Free fatty acids (FFA) are an important part of lipid metabolism. In animals, many dietary lipids are hydrolyzed into free fatty acids before being absorbed and further used for lipid synthesis. Lipids can be hydrolyzed into free fatty acids by lipolytic enzymes, such as hormone-sensitive lipase, lipoprotein lipase, and phospholipase A and C, and then metabolized in many different ways.
As the main energy fuel for cell energy, different types of fatty acids are related to various metabolic diseases such as diabetes, coronary heart disease and non-alcoholic fatty liver. Free fatty acid analysis can be used as a potential biomarker for disease assessment. The detection of free fatty acid levels can also reflect the body's lipid metabolism, sugar metabolism and endocrine function status. In addition, the analysis of various subtypes of fatty acids (such as omega-3 and omega-6) in food ingredients and dietary supplements (such as fish oil) also has research and commercial significance.
In biological samples, free fatty acids account for only 10% of the total fatty acids, with low content and a wide dynamic range. Creative Proteomics has developed a free fatty acid analysis platform that can simultaneously detect and quantify as many free fatty acids as possible in biological samples at one time. Based on LC-MS/MS technology, it can qualitatively and quantitatively detect a variety of biological samples, including urine, whole blood/serum/plasma, etc.
Creative Proteomics use negative ion chemical ionization gas chromatography hyphenated to triple quadrupole mass spectrometers for free fatty acids analysis.
Fig1. The workflow of free fatty acids analysis service.
|Cells||Cell number should be kept below 2 million (common use is 0.5 million cells). 100 uL Internal Standard is added. Two volumes methanol lyses the cells and the mixture is acidified with HCl to 25 mM final concentration.|
|Media||100uL internal standard is added to 0.5 ml media. Then samples is mixed with 1 volume methanol and acidified with HCl to 25 mM final concentration.|
|Plasma||For blood plasma, 200uL is added to 300uL dPBS. Then 100uL Internal Standard is added and mixed with 1 volume methanol and acidified with HCL to 25 mM final concentration.|
|Tissue||For Tissue, the amount used should be empirically determined beforehand based on tissue type used. Add 100uL Internal Standard. Then add 900uL methanol. For tougher Tissues, samples are ground/minced, then sonicated. For softer tissues, just sonication is required. Sample is then mixed.|
If you have any questions about our free fatty acids analysis services, please contact us.