Serum lipids
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Advances in Serum Lipid Determination and Analysis
Improved Colorimetric Methods for Serum Lipid Determination
Sulfo-Phospho-Vanillin Reaction
The sulfo-phospho-vanillin reaction has been a cornerstone in the colorimetric determination of total serum lipids. Recent improvements to this method have significantly enhanced its efficiency and accuracy. The modified procedure now requires only 20 µl of serum and is performed in a single tube, eliminating the need for tedious pipetting of a concentrated sulfuric acid-serum digest. This method demonstrates a linearity up to 1250 mg/dl and is unaffected by bilirubin concentrations up to 20 mg/dl, with a relative standard deviation of 4.9% for day-to-day measurements. Another study corroborates these findings, highlighting the method's simplicity, speed, and reliability, making it suitable for large-scale analyses with a coefficient of variation of 3.5%.
Enzymatic Summation Method
An alternative to colorimetric methods is the enzymatic summation method, which estimates total serum lipids by summing the concentrations of individual lipid components. This method has shown excellent correlation with traditional gravimetric analysis, with results agreeing within 1-3% across different serum samples. The enzymatic summation method is particularly advantageous for routine clinical use due to its reliability and convenience.
Quantitative Determination Techniques
Lipid Extraction and Gravimetric Analysis
Traditional methods for serum lipid determination, such as the Van Slyke and Plazin lipid extraction technique, involve coprecipitation of lipids and proteins, followed by extraction with methanol and chloroform. This method, while somewhat more variable than the Folch method, is simpler and faster, requiring less than half the performance time and no special apparatus.
High-Performance Liquid Chromatography (HPLC)
HPLC has been employed for the quantitative determination of fatty acid composition in serum lipids. This method uses fluorescence detection and has been validated against other quantitative analyses, showing reliable results. It is particularly useful for analyzing both free and total fatty acids in various patient populations, including normal subjects, diabetics, and those with angina pectoris.
Serum Lipidomics and Disease Biomarkers
Lipidomics Profiling Using LC-MS
Liquid chromatography-mass spectrometry (LC-MS) has become a powerful tool for lipidomics profiling, allowing for the qualitative and semiquantitative detection of lipids in biological samples. This method has been optimized for the detection and characterization of triglycerides (TGs) in serum, showing high accuracy and precision. It has been applied to study the effects of diet on serum TGs in rats, identifying 86 TG species that varied significantly with diet.
Surface-Assisted Laser Desorption Ionization Mass Spectrometry (SALDI-MS)
A novel SALDI-MS platform has been developed for high-coverage detection of serum lipids, capable of simultaneous detection of 236 lipid species in positive ion mode. This platform has been used to identify potential lipidomic biomarkers for hepatocellular carcinoma (HCC), achieving a diagnostic model with high sensitivity and specificity.
Serum Lipids and Inflammatory Bowel Disease (IBD)
Meta-Analysis of Serum Lipid Levels in IBD
A systematic review and meta-analysis have shown that serum lipid levels are significantly lower in IBD patients compared to healthy controls. Specifically, total cholesterol (TC), HDL-c, and LDL-c levels are reduced, with more pronounced decreases observed in Crohn's disease (CD) compared to ulcerative colitis (UC). The severity of IBD is negatively associated with serum lipid levels, indicating that lipid profiles could serve as biomarkers for disease activity.
Conclusion
The determination and analysis of serum lipids have seen significant advancements through improved colorimetric methods, enzymatic summation techniques, and sophisticated lipidomics profiling using LC-MS and SALDI-MS. These methods not only enhance the accuracy and efficiency of lipid measurement but also provide valuable insights into the role of lipids in various diseases, including IBD and HCC. As these techniques continue to evolve, they hold great promise for clinical diagnostics and personalized medicine.
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