvent mixtures. The program of gradient elution was optimized for maximal separation of sphingomyelin diastereomers. For detection of the lipids, a light-scattering detector was used. This analytical scale HPLC method was also used for purification of the stereoisomers (up to 0.5 mg of N-oleoyl-sphingomyelin in a single injection). The purified stereoisomers were at least 99% pure according to high-performance thin-layer chromatography and analytical HPLC.</div></front></TEI><affiliations><list><country><li>Finlande</li></country><region><li>Finlande occidentale</li></region><settlement><li>Turku</li></settlement></list><tree><country name="Finlande"><region name="Finlande occidentale"><name sortKey="Ramstedt, Bodil" sort="Ramstedt, Bodil" uniqKey="Ramstedt B" first="Bodil" last="Ramstedt">Bodil Ramstedt</name></region><name sortKey="Slotte, J Peter" sort="Slotte, J Peter" uniqKey="Slotte J" first="J. Peter" last="Slotte">J. Peter Slotte</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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