Lipids are biomolecules that are soluble in non-polar solvents, including fats, sterols, fat-soluble vitamins (such as vitamins A, D, E, and K), monoglycerides, diglycerides, triglycerides, and sheaths Lipids and phospholipids and so on. It is the main component that constitutes the biomembrane, and regulates the signal transduction process of cell growth, differentiation, senescence, and programmed death. It is also involved in physiological functions such as body growth, health, intellectual development, and memory. In addition, it can also affect the occurrence and development of various diseases such as tumor, cardiovascular, endocrine, immune, and nervous system.
When you want to study a specific target lipid or lipid class, you can choose targeted lipidomics. Precise method development allows for the inclusion of equal amounts of isotope-labeled or odd-chain internal standards selected from customizable lipid panels. Lipids from each sample are extracted using standardized protocols and analyzed by GC/LC-MS/MS. Analyte concentrations are determined from calibration curves prepared from authentic standards. This approach is especially useful for quantifying lipids present at low levels in samples, such as endogenous bioactive lipids (e.g., prostaglandins, leukotrienes, or endocannabinoids). Standard statistical analysis tools can reveal changing trends in lipid species among experimental groups, which can be visualized with a heat map or other available graphical tools.
Creative Proteomics provides reliable, fast, and cost-effective targeted lipidomics to accelerate your project.
Liquid chromatography or gas chromatography hyphenated to triple quadrupole mass spectrometers for targeted lipidomics.
Fig1. The protocol workflow of targeted lipidomics (Creative Proteomics)
|Sample Type||Sample Collection||Lysis Method||Recommended Quantity||Additional Considerations|
|Tissues||Homogenization for cell release||Tissue-specific lysis protocols||10-50 mg tissue||Tissue-specific considerations; avoid contamination during handling.|
|Cell Culture||Harvesting with trypsin or other methods||Specialized buffers for efficient lysis||Varies based on culture dish||Consistent culture conditions and avoiding contamination are crucial.|
|Plasma/Serum||Blood collection with anticoagulants||Protein precipitation or organic extraction||100-500 μL||Immediate sample processing to prevent lipid alterations.|
|Cell Pellets||Centrifugation and washing||Specialized detergents for efficient lysis||1-5 x 10^6 cells||Optimize washing steps to minimize cell residue.|
|Lipoproteins||Ultracentrifugation or density gradient||Organic extraction or specialized methods||50-200 μL||Handle lipoproteins with care to prevent structural alterations.|
|Exosomes||Ultracentrifugation or precipitation||Specialized extraction methods||100-500 μL||Maintain proper exosome isolation techniques for accurate analysis.|
|Breast Milk||Collection in clean containers||Lipid extraction from milk||5-10 mL||Minimize contamination during collection and processing.|
|Skin Biopsy||Biopsy procedure||Tissue-specific lysis protocols||5-10 mg tissue||Preserve sample integrity and prevent degradation.|
|Urine||Midstream collection in sterile containers||Organic extraction or precipitation||5-10 mL||Process samples promptly to prevent lipid changes.|
|Feces||Fresh collection in airtight containers||Homogenization and organic extraction||100-500 mg||Minimize oxygen exposure to maintain sample stability.|
|Plants||Harvest and flash freeze||Tissue-specific lysis protocols||50-200 mg||Rapid freezing preserves lipid profiles in plant tissues.|
|Microorganisms||Culture or direct collection||Cell lysis and extraction methods||Varies based on biomass||Choose appropriate lysis methods for different microorganisms.|
|Food||Homogenization or extraction||Solvent-based extraction methods||Varies based on food type||Tailor extraction methods to the characteristics of the food sample.|
If you have any questions about our targeted lipidomics services, please contact us.
Case: PCB153-Mediated Disruption of Glycerophospholipid Metabolism in PC12 Cells: A Lipidomics Study
This study focuses on understanding the toxicological effects of PCB153 on glycerophospholipid metabolism in PC12 cells. PCB153, a polychlorinated biphenyl, is known to have adverse health effects, and this research aims to unravel its specific influence on lipid metabolism.
PC12 cells were utilized in the study, obtained from the Chinese Academy of Sciences, Shanghai, China. The exposure to PCB153 occurred at three different concentrations (0.05 μg mL−1, 0.5 μg mL−1, and 20 μg mL−1) over a period of 120 hours.
Chemicals: High-quality chemicals were employed, including HPLC-MS grade acetonitrile and methanol, HPLC grade 2-propanol and dichloromethane, and HPLC-MS grade ammonium acetate.
Cell Culture: PC12 cells were cultured in RPMI 1640 medium supplemented with fetal bovine serum, penicillin, and streptomycin, maintained at 37 °C with 5% CO2.
Cell Viability Assay: PC12 cells were exposed to different PCB153 concentrations, and viability was measured using the cell counting kit-8 (CCK-8) method at 24-hour intervals up to 120 hours.
Sample Collection and Preparation: Cell samples were collected at various time points, and glycerophospholipids were extracted using a modified method. UHPLC-MS/MS was employed for lipid analysis, and data analysis included statistical methods such as t-tests, PCA, and OPLS-DA.
Method Validation: The study confirmed the presence of 22 glycerophospholipids in PC12 cells. The extraction method was validated for efficiency and stability.
PCB153 Effects on PC12 Cell Viability: Exposure to PCB153 resulted in dosage-dependent decreases in cell viability. Multivariate statistical analysis highlighted the comprehensive impact of PCB153 on lipid metabolism.
Identification of Potential Biomarkers: Five potential glycerophospholipid biomarkers (PC(14:0/14:0), PE(16:0/18:1), PE(16:0/18:2), PS(18:0/18:1), PI(16:0/18:1)) were identified, suggesting their roles in cellular functions and potential implications for neurodegenerative disorders.
Implications and Conclusion: The study suggests a link between PCB153 exposure and glycerophospholipid quantity changes, emphasizing the importance of metabolic disturbances in understanding the toxicity mechanisms. Lipidomics studies could provide valuable insights for risk assessment and neuroprotective therapeutic development.
The OPLS-DA results among control and dosage groups from 24 h to 120 h
Heat maps generated by Hierarchical Pearson clustering for 22 target glycerophospholipids after exposure of PCB153 at 96 h (A), 120 h (B)