Major human plasma lipid classes determined by quantitative high-performance liquid chromatography, their variation and associations with phospholipid fatty acids

Tuulikki Seppänen-Laakso (Corresponding Author), I. Laakso, H. Vanhanen, K. Kiviranta, T. Lehtimäki, R. Hiltunen

Research output: Contribution to journalArticleScientificpeer-review

43 Citations (Scopus)

Abstract

An HPLC method with evaporative light-scattering detection (ELSD) was optimized and validated for the simultaneous quantitation of cholesteryl esters (CEs), triacylglycerols (TGs), free cholesterol (FC) and phosphatidylcholine (PC) in human plasma. The separation of CEs from TGs, the most variable plasma lipid class, was improved by speeding up the gradient steps and by increasing the re-equilibration time between runs. The calibrations were made at levels of 0.14–14 μg lipid/injection. The intra- and inter-day precision values of the method ranged between 1.9 and 4.5 and 2.3–7.2% (RSD, n=6), respectively, including determinations at two concentration levels. In comparison to other lipid classes, quantitation of PC proved to be equally repeatable despite its lowest detector response. The relative recoveries varied from 97.0 to 110.3%, showing good accuracy of the method. The methodological variation of the lipid classes covered 0.6–3.1% of their total variation in the study population (n=48). The CE/FC ratio showed an even closer relationship with phospholipid linoleic acid (18:2n−6; r=0.65, P<0.001) than with serum cholesterol levels, while eicosapentaenoic acid (20:5n−3) was significantly associated with PC (r=0.41, P<0.01). The CE/FC ratio increased (P<0.01) during soyabean oil substitution and the level of PC increased (P<0.01) during cold-pressed rapeseed oil substitution.
Original languageEnglish
Pages (from-to)437-445
Number of pages9
JournalJournal of Chromatography B: Biomedical Sciences and Applications
Volume754
Issue number2
DOIs
Publication statusPublished - 2001
MoE publication typeA1 Journal article-refereed

Fingerprint

Plasma (human)
Cholesterol Esters
High performance liquid chromatography
Phosphatidylcholines
Phospholipids
Fatty Acids
Cholesterol
High Pressure Liquid Chromatography
Lipids
Triglycerides
Substitution reactions
Eicosapentaenoic Acid
Linoleic Acid
Light scattering
Calibration
Oils
Detectors
Plasmas
Light
Recovery

Keywords

  • Cholesteryl esters
  • Triacylglycerols
  • Free cholesterol
  • Phosphatidylcholine
  • Phospholipid fatty acids

Cite this

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title = "Major human plasma lipid classes determined by quantitative high-performance liquid chromatography, their variation and associations with phospholipid fatty acids",
abstract = "An HPLC method with evaporative light-scattering detection (ELSD) was optimized and validated for the simultaneous quantitation of cholesteryl esters (CEs), triacylglycerols (TGs), free cholesterol (FC) and phosphatidylcholine (PC) in human plasma. The separation of CEs from TGs, the most variable plasma lipid class, was improved by speeding up the gradient steps and by increasing the re-equilibration time between runs. The calibrations were made at levels of 0.14–14 μg lipid/injection. The intra- and inter-day precision values of the method ranged between 1.9 and 4.5 and 2.3–7.2{\%} (RSD, n=6), respectively, including determinations at two concentration levels. In comparison to other lipid classes, quantitation of PC proved to be equally repeatable despite its lowest detector response. The relative recoveries varied from 97.0 to 110.3{\%}, showing good accuracy of the method. The methodological variation of the lipid classes covered 0.6–3.1{\%} of their total variation in the study population (n=48). The CE/FC ratio showed an even closer relationship with phospholipid linoleic acid (18:2n−6; r=0.65, P<0.001) than with serum cholesterol levels, while eicosapentaenoic acid (20:5n−3) was significantly associated with PC (r=0.41, P<0.01). The CE/FC ratio increased (P<0.01) during soyabean oil substitution and the level of PC increased (P<0.01) during cold-pressed rapeseed oil substitution.",
keywords = "Cholesteryl esters, Triacylglycerols, Free cholesterol, Phosphatidylcholine, Phospholipid fatty acids",
author = "Tuulikki Sepp{\"a}nen-Laakso and I. Laakso and H. Vanhanen and K. Kiviranta and T. Lehtim{\"a}ki and R. Hiltunen",
year = "2001",
doi = "10.1016/S0378-4347(01)00031-7",
language = "English",
volume = "754",
pages = "437--445",
journal = "Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences",
issn = "1570-0232",
publisher = "Elsevier",
number = "2",

}

Major human plasma lipid classes determined by quantitative high-performance liquid chromatography, their variation and associations with phospholipid fatty acids. / Seppänen-Laakso, Tuulikki (Corresponding Author); Laakso, I.; Vanhanen, H.; Kiviranta, K.; Lehtimäki, T.; Hiltunen, R.

In: Journal of Chromatography B: Biomedical Sciences and Applications, Vol. 754, No. 2, 2001, p. 437-445.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Major human plasma lipid classes determined by quantitative high-performance liquid chromatography, their variation and associations with phospholipid fatty acids

AU - Seppänen-Laakso, Tuulikki

AU - Laakso, I.

AU - Vanhanen, H.

AU - Kiviranta, K.

AU - Lehtimäki, T.

AU - Hiltunen, R.

PY - 2001

Y1 - 2001

N2 - An HPLC method with evaporative light-scattering detection (ELSD) was optimized and validated for the simultaneous quantitation of cholesteryl esters (CEs), triacylglycerols (TGs), free cholesterol (FC) and phosphatidylcholine (PC) in human plasma. The separation of CEs from TGs, the most variable plasma lipid class, was improved by speeding up the gradient steps and by increasing the re-equilibration time between runs. The calibrations were made at levels of 0.14–14 μg lipid/injection. The intra- and inter-day precision values of the method ranged between 1.9 and 4.5 and 2.3–7.2% (RSD, n=6), respectively, including determinations at two concentration levels. In comparison to other lipid classes, quantitation of PC proved to be equally repeatable despite its lowest detector response. The relative recoveries varied from 97.0 to 110.3%, showing good accuracy of the method. The methodological variation of the lipid classes covered 0.6–3.1% of their total variation in the study population (n=48). The CE/FC ratio showed an even closer relationship with phospholipid linoleic acid (18:2n−6; r=0.65, P<0.001) than with serum cholesterol levels, while eicosapentaenoic acid (20:5n−3) was significantly associated with PC (r=0.41, P<0.01). The CE/FC ratio increased (P<0.01) during soyabean oil substitution and the level of PC increased (P<0.01) during cold-pressed rapeseed oil substitution.

AB - An HPLC method with evaporative light-scattering detection (ELSD) was optimized and validated for the simultaneous quantitation of cholesteryl esters (CEs), triacylglycerols (TGs), free cholesterol (FC) and phosphatidylcholine (PC) in human plasma. The separation of CEs from TGs, the most variable plasma lipid class, was improved by speeding up the gradient steps and by increasing the re-equilibration time between runs. The calibrations were made at levels of 0.14–14 μg lipid/injection. The intra- and inter-day precision values of the method ranged between 1.9 and 4.5 and 2.3–7.2% (RSD, n=6), respectively, including determinations at two concentration levels. In comparison to other lipid classes, quantitation of PC proved to be equally repeatable despite its lowest detector response. The relative recoveries varied from 97.0 to 110.3%, showing good accuracy of the method. The methodological variation of the lipid classes covered 0.6–3.1% of their total variation in the study population (n=48). The CE/FC ratio showed an even closer relationship with phospholipid linoleic acid (18:2n−6; r=0.65, P<0.001) than with serum cholesterol levels, while eicosapentaenoic acid (20:5n−3) was significantly associated with PC (r=0.41, P<0.01). The CE/FC ratio increased (P<0.01) during soyabean oil substitution and the level of PC increased (P<0.01) during cold-pressed rapeseed oil substitution.

KW - Cholesteryl esters

KW - Triacylglycerols

KW - Free cholesterol

KW - Phosphatidylcholine

KW - Phospholipid fatty acids

U2 - 10.1016/S0378-4347(01)00031-7

DO - 10.1016/S0378-4347(01)00031-7

M3 - Article

VL - 754

SP - 437

EP - 445

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

IS - 2

ER -