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Development of Enhanced Analytical Methodology for Lipid Analysis from Sampling to Detection: A Targeted Lipidomics Approach
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Department of Chemistry, Analytical Chemistry.
2005 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis covers a wide range of analytical method development for lipid analysis in complex biological samples; from sample preparation using pressurized fluid extraction (PFE) and separation with reversed phase capillary liquid chromatography (RP-LC) to detection by electrospray ionization mass spectrometry (ESI/MS) and tandem MS.

The requirements for fast, reliable and selective extraction methods with minimal usage of solvents have accelerated the development of new extraction techniques. PFE is one of the new automated, fast and efficient liquid extraction techniques which use elevated temperature and pressure with standard liquid solvents. In this thesis the reliability and efficiency of the PFE technique was investigated for the extraction of total lipid content from cod, herring muscle and human brain tissue as well as for pesticides from fatty foodstuffs. Improved or comparable efficiencies were achieved with reduced time and solvent consumption as compared to traditional methods.

A RP-LC coupled online to ESI/MS for the analysis of phosphatidylcholine (PC) and sphingomyelin (SM) molecular species was developed and used for the analysis of brain lipids from eight groups of mice treated with vehicle and various neuroleptics. The effect of postnatal iron administration in lipid composition and behavior was investigated. Whether or not these effects could be altered by subchronic administration of the neuroleptics (clozapine and haloperidol) were examined. The results support the hypothesis that an association between psychiatric disorders, behavior abnormalities and lipid membrane constitution in the brain exists.

Finally, a tandem MS precursor ion scan was used to analyze the developmental profile of brain sulfatide accumulation in arylsulfatase A (ASA) deficient (ASA -/-) as compared to wild type control (ASA +/+) mice. The ASA -/- mice were developed as a model of the monogenic disease metachromatic leukodystrophy with an established deficiency of the lysosomal enzyme ASA. The results showed that an alteration in the composition of sulfatide molecular species was observed between the ASA -/- and ASA +/+ mice.

This thesis shows that modern analytical methods can provide new insights in the extraction and analysis of lipids from complex biological samples.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis , 2005. , p. 48
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 56
Keyword [en]
Analytical chemistry, electrospray ionization, lipids, liquid chromatography, mass spectrometry, pesticides, phosphatidylcholine, pressurized fluid extraction, sphingomyelin, sulfatides
Keyword [sv]
Analytisk kemi
National Category
Analytical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-5810ISBN: 91-554-6259-6 (print)OAI: oai:DiVA.org:uu-5810DiVA, id: diva2:166474
Public defence
2005-05-20, Room B41, Biomedical Center (BMC), Uppsala, 13:15
Opponent
Supervisors
Available from: 2005-04-29 Created: 2005-04-29Bibliographically approved
List of papers
1. Total Lipid Extraction of Homogenized and Intact Lean Fish Muscles Using Pressurized Fluid Extraction and Batch Extraction Technique
Open this publication in new window or tab >>Total Lipid Extraction of Homogenized and Intact Lean Fish Muscles Using Pressurized Fluid Extraction and Batch Extraction Technique
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2005 (English)In: Journal of Agricultural and Food Chemistry, ISSN 0021-8561, E-ISSN 1520-5118, Vol. 53, no 14, p. 5506-5512Article in journal (Refereed) Published
Abstract [en]

The reliability and efficiency of pressurized fluid extraction (PFE) technique for the extraction of total lipid content from cod and the effect of sample treatment on the extraction efficiency have been evaluated. The results were compared with two liquid-liquid extraction methods, traditional and modified methods according to Jensen. Optimum conditions were found to be with 2-propanol/n-hexane (65:35, v/v) as a first and n-hexane/diethyl ether (90:10, v/v) as a second solvent, 115 oC, and 10 min of static time. PFE extracts were cleaned up using the same procedure as in the methods according to Jensen. When total lipid yields obtained from homogenized cod muscle using PFE were compared yields obtained with original and modified Jensen methods, PFE gave significantly higher yields, ~10% higher (t test, P < 0.05). Infrared and NMR spectroscopy suggested that the additional material that inflates the gravimetric results is rather homogeneus and is primarily consists of phospholipid with headgroups of inositidic and/or glycosidic nature. The comparative study demonstrated that PFE is an alternative suitable technique to extract total lipid content from homogenized cod (lean fish) and herring (fat fish) muscle showing a precision comparable to that obtained with the traditional and modified Jensen methods. Despite the necessary cleanup step, PFE showed important advantages in the solvent consumption was cut by ~50% and automated extraction was possible.

Keyword
Lean fish, phospholipids, pressurized fluid extraction, PFE, total lipid content
National Category
Natural Sciences
Identifiers
urn:nbn:se:uu:diva-93095 (URN)10.1021/jf0501286 (DOI)15998106 (PubMedID)
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2017-12-14Bibliographically approved
2. Pressurized Fluid Extraction (PFE) as an Alternative General Method for the Determination of Pesticide Residues in Rape Seed
Open this publication in new window or tab >>Pressurized Fluid Extraction (PFE) as an Alternative General Method for the Determination of Pesticide Residues in Rape Seed
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2002 In: The Analyst, ISSN 0003-2654, Vol. 127, no 4, p. 554-559Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-93096 (URN)
Available from: 2005-04-29 Created: 2005-04-29Bibliographically approved
3. Analysis of Phosphatidylcholine and Sphingomyelin Molecular Species From Brain Extracts Using Capillary Liquid Chromatography Electrospray Ionization Mass Spectrometry
Open this publication in new window or tab >>Analysis of Phosphatidylcholine and Sphingomyelin Molecular Species From Brain Extracts Using Capillary Liquid Chromatography Electrospray Ionization Mass Spectrometry
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2003 In: Journal of Neuroscience Methods, ISSN 0165-0270, Vol. 128, no 1-2, p. 111-119Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-93097 (URN)
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2011-03-21
4. Brain Lipid Composition in Postnatal Iron-Induced Motor Behavior Alterations Following Chronic Neuroleptic Administration in Mice
Open this publication in new window or tab >>Brain Lipid Composition in Postnatal Iron-Induced Motor Behavior Alterations Following Chronic Neuroleptic Administration in Mice
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In: Neuroscience LettersArticle in journal (Refereed) Submitted
Identifiers
urn:nbn:se:uu:diva-93098 (URN)
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2011-03-21
5. Characterization of Brain Sulfatide Molecular Species in Arylsulfatase A Deficient Mice Using Electrospray Ionization Tandem Mass Spectrometry
Open this publication in new window or tab >>Characterization of Brain Sulfatide Molecular Species in Arylsulfatase A Deficient Mice Using Electrospray Ionization Tandem Mass Spectrometry
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Manuscript (Other academic)
Identifiers
urn:nbn:se:uu:diva-93099 (URN)
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2011-03-21

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