G005538

GCxGC Analysis of FAMEs in Human Plasma on Equity-1 x SUPELCOWAX 10, Non-Polar to Polar Strategy

suitable for GC

Properties

• technique(s): gas chromatography (GC): suitable
• test parameters: column: (¹D) Equity 1, 30 m x 0.25 mm I.D., 0.25 μm (28406-U); column: (²D) SUPELCOWAX 10, 0.95 m x 0.10 mm I.D., 0.10 μm (24343); oven: 180 °C, 3 °C/min to 280 °C (10 min); oven: (modulator) LMCS Everest longitudinally modulated cryogenic system, modulation period 6 sec; inj. temp.: 280 °C; detector: FID, 300 °C, 125Hz; MS, 20000 amu/sec; carrier gas: hydrogen; sample: human plasma
• application(s): clinical

Description

General description

The 0.95 m SUPELCOWAX 10 column was obtained by cutting lengths from a standard length column (e.g. 5, 10, 15 m).

Analysis Note

Sample prep: 100 mL of plasma was saponified with 1 mL of methanolic sodium methoxide (0.5% w/v) at a temperature of 100 °C for 15 min in a closed Pyrex tube. The subsequent methylic esterification was achieved with 1 mL boron trifluoride-methanol complex at a temperature of 100 °C for 15 min. The fatty acid methyl esters (FAMEs) were extracted by adding 1 mL of n-hexane to the mixture. One mL of a saturated NaCl solution was added to the entire mixture, which was then agitated manually for 2 min, followed by centrifugation at 3000 rpm for 10 min. After, anhydrous Na2SO4 was added to the Pyrex tube for drying and 0.9 mL of the n-hexane FAME layer was transferred to a GC injector vial. A 0.1 mL volume of 1000 ppm internal standard (C13:0) was also added to the GC vial.

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