Untargeted Lipidomics Service
Lipidomics (untargeted lipid profiling).
Lipids are first extracted using organic solvent and subjected to measurement using reversed phase chromatography coupled to high resolution Thermo Fisher mass spectrometer of Orbi trap type. For reduction of the complexity of extracted lipids, a column with C8 stationary phase is used to separate the lipids and other lipophilic compounds based on their hydrophobicity. The differences in elution time enable detection and relative quantitation of bigger number of lipids of the same head groups or carbon chain.
Untargeted Lipidomics services – detectable lipid classes
Free Fatty acids
FA
Diacylglycerol
DAG
Triacylglycerol
TAG
Phosphatidylcholine
PC
Phosphatidylinositol
PI
Phosphatidylethanolamine
PE
Phosphtidylglycerol
PG
Lysophosphatidylcholine
LysoPC
Lysophosphatidylethanolamine
LysoPE
Lysophosphatidylglycerol
LysoPG
Ether-linked phosphatidylcholine
ePC
Ether-linked Phosphatidylethanolamine
ePE
Ether-linked Lysophosphatidylcholine
eLysoPC
Sphingomyelins
SM
Ceramides
Cer
Hexosylceramide
HexCer
Inositol phosphorylceramide
IPC
Cardiolipins
CL
Sterol Esters
SE
Acyl carnitines
AC
Monogalactosyldiacylglycerols
MGDG
Digalactosyldiacylglycerols
DGDG
Sulfoquinovosyl diacylglycerols
SQDG
Cholic acid, Sphingosine
Liquid chromatography (reversed phase) - Mass spectrometry-based untargeted lipids profiling
Analysis of lipids and lipophilic compounds in complex samples using high resolution mass spectrometers in untargeted manner results in detection of 1000-3000 features of which up to 600 and 550 can be identified (annotated) in animals' tissues and human serum respectively.
23 different lipids classes and carnitines can be annotated with use of internal database and LC-MS/MS annotation algorithm. They are represented by different number of lipids different sample types.
Annotation is performed using metaSysX database which contains information 1400 lipids. Samples are additionally measured in LC-MS/MS positive and negative ionization modes and are subjected to annotation using a fragmentation spectrum which increases the annotation level and results in the highest coverage of annotated lipids.
Measurements are performed in positive and negative ionization mode to increase the number of annotated compounds.
Untargeted lipidomic analysis results in a combination of annotated lipids and unknown features characterized by mass-to charge ratio.
Our annotation analysis results in following information about each lipid detected in the data:
- LIPID HEADGROUPS for glycerophospholipids, sphingolipids glycolipids and sterols
- FATTY ACIDS CHAIN AND NUMBER OF DOUBLE BONDS
Example of annotation analysis:
PC 34:0
PC 34:1
PC:34:2
LysoPC 20:2
DAG 34:1
SM 28:1
Cer(d16:1/24:1)
SM 31:1
Cer(d18:1/25:1)
18:3-Cholesterol
Cer(d18:1/25:0)
GlcCer(t18:0/24:1)
GlcCer(d18:1/24:0(2OH))
TAG 49:5
SM 43:1
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