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A rapid method for separation of plasma low and high density lipoproteins for tocopherol and carotenoid analyses

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Lipids

Abstract

Ultracentrifugation (UC) is the method most often employed for separation and quantification of lipoproteins. Because this procedure requires expensive laboratory equipment, a large volume of fresh sample and an inordinate amount of time, it may not be ideal for routine clinical/experimental use. The aim of the current study was to evaluate a method which combines selective precipitation (HDL-P) and immunoseparation (LDL-I) for the rapid and reliable isolation of high density lipoproteins (HDL) and low density lipoproteins (LDL) specifically for vitamin E and carotenoid determination within these fractions. Cholesterol and triacylglycerol concentrations within the HDL and LDL were also determined to enable expression of vitamin E and carotenoid concentrations per gram of lipid. Isolation of lipoproteins by UC was used as the reference method (HDL-UC/LDL-UC). There were no significant differences between methods for α-and γ-tocopherol in LDL and HDL. Carotenoids measured in HDL and LDL were comparable between the methods. The exception was higher lutein/zeaxanthin concentration in HDL-P and LDL-I compared to HDL-UC and LDL-UC, respectively. Additionally, lycopene concentration was significantly lower in LDL-I compared to LDL-UC. In comparing vitamin E and carotenoid values in lipoproteins separated from fresh and frozen plasma by the direct method, there was no difference in α-tocopherol or the majority of carotenoids measured. In conclusion, a combination of selective precipitation and immunoseparation of fresh or frozen plasma for subsequent α-and γ-tocopherol analyses provides an accurate and reliable alternative to lipoprotein separation by UC. Additionally, carotenoid concentrations in HDL separated by selective precipitation and analyses of α-and β-carotenes and β-cryptoxanthin in LDL separated by immunoseparation are also reliable, while lycopene and lutein/zeaxanthin concentrations in LDL-I are not readily comparable to LDL-UC.

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Abbreviations

HDL:

high density lipoproteins

HDL-P:

HDL isolated by selective precipitation

HDL-UC:

HDL isolated by sequential ultracentrifugation

HPLC:

high-performance liquid chromatography

LDL:

low density lipoprotein

LDL-I:

LDL isolated by immunoseparation

LDL-UC:

LDL isolated by sequential ultracentrifugation

TG:

triacylglycerol

UC:

ultracentrifugation

VLDL:

very low density lipoproteins

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Author notes

  1. John H. Contois

    Present address: Clinical Chemistry Laboratory, Hartford Hospital, 80 Seymour St., P.O. Box 5037, 06102-5037, Hartford, CT

Authors and Affiliations

  1. Department of Nutritional Sciences, University of Connecticut, 3624 Horsebarn Rd. Ext., 06269, Storrs, Connecticut

    Silke Vogel, John H. Contois, Sarah C. Couch & Carol J. Lammi-Keefe

Authors
  1. Silke Vogel
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  2. John H. Contois
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  3. Sarah C. Couch
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  4. Carol J. Lammi-Keefe
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Vogel, S., Contois, J.H., Couch, S.C. et al. A rapid method for separation of plasma low and high density lipoproteins for tocopherol and carotenoid analyses. Lipids 31, 421–426 (1996). https://doi.org/10.1007/BF02522929

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  • DOI: https://doi.org/10.1007/BF02522929

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