A miniaturized automated sample preparation method is described for the determination of fatty acid methyl esters in blood or plasma samples using the Agilent 7693 Series Automatic Liquid Sampler. Fifty-microliter samples are derivatized using acetyl chloride in methanol and the fatty acid methyl esters are extracted in iso-octane. Extracts are analyzed by GC-FID. Excellent repeatability is obtained and the method can be applied for fully automated fatty acid profiling in blood and plasma samples during dietary, biomedical or metabolomic studies.
The document details a fully automated sample preparation method for determining fatty acid methyl esters (FAMEs) in blood or plasma samples using the Agilent 7693 Series Automatic Liquid Sampler (ALS). The method involves derivatizing 50-microliter samples with acetyl chloride in methanol and extracting the fatty acid methyl esters in iso-octane for analysis by GC-FID.
The procedure utilizes a specific sequence programmed in GC ChemStation to handle reagents, heating, mixing, and extraction steps. The text provides a detailed sample preparation sequence, analytical conditions for the Agilent 7890 GC System, and results demonstrating excellent reproducibility with an average RSD of 2% for the analyzed fatty acids.
| Sample Volume | 50 microliters |
|---|---|
| Derivatization Reagent | 5% acetyl chloride in methanol |
| Extraction Solvent | iso-octane |
| Heating Temperature | 75 °C |
| Heating Duration | 600 seconds |
| Mixing Speed | 4000 rpm |
| Column Type | HP-88 |
| Inlet Temperature | 250 °C |
| Total Run Time | 84.33 min |
The method uses fifty-microliter samples of blood or plasma.
The sample is heated and mixed at 75°C for 600 seconds (10 minutes).
A long polar Agilent HP-88 GC column is used for optimal resolution.