Intramuscular lipids were extracted from the fillet muscles in duplicates using the method of Folch et al. [33 (link)] with modifications by Shin and Ajuwon [34 (link)]. Briefly, fatty acid methyl esters (FAME) were prepared by a trans-esterification reaction, after which FAMEs were extracted in hexane. FAMEs were analyzed on a gas chromatograph (Varian CP 3900 with CP-8400 autosampler, Agilent, Santa Clara, CA, USA) equipped with a 105 m Rtx-2330 fused silica capillary GC column (10729, Restek, Bellefonte, PA, USA). Fatty acids were identified by comparison of retention times with the known standards (Supelco 37 components FAME Mix, Sigma Aldrich, St. Louis, MO, USA). Detected fatty acids were expressed in grams per 100 grams of intramuscular lipid.
Supelco 37 components fame mix
Manufactured by Merck Group
Sourced in United States
Supelco 37 components FAME Mix is a laboratory standard mixture containing 37 fatty acid methyl esters (FAMEs) at known concentrations. It is used for the identification and quantification of fatty acids in various samples through gas chromatography analysis.
Automatically generated - may contain errors
Lab products found in correlation
Sp 2560, by Merck Group (2 mentions)
Agilent 6890n gc, by Merck Group (2 mentions)
Triheptadecanoin, by Merck Group (1 mentions)
Clarus 500, by PerkinElmer (1 mentions)
Hp6890n, by Hewlett-Packard (1 mentions)
Hp7683, by Hewlett-Packard (1 mentions)
Rtx 2330, by Restek (1 mentions)
Pheophorbide a, by Cayman Chemical (1 mentions)
Avance dpx 400 spectrometer, by Bruker (1 mentions)
Lutein, by Cayman Chemical (1 mentions)
Tlc silica gel 60 f254 plate, by Merck Group (1 mentions)
Anhydrous sodium sulfate, by Junsei (1 mentions)
Potassium hydroxide, by Daejung Chemicals (1 mentions)
Triundecanoin, by Nu-Chek Prep (1 mentions)
10 protocols using supelco 37 components fame mix
1
Intramuscular Lipid Extraction and GC Analysis
2
Quantifying Fatty Acid Composition in White Adipose Tissue
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Total lipids from WAT samples were extracted with chloroform-methanol 2:1 (v/v) according to Folch et al. [59 (link)]. FA methyl esters were prepared with 2% methanolic HCl at 100°C for 2 hours, and extracted with hexane after addition of 2% sodium bicarbonate. All reagents were added with butylated hydroxy toluene (BHT; 25 mg/L) to avoid autoxidation of PUFA [60 (link)]. FA methyl esters were analyzed using a Perkin Elmer Clarus 500 gas chromatograph, as previously described [18 (link)]. Peaks were identified by comparison of their retention times with FA methyl ester standards (Supelco 37 Components FAME Mix, Sigma-Aldrich) and quantified respect to triheptadecanoin (Sigma-Aldrich) used as internal standard (IS). The individual FA detected were expressed as a percent of total FA.
3
Intramuscular Lipid Extraction and Fatty Acid Analysis
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Intramuscular lipids were extracted in duplicate from powdered samples using the method described by Folch et al. [34 (link)] with modifications described by Shin and Ajuwon [35 (link)]. Only two birds from each pen that were closest to pen average live body weight were chosen to sample for this portion. Fatty acid methyl esters (FAME) were prepared from the extracted lipids by adding sodium methoxide to methanol. The FAME were analyzed using a gas chromatograph (Varian CP 3900) equipped with a 105 m Rtx-2330 (Restek) fused silica capillary CG column (0.22 mm ID and 0.20 µm df). Helium was used as the carrier gas with a flow rate of 40 mL/min. Injector and detector temperatures were at 260 °C. Injection volume was set at 1 µL with a 50:1 split injection. The column oven temperatures were increased from 140 °C to 180 °C at a rate of 8 °C/min, from 180 °C to 260 °C at a rate of 5 °C/min, and then held at 260 °C for 15 min. The fatty acids were identified by comparing them to a retention time of a known standard (Supelco 37 components FAME Mix, Sigma-Aldrich, St. Louis, MO, USA) and the peak area of the fatty acid detected was expressed as a percent of the total peak area.
4
Lipid Extraction and Fatty Acid Analysis
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The extraction of lipid from muscle samples was carried out according to method
described by Folch et al. (1957) (link). The
lipid methyl esters were evaluated by treating with 1.0 N methanolic NaOH and
methylated by boron trifluoride in methanol solution. Fatty acid methyl ester
(FAME) determination was conducted by gas chromatography HP6890N
(Hewlett-Packard, Santa Clara, CA, USA) using an automatic sampler HP7683
(Hewlett-Packard). The separation of FAME from the samples was conducted under
the following conditions: Column oven temperature was increased from 50°C
to 180°C at 10°C per min and maintained at 180°C for 20
min, the temperature of both the injector and detector was 250°C, and the
sample injection volume was 1 μL. Finally, resulted each fatty acids
content was recognized by comparing the retention times to those of standards
FAME mixture (Supelco 37 Components FAME Mix, Sigma-Aldrich, St. Louis, MO,
USA). The results are expressed as the percentage of total fatty acids detected
based on the total peak area.
described by Folch et al. (1957) (link). The
lipid methyl esters were evaluated by treating with 1.0 N methanolic NaOH and
methylated by boron trifluoride in methanol solution. Fatty acid methyl ester
(FAME) determination was conducted by gas chromatography HP6890N
(Hewlett-Packard, Santa Clara, CA, USA) using an automatic sampler HP7683
(Hewlett-Packard). The separation of FAME from the samples was conducted under
the following conditions: Column oven temperature was increased from 50°C
to 180°C at 10°C per min and maintained at 180°C for 20
min, the temperature of both the injector and detector was 250°C, and the
sample injection volume was 1 μL. Finally, resulted each fatty acids
content was recognized by comparing the retention times to those of standards
FAME mixture (Supelco 37 Components FAME Mix, Sigma-Aldrich, St. Louis, MO,
USA). The results are expressed as the percentage of total fatty acids detected
based on the total peak area.
5
Intramuscular Lipid Extraction Protocol
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Extraction of intramuscular lipids was performed in duplicate from loin samples at 1 d postmortem in accordance to Folch, Lees, and Sloane Stanley [33 (link)]. In short, 1.0 g of pulverized muscle tissue was homogenized with 21 mL of 2:1 (v/v) chloroform to methanol. Sodium methoxide in methanol (0.5 N) was added to prepare fatty acid methyl esters (FAME). Fatty acid profiling was conducted using the conditions described by Tuell et al. [32 (link)]. Identification of fatty acids was conducted by comparing retention time to known standards (Supelco 37 components FAME mix, Sigma Aldrich, St. Louis, MO, USA). Fatty acids were expressed as g of fatty acid per 100 g of intramuscular lipid.
6
Fatty Acid Methyl Esters Analysis by GC
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The preparation of fatty acid methyl esters (FAME ) concentration was performed using sodium methoxide in methanol (Hartman and Lago, 1973 (link)). The FAME was then reconstituted in chromatography grade hexane for gas chromatography (Varian CP 3900, Varian Analytical Instruments, Walnut Greek, CA, UAS). The gas chromatograph (GC) was equipped with a 105 m Rtx-2330 (Restek, Bellefonte, PA, USA) fused silica capillary GC column (0.22 mm I.D. and 0.20 μm of film thickness). Helium was used as the carrier gas at a flow rate of 40 mL/min. Injector and detector temperatures were 260°C. The injected volume was 1 μL at a 50:1 split injection. Column oven temperature was increased from 140 to 180°C at a rate of 8°C/min, from 180 to 260°C at a rate of 5°C/min and then held at 260°C for 15 min. Fatty acids were identified by comparison of retention time of a known standard (Supelco 37 components FAME Mix, Sigma-Aldrich, USA) and expressed as a ratio: the peak area of the detected fatty acid to the total peak area x 100%. The results of saturated fatty acids, UFA, mono- and poly-unsaturated fatty acids (MUFA and PUFA ) were calculated based on the content of the whole set of FA profile (%).
7
Phytochemical Analysis of Pigments
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The NMR spectra using standard pulse programs were recorded at room temperature on a Bruker Avance DPX-400 spectrometer (Bruker, Billerica, MA, USA) operating at 400 (1H) and 100 (13C) MHz. The chemical shift (δ, ppm) values were calibrated using the residual NMR solvent and coupling constant (J) was reported in Hertz (Hz). Column chromatography was done on normal-phase silica gel (230 × 400 mesh, J. T. Baker, Center Valley, PA, USA) or reversed-phase silica gel (C18, 40 μm, J. T. Baker). Silica gel 60 F254 TLC plates (Merck, Darmstadt, Germany) and reversed-phase TLC plates (C18, Merck, Darmstadt, Germany) were used for analytical TLC. The plates were visualized by spraying 10% H2SO4 followed by heating. The authentic samples pheophorbide a (>95%) and lutein (>90%) were purchased from Cayman Chemical Company (Ann Arbor, MI, USA) and Acros Organics (Pittsburgh, PA, USA), respectively. Their structure and purity were further confirmed by NMR and HPLC in our laboratory. The 37 standards of fatty acid methyl esters (Supelco® 37 components FAME mix) used for GC-MS analysis were purchased from Sigma-Aldrich (St. Louis, MO, USA).
8
Herring Oil Fatty Acid Composition Analysis
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The chemical composition of air-dried herring oil was determined by GC using an Agilent 6890N GC and SP-2560 (Supelco, Sigma-Aldrich, St. Louis, MO, United States) fused silica capillary column (100 m × 0.25 mm i.d., film thickness 0.25 μm). Capillary column details, temperature conditions, and the derivatization (methylation) of fatty acids were as previously described (Lee et al., 2017 (link)). In brief, helium was used as the carrier at 0.75 ml/min and the GC injector was held at 225°C. The GC oven temperature was programmed as follows; 100°C for 4 min, increased to 240°C at 3°C/min, and followed by 15 min at 250°C. The split ratio was controlled at 200:1. Triundecanoin (C11:00) was used as the internal standard and quantifications were performed by integrating areas and correcting for fatty acid methylation. Supelco 37 components FAME Mix (Supelco) were used as the reference standard.
9
Extraction and Analysis of Fatty Acids and Carotenoids
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Potassium hydroxide, sodium chloride, hexane, ethyl acetate, chloroform, hydrogen chloride, diethyl ether, petroleum ether, and butylated hydroxytoluene (BHT) were purchased from Daejung Chemicals & Metals Co., Ltd. (Shiheung, Korea). Anhydrous sodium sulfate was purchased from Junsei Chemical Co., Ltd. (Tokyo, Japan). Triundecanoin (C11:0, internal standard for FA analysis) was obtained from Nu-Chek Prep, Inc. (Elysian, MN, USA). The solvents (i.e., acetonitrile, methylene chloride, hexane, methyl alcohol, iso-octane, and chloroform) used for high-performance liquid chromatography (HPLC) and gas chromatography (GC) analysis were acquired from Fisher Scientific Korea (Seoul, Korea). BF3-methanol (14%, w/w), pyrogallol, and analytical standards of FA methyl esters (Supelco 37 components FAME mix), lutein (07168-1MG), zeaxanthin (14681-1MG-F), β-cryptoxanthin (C6368-1MG), lycopene (75051-10MG), α-carotene (50887-1MG), and β-carotene (C4582-10MG) were purchased from Sigma-Aldrich Korea (Seoul, Korea).
10
GC-MS Analysis of Saw Palmetto Oil
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The components of saw palmetto oil were separated and analysed by gas chromatography‐mass spectroscopy (GC‐MS) using an Agilent 6890N GC and SP‐2560 (Supelco, Sigma‐Aldrich, St. Louis, USA) with a silica capillary column (100 m × 0.25 mm i.d., film thickness 0.25 mm). Column temperature conditions and the derivatization (methylation) of fatty acids were as previously described (Kim et al., 2018 (link)). Triundecanoin (C11:0) was used as the internal standard, and quantifications were performed by integrating areas and correcting for fatty acid methylation. Supelco 37 components FAME Mix (Supelco) were used as the reference standard.
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