LC-MS-based method for the qualitative and quantitative analysis of complex lipid mixtures

Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, Boston, MA, USA.
The Journal of Lipid Research (Impact Factor: 4.42). 05/2006; 47(4):804-14. DOI: 10.1194/jlr.M500506-JLR200
Source: PubMed


A simple and robust LC-MS-based methodology for the investigation of lipid mixtures is described, and its application to the analysis of human lipoprotein-associated lipids is demonstrated. After an optional initial fractionation on Silica 60, normal-phase HPLC-MS on a YMC PVA-Sil column is used first for class separation, followed by reversed-phase LC-MS or LC-tandem mass spectrometry using an Atlantis dC18 capillary column, and/or nanospray MS, to fully characterize the individual lipids. The methodology is applied here for the analysis of human apolipoprotein B-associated lipids. This approach allows for the determination of even low percentages of lipids of each molecular species and showed clear differences between lipids associated with apolipoprotein B-100-LDL isolated from a normal individual and those associated with a truncated version, apolipoprotein B-67-containing lipoproteins, isolated from a homozygote patient with familial hypobetalipoproteinemia. The methods described should be easily adaptable to most modern MS instrumentation.

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    • "The ionization conditions can be further improved (matrix effects reduced) by carefully purifying the sample prior to LC–MS/MS. LC–MS/MS may yield better sensitivity and detection of minor components than the shotgun approach due to the sequential ionization of the analytes and may reduce the matrix background and lead to a more accurate determination of the noise and ionic background [8] [9] [12]. "
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    ABSTRACT: A new, sensitive and selective liquid chromatography-electrospray ionization-tandem mass spectrometric (LC-ESI-MS/MS) method was developed for the analysis of Phospholipids (PLs) in bio-oils and fats. This analysis employs hydrophilic interaction liquid chromatography-scheduled multiple reaction monitoring (HILIC-sMRM) with a ZIC-cHILIC column. Eight PL class selective internal standards (homologs) were used for the semi-quantification of 14 PL classes for the first time. More than 400 scheduled MRMs were used for the measurement of PLs with a run time of 34min. The method's performance was evaluated for vegetable oil, animal fat and algae oil. The averaged within-run precision and between-run precision were ≤10% for all of the PL classes that had a direct homologue as an internal standard. The method accuracy was generally within 80-120% for the tested PL analytes in all three sample matrices. Copyright © 2015 Elsevier B.V. All rights reserved.
    Journal of Chromatography B 08/2015; 1001:140-149. DOI:10.1016/j.jchromb.2015.07.036 · 2.73 Impact Factor
    • "Advantages of shotgun are an easier automation and higher throughput, while HPLC/MS is more prone to ion suppression effects and can provide more detailed information of various types of lipid isomerism. HILIC can be used for the separation of individual classes of polar lipids [24] [25] [26] [27] [28], while NP-HPLC is more convenient for nonpolar lipid classes [29] [30] [31] [32]. The lipid species separation according to the acyl chain length and the number of the double bonds can be achieved by reversed phase (RP)-HPLC [25,33–36], while silver-ion HPLC is more convenient for nonpolar lipid regioisomers or double bond positional isomers [37] [38]. "
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    ABSTRACT: Differences among lipidomic profiles of healthy volunteers, obese people and three groups of cardiovascular disease (CVD) patients are investigated with the goal to differentiate individual groups based on the multivariate data analysis (MDA) of lipidomic data from plasma, erythrocytes and lipoprotein fractions of more than 50 subjects. Hydrophilic interaction liquid chromatography on ultrahigh-performance liquid chromatography (HILIC-UHPLC) column coupled with electrospray ionization mass spectrometry (ESI-MS) is used for the quantitation of four classes of polar lipids (phosphatidylethanolamines, phosphatidylcholines, sphingomyelins and lysophosphatidylcholines), normal-phase UHPLC-atmospheric pressure chemical ionization MS (NP-UHPLC/APCI-MS) is applied for the quantitation of five classes of nonpolar lipids (cholesteryl esters, triacylglycerols, sterols, 1,3-diacylglycerols and 1,2-diacylglycerols) and the potential of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is tested for the fast screening of all lipids without a chromatographic separation. Obtained results are processed by unsupervised (principal component analysis) and supervised (orthogonal partial least squares) MDA approaches to highlight the largest differences among individual groups and to identify lipid molecules with the highest impact on the group differentiation. Copyright © 2015 Elsevier B.V. All rights reserved.
    Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 05/2015; 990. DOI:10.1016/j.jchromb.2015.03.010 · 2.73 Impact Factor
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    • "The lipids in each class were quantified with an internal standard of that class. A simple and robust LC-MS-based methodology using external standards for semiquantitative characterization of the total lipids was also described by Sommer et al. (2006). In our work, the dynamic range/linearity of each lipid class was determined with mixed standard lipids up to 5 nmol mL −1 . "
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    ABSTRACT: A sensitive method based on electrospray ionization tandem mass spectrometry was used to profile glycerolipids in Pyropia haitanensis and their changes responding to agaro-oligosaccharides. Ten monogalactosyldiacylglycerols (MGDGs), twelve digalactosyldiacylglycerols (DGDGs), five sulfoquinovosyldiacylglycerols (SQDGs), five phosphatidylglycerols (PGs), fifteen phosphatidylcholins (PCs), three phosphatidic acids (PAs), and three phosphatidylethanolamines (PEs) were identified in P. haitanensis. We found the SQDG was the most abundant species, followed by MGDG, DGDG, PG, PC, PE, and PA of the total glycerolipids. The predominant lipid species contained C20 fatty acids at sn-1/sn-2 positions, which was different from higher plants. Changes in membrane lipid species occurred when P. haitanensis were treated with agaro-oligosaccharides. At first, agaro-oligosaccharides induced an increase in total glycerolipids including the galactolipids such as MGDG (20:5/20:5) and phospholipids such as PC (18:3/20:5), suggesting that agaro-oligosaccharides caused changes of lipids in chloroplasts and plasma membrane. With increased treatment time, a large decline in major plasma membrane lipids (PCs and PEs) was observed, but not galactolipids (MGDGs and DGDGs), suggesting that the lipid changes occurred mainly at the plasma membrane after prolonged treatment.
    Journal of Applied Phycology 12/2014; 26(3):2397-2404. DOI:10.1007/s10811-014-0276-8 · 2.56 Impact Factor
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