The lipids extracted from LM were methylated according to the method by Lepage and Roy (1986) (link) and the fatty acid composition was analyzed using gas chromatography. Approximately 200 μL of each lipid extract were converted into fatty acid methyl esters by adding 2 mL methanol:benzene (4:1, vol:vol) solution and 200 μL of acetyl chloride, and heated for 40 min at 100°C on a heating block (Lab & Tools, Anyang, Korea). One mL isooctane (Sigma-Aldrich, Saint Louis, MO, USA) and 8 mL of 6% potassium carbonate (Junsei, Tokyo, Japan) were added, and the solution was centrifuged (1580R, Labogene, Daejeon, Korea) for 10 min at 500×g. One (1) μL of the supernatant was subjected to analysis with a gas chromatography (Clarus 500; Perkin Elmer, Shelton, CT, USA) equipped with a fused column SP 2560 (100 m in length, 0.25 mm in diameter, 0.2 um film thickness) using nitrogen as the carrier gas (flow rate of 1.0 mL/min). The initial oven temperature was 210°C; the injector and detector temperatures were 240°C and 250°C, respectively.
Sp 2560
Manufactured by PerkinElmer
Sourced in United States
The SP 2560 is a high-performance gas chromatography (GC) column designed for the separation and analysis of fatty acid methyl esters (FAMEs). It features a highly polar stationary phase that provides efficient separation of a wide range of FAME compounds, enabling accurate quantification and identification.
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Lab products found in correlation
4 protocols using sp 2560
1
Fatty Acid Composition Analysis
2
Fatty Acid Profiling of Lipid Extracts
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The extracted lipids from LM were acetylated by using the method of Lepage and Roy [11 (link)], and fatty acid composition was analyzed by using gas chromatography. Approximately 100 μL of lipid extract were converted to fatty acid methyl esters by adding 2 mL methanol:benzene (4:1, v/v) and 200 μL of acetyl chloride and heating for 40 minutes on a heating block. Isooctane (1 mL) and 6% potassium carbonate (8 mL) were added the mixture and centrifuged for 10 minutes at 1,500×g. The supernatant was used to analyze fatty acid composition by using a gas chromatograph (Clarus 500, Perkin Elmer, Shelton, CT, USA) equipped with a fused column SP 2560 (100 m length, 0.25 mm diameter) and nitrogen as the carrier gas (flow rate; 1 mL/min). The initial oven temperature was 200°C and the injector and detector temperatures were 220°C and 250°C, respectively.
3
Fatty Acid Profiling via GC-FID
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Fatty acids methyl esters (FAME) were prepared using boron trifluoride/methanol [22 ], and lipid profiles were determined through gas chromatography using a Perkin Elmer Clarus 500 with a flame ionization detector (FID) and a capillary column SP-2560 (100 m × 0.25 mm × 0.2 µm). The injector and detector temperatures were set at 250 and 260 °C, respectively. The oven temperature was set at 175 °C for 10 min, increased to 200 °C at a rate of 10 °C/min, and then increased to 220 °C at a rate of 4 °C/min and held for 15 min. Hydrogen was used as the carrier gas, with a pressure of 30 psi and a split ratio of 120 mL/min. The identification and quantification of the fatty methyl esters were carried out using heptadecanoic acid methyl ester as an internal standard [23 (link)].
4
Fatty Acid Composition Analysis of Freeze-Dried LD Muscle
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A total of 0.50 g freeze-dried LD muscle samples were accurately weighed to analyze the fatty acid composition according to the method described by Folch et al. [21 (link)] using gas chromatography equipped with flame ionization detection (Clarus680, PerkinElmer, Inc., Waltham, MA, USA) using an SP2560 capillary column (100 m × 0.25 mm × 0.2 μm). Methyl nonadecanoate was used as an internal standard. The injector and detector temperatures were maintained at 220 °C and 260 °C, respectively. The initial oven temperature was programmed at 120 °C for 5 min, and the temperature was then increased to 230 °C at 3 °C/min, held for 5 min, increased to 240 °C at 1.5 °C/min, and held for 13 min, with a split ratio of 1:10. Nitrogen was used as the carrier gas with a flow rate of 1 mL/min.
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