Neutral Lipid Analysis

Neutral Lipids: are hydrophobic molecules that lack charged groups. The group of neutral lipids consists of triglycerides, steryl esters and wax esters. Triglycerides are present in all cell types and serve as storage molecules for free fatty acids, diglycerides and monoglycerides, which are then utilized as energy sources. Triglycerides are used as a substrate for the synthesis of these molecules and their degradation results in the formation of mono- and diglycerides.

Triglycerides: consist of three fatty acid chains bound to a glycerol molecule via ester linkages. Triglycerides function as intracellular storage molecules that are later broken down to provide energy for the cell. They are the most common form of fat in the body and the main component in vegetable oils and animal fat. High levels of triglycerides have been associated with increased risk of atherosclerosis, fatty lover disease and pancreatitis.

Diglycerides: consist of two fatty acid chains bound to a glycerol molecule via ester linkages. They are involved in cell membrane properties, signalling, cell metabolism, etc. Diglycerides are common components of edible oils and are often used as food emulsifiers, stabilizers and thickeners. They are present in commercially-produced seed oils and can affect the physical properties of these products.

Monoglycerides: consist of a single fatty chain bound to a glycerol molecule via an ester linkage. Monoglycerides are also commonly used as food emulsifiers and occur naturally in many food products. Both mono- and diglycerides are classified as “generally recognized as safe” in small amounts by the U.S. Food and Drug Administration.

Analytical Targets – triglycerides, diglycerides, monoglycerides.

Neutral Lipid Sources - Common sources of industrially produced neutral lipids include edible oils, nutraceuticals oils, food products, biological material (e.g. whole blood, red blood cells), animal fats, etc. Each source has a unique profile of individual neutral lipids and consequently differing applications in food, nutrition, pharmaceuticals, cosmetics and drug delivery.

Analysis Platform - GC-FID

Method Summary

Lipids including Free Fatty Acids (FFA) are extracted from samples according to the method of Bligh & Dyer (1) in the presence of an internal standard. Subsequent processing using methods similar to those described (1). Extracts were then separated by solid-phase extraction by spotting the lipid extracts containing internal standards on silica gel. The thin layer chromatography plate is developed in the appropriate mobile phase for the target analyte. The plate is then dried and sprayed with ANS in order to identify bands of interest. The bands are identified by RF comparison to appropriate standards by using long wave UV. Bands of interest are removed, and their fatty acid methyl esters are transmethylated. The resulting fatty acid methyl esters are then analyzed on a Agilent 7890B gas-liquid chromatograph with a 60-m DB-23 capillary column (0.32 mm internal diameter) against their internal standard. Data is reported in ug/g and % by weight.

Sample Requirement

  • Liquid Medium – Oils, Biological Cells (RBC / WBC / Serum)– minimum 2.5g equivalency per analysis.
  • Tissues/foods/powders – Grains, powders, foods – minimum of 2.5g equivalency per analysis.
  • Please contact with regards to your specific medium.

Data Reporting

  • CoA (pdf.) provided for sample analysis.
  • Data reported as: μg/g and % by weight.
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