The fatty acids of extract from C. obtusa were analyzed by gas chromatography. Using HP 5890 system (HP 19251A, Hewlett-Packard, Palo Alto, CA, USA) was used and flame ionization detector was used. The BF3 method (Dawidowicz and Thompson, 1971 (link)) was modified and the fatty acid standard (Sigma, St. Louis, MO, USA), used in the methylation, was C14, C16, C18, C18:1, C18:2. The extracts used 2N-KOH/MeOH for methylation. The column used Supelco SP-2340 Fused Silica Capillary Column (30 m, 0.25 mm [in side diameter], 0.20 μm [Film]) and the oven condition was 170°C→2°C/min→220°C 2 min hold, the detector and the injector temperature set at 230°C.
Sp 2340 fused silica capillary column
Manufactured by Merck Group
Sourced in United States
The SP-2340 fused silica capillary column is a gas chromatography (GC) column designed for the separation and analysis of various chemical compounds. It is made of fused silica, which provides high thermal stability and inertness. The column's core function is to facilitate the separation and identification of analytes in complex samples through GC analysis.
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4 protocols using sp 2340 fused silica capillary column
1
Fatty Acid Profiling of C. obtusa
2
Fatty Acid Composition Analysis by GC
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The fatty acid composition was determined after sample transesterification with KOH 2N in methanol [23 ,31 (link)] using a gas-chromatograph system composed of an Agilent Technologies 7890 (Agilent Technologies Inc., Santa Clara, CA, USA), equipped with an FID detector (set at 220 °C) and an SP™ 2340 fused silica capillary column (Supelco, Bellefonte, PA, USA), 60 m length × 0.25 mm i.d. and 0.20 μm film thickness. The temperature of the split injector was 210 °C, with a splitting ratio of 1:100; the detector temperature was 220 °C. The oven temperature was programmed as follows: at the very beginning, the temperature was set at 160 °C then gradually raised to 240 °C. Helium was used as the carrier gas at a flow of 1 mL min−1. The identification of each fatty acid was carried out by comparing the retention time with that of the corresponding standard methyl ester (Sigma-Aldrich, St. Louis, MO, USA). The amount of single fatty acids was expressed as area % with respect to the total area [23 ,31 (link)].
3
Fatty Acid Profiling by Gas Chromatography
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Fatty acid analysis was done on a Perkin Elmer gas chromatograph 8700 fitted with FID detector. SP-2340 fused silica capillary column (30 m × 0.25 mm × 0.2 µm) (Supelco, Inc., Bellefonte, PA) was employed, with temperature programmed from 120˚C to 220˚C at 4˚C/min. Nitrogen as carrier gas was used at a flow rate of 3.5 mL/min. The injector and detector temperature were set at 260˚C and 270˚C respectively. Fatty acids were identified by comparison of their retention times with that of the authentic standard and retention times. A built-in data handling program provided by the manufacturer of the gas chromatograph (Perkin-Elmer) was used for all quantifications as reported earlier [16] (link).
4
Analyzing Fungal Fatty Acid Profiles
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Total lipids were extracted from whole-cell homogenates and methylated by following previously described approaches (He et al., 2018b (link)). Briefly, A. oryzae mycelia were collected, washed with distilled water, and freeze dried. After that, the mycelia of each group were powdered and weighed. The same weight of mycelia powder was subjected to lipid extraction. The lipid extracts were incubated in chloroform with 2% H2SO4–MeOH solution at 70°C for 2 h to obtain fatty acid methyl esters (FAMEs). FAME components were separated and analyzed using coupled QP2010 gas chromatography-mass spectrometry (GC-MS) (Shimadzu, Kyoto, Japan). The system was equipped with Supelco SP-2340 fused silica capillary column (30 m × 0.25 mm i.d., with film thickness of 0.2 μm; Bellefonte, PA, United States). FAMEs were identified by comparing their mass spectra with a spectrum database. FA peaks were identified based on comparisons of their retention times to the external standards or similarity search. The relative amounts of individual FA components were performed by using the peak area of the most intensive ion of each peak, and the percentage of UFAs was also calculated (Kim and Salzberg, 2011 (link); He et al., 2018b (link)).
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