Identification of Lipids in Bovine Brains Using Untargeted Liquid Chromatography and Mass Spectrometry
Disciplines
Analytical Chemistry | Biochemistry
Abstract (300 words maximum)
Lipids are a crucial molecule for maintaining healthy brain cells. Imbalances of lipid classes are strongly correlated with certain diseases of the brain, which can extend to the rest of the body. Knowing the general lipid composition of healthy mammalian brains helps to identify changes that are caused by disease and use those markers to help develop more accurate diagnostic tests or even therapeutics for the disease. In this study, the lipid composition of bovine brain was determined by utilizing untargeted Liquid Chromatography-Mass Spectrometry (LCMS). This technique allows for accurate identification of the lipid classes found in a brain sample that has been prepared using Folch extraction. The Folch extraction is performed using a mixture of methanol and chloroform mixed with a brain sample. The mixture is allowed to incubate at room temperature for an hour while being mixed by a shaker to help extract the lipids. Then, water is added to mixture and centrifuged to separate the organic phase and the aqueous phase. The organic phase is transferred to another microcentrifuge tube to evaporate the organic solvent using a vacuum centrifuge. The sample is then reconstituted in methanol: acetonitrile (90:10). Once reconstituted, lipids are separated using a reverse phase C18 column in a Vanquish Flex HPLC, with a 12-minute gradient containing water and acetonitrile. The mass spectrometer is coupled with the LC in positive and negative mode to determine the m/z of the lipids present in the sample. Initially Compound Discover software was used to identify preliminary matches which then further filtered using the LIPIDMAPS software to classify the lipids such as Sphingolipids, Glycerolipids, Fatty acyls, Glycerophospholipids, and Sterol lipids. Preliminary analysis of a bovine brain showed the most prevalent lipid classes to be fatty acyls, by a large margin, followed by Glycerophospholipids.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Identification of Lipids in Bovine Brains Using Untargeted Liquid Chromatography and Mass Spectrometry
Lipids are a crucial molecule for maintaining healthy brain cells. Imbalances of lipid classes are strongly correlated with certain diseases of the brain, which can extend to the rest of the body. Knowing the general lipid composition of healthy mammalian brains helps to identify changes that are caused by disease and use those markers to help develop more accurate diagnostic tests or even therapeutics for the disease. In this study, the lipid composition of bovine brain was determined by utilizing untargeted Liquid Chromatography-Mass Spectrometry (LCMS). This technique allows for accurate identification of the lipid classes found in a brain sample that has been prepared using Folch extraction. The Folch extraction is performed using a mixture of methanol and chloroform mixed with a brain sample. The mixture is allowed to incubate at room temperature for an hour while being mixed by a shaker to help extract the lipids. Then, water is added to mixture and centrifuged to separate the organic phase and the aqueous phase. The organic phase is transferred to another microcentrifuge tube to evaporate the organic solvent using a vacuum centrifuge. The sample is then reconstituted in methanol: acetonitrile (90:10). Once reconstituted, lipids are separated using a reverse phase C18 column in a Vanquish Flex HPLC, with a 12-minute gradient containing water and acetonitrile. The mass spectrometer is coupled with the LC in positive and negative mode to determine the m/z of the lipids present in the sample. Initially Compound Discover software was used to identify preliminary matches which then further filtered using the LIPIDMAPS software to classify the lipids such as Sphingolipids, Glycerolipids, Fatty acyls, Glycerophospholipids, and Sterol lipids. Preliminary analysis of a bovine brain showed the most prevalent lipid classes to be fatty acyls, by a large margin, followed by Glycerophospholipids.