Fatty acid methyl esters (FAMEs) from salmon lecithin were prepared as described by Ackman (1998) [24 (link)]. Subsequently, FAMEs were analyzed while using a Shimadzu 2010 gas chromatography (Shimadzu, Marne-la-Vallée, France) system that was equipped with a flame-ionization detector. FAME was injected into a fused silica capillary column (60 m, 0.25 mm i.d. × 0.20 µm film thicknesses, SPTM2380 Supelco, Bellfonte, PA, USA). Injector and detector temperatures were settled at 250 °C. The column temperature was fixed initially at 120 °C for 3 min., then raised to 180 °C at a rate of 2 °C min.−1, and maintained at 220 °C for 25 min. Individual fatty acids were identified while using standard mixtures (PUFA1 from a marine source and PUFA2 from a vegetable source; Supelco, Sigma-Aldrich, Bellefonte, PA, USA). The results are shown as mean ± SD of triplicate analyses.
Pufa 1
Manufactured by Merck Group
Sourced in United States
PUFA-1 is a laboratory instrument designed for the analysis of polyunsaturated fatty acids (PUFAs). It employs advanced chromatographic techniques to separate and quantify individual PUFA species within a sample. The core function of PUFA-1 is to provide accurate and reliable data on the PUFA composition of various biological and food samples.
Automatically generated - may contain errors
Lab products found in correlation
Pufa 3, by Merck Group (6 mentions)
Menhaden oil, by Merck Group (3 mentions)
Pufa2, by Merck Group (3 mentions)
Star 3800 cp gas chromatograph, by Agilent Technologies (2 mentions)
Supelco 37 component fame mix, by Merck Group (2 mentions)
Sptm 2380, by Merck Group (1 mentions)
Gc 17a, by Shimadzu (1 mentions)
Omegawax 320, by Merck Group (1 mentions)
Db wax, by Agilent Technologies (1 mentions)
Hp 6890 chromatograph, by Agilent Technologies (1 mentions)
60 m db 23 capillary column, by Agilent Technologies (1 mentions)
Agilent autosampler, by Agilent Technologies (1 mentions)
Trans vaccenic acid, by Merck Group (1 mentions)
Linoleic acid, by Merck Group (1 mentions)
11 protocols using pufa 1
1
Quantifying Salmon Lecithin Fatty Acids
2
Fatty Acid Profiling of Seafood Lipids
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The lipid to be used for fatty acid profile determination was extracted from 100 g of ground samples according to Bligh & Dyer (1959) . The extracted lipid was stored under nitrogen in the dark at À20 C for further analyses. The coating oil medium of salted-ripened fillets, canned and marinated anchovy was also analysed. Fatty acid profile was determined by fatty acid methyl ester (FAMEs)/gas chromatography (GC). For this, 100 mg of the lipid sample was dissolved in hexane and treated with a solution of KOH in methanol according to the norm ISO 5509 (2000) . Fatty acids methyl esters were separated and quantified using a gas chromatograph (Shimadzu GC-17A, Japan) equipped with a 30-m fused silica capillary column Omegawax 320 (Supelco Inc., Bellefonte, PA) (0.32 mm i.d.; 0.25 mm film) and a flame ionization detector. Carrier gas was nitrogen. Sample injected volume was 1 ml. Column oven temperature was programmed to begin at 150 C, increase to 225 C at 1.5 C/min and held for 13 min. The temperature of the injector port and the detector was held at 250 C. Peak retention times and area percentages of total fatty acids were identified by comparison with the standard PUFA-1, Marine Source from Supelco (Bellefonte, PA). The polyene index (PI) was calculated as the following fatty acid ratio: (C20:5o3 + C22:6o3)/C16:0 (Lubis & Buckle, 1990) .
3
Fatty Acid Profiling of Bioaccessible Fraction
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The fatty acid profile was determined in the samples before and after digestion (bioaccessible fraction). Fatty acid methyl esters (FAME) were prepared by acid‐catalyzed transesterification using the methodology described by Bandarra, Batista, Nunes, Empis, and Christie (1997 ). Samples were injected into a Varian Star 3800 Cp gas chromatograph (Walnut Creek, CA, USA), equipped with an autosampler with a flame ionization detector at 250°C. FAME were identified by comparing their retention time with those of Sigma‐Aldrich standards (PUFA‐3, Menhaden oil, and PUFA‐1, Marine source from Supelco Analytical). Data in mg/100 g of edible part were calculated using the peak area ratio (% of total fatty acids) and the lipid conversion factors set by a previous study (Weihrauch, Posati, Anderson, & Exler, 1977 ).
4
Fatty Acid Profiling and Cardiovascular Risk
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Fatty acid methyl esters (FAMEs) were prepared from the freeze-dried ingredients, experimental products, and bioaccessible fractions by acid-catalyzed transesterification [11 (link)]. The samples were applied to a DB-WAX (Agilent, Santa Clara, CA, USA) capillary column (film thickness: 0.25 μm, 30 m × 0.25 mm i.d.) integrated into a Varian Star 3800 CP gas chromatograph (Walnut Creek, CA, USA). The FAMEs were identified by comparing their retention times with those of several Sigma-Aldrich standards (PUFA-3, Menhaden oil, and PUFA-1, a marine source standard from Supelco, Bellefonte, PA, USA). The LOD was 1 mg/100 g. Data in mg/100 g of edible parts were calculated using the peak area ratio (% of total FA) and the lipid conversion factors [17 (link)]. Analyses were always performed in triplicate.
For an assessment of the dietary impact of the FA composition on the incidence of coronary heart disease, the index of atherogenicity (IA) and the index of thrombogenicity (IT) were calculated as proposed by Ulbricht and Southgate (1991) [18 (link)]:
where MUFA is the monounsaturated FA.
For an assessment of the dietary impact of the FA composition on the incidence of coronary heart disease, the index of atherogenicity (IA) and the index of thrombogenicity (IT) were calculated as proposed by Ulbricht and Southgate (1991) [18 (link)]:
where MUFA is the monounsaturated FA.
5
Fatty Acid Methyl Esters (FAME) Quantification
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Fatty acid methyl esters (FAME’s) were prepared by acid-catalyzed transesterification using the methodology described by Bandarra et al. [21 (link)]. Samples were injected into a Varian Star 3800 CP gas chromatograph (Walnut Creek, CA, USA and equipped with an auto sampler with a flame ionization detector at 250 °C. FAME’s were identified by comparing their retention times with those of Sigma–Aldrich standards (PUFA-3, Menhaden oil, and PUFA-1, Marine source from Supelco Analytical).
6
Chromatographic Profiling of Fatty Acids
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Fatty acid methyl esters (FAMEs) were prepared as described by Ackman [20] . The separation of FAMEs was carried out on a Shimadzu 2010 gas chromatography (Perichrom, Saulx-lès-Chartreux, France) equipped with a flame-ionization detector. A fused silica capillary column was used (60 m, 0.2 mm i.d.×0.25 µm film thicknesses, SP™2380 Sopulco, Bellfonte-PA-USA). Injector and detector temperatures were set at 250°C. The column temperature was set initially at 120°C for 3 min, then rose to 180°C at a rate of 2°C/min and held at 220°C for 25 min. Standard mixtures (PUFA1 from marine source and PUFA2 from vegetable source; Supelco, Sigma–Aldrich, Bellefonte, PA, USA) were used to identify fatty acids. The results were presented as triplicate analyses.
7
Fatty Acid Methyl Esters Analysis
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Fatty acid methyl esters (FAMEs) were analyzed as previously described, according to the amount of lipids [71 (link),72 (link)]. FAME separation was carried out via gas chromatography (Perichrom, Saulx-lès-Chartreux, France). Injector and detector temperatures were fixed at 250 °C. Column temperature was initially set at 120 °C for 3 min, then it was increased to 180 °C at a rate of 2 °C min−1 and kept at 220 °C for 25 min. Standard mixtures (PUFA1 from marine source and PUFA2 from vegetable source, Supelco, Sigma–Aldrich, Bellefonte, PA, USA) and a nonadecanoic acid internal control were used to categorize fatty acids. All runs were performed in triplicate.
8
Fatty Acid Profiling of Microalgae
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Fatty acid methyl esters (FAME's) were prepared from the freeze-dried microalgae, feeds, and the experimental organism (rotifers and artemia) biomass by acid-catalysed transesterification using the methodology described by Bandarra et al. (1997) . The FAMEs were identified by comparing their retention time with those of several Sigma-Aldrich standards (PUFA-3, Menhaden oil, and PUFA-1, marine source from Supelco Analytical). The LOD is 1 mg/100g. Results were calculated in % of total fatty acids on the basis of peak areas and results in mg/g were attained through the internal standard (10 mg/ml of heneicosanoic acid, 21:0) method. Analyses were always done in triplicate.
9
Fatty Acid Composition Analysis of Oils
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To evaluate fatty acid composition of oils obtained from A. vulgare, C. nucifera, and E. guineensis, alkaline trans-methylation of fatty acids was carried out as described by Raes et al.100 (link). Analysis of methyl esters was performed using gas chromatography (GC) with the HP 6890 chromatograph (Agilent Technologies, Inc., Santa Clara, CA, USA), equipped with a 60 m DB-23 capillary column (60 m × 0.25 mm × 0.25 μm) and a flame-ionization detector (FID); split injections were performed using an Agilent autosampler. A total of 1 µL of standards in hexane were injected in split mode (1:40 ratio) into the injector, heated to 230 °C. The column temperature was initially set at 120 °C for 6 min then programmed to 170 °C at a rate of 15 °C/min. The temperate gradient was further configured to 210 °C at the rate of 3 °C/min and held for 13.5 min. Finally, the temperature was programmed to 230 °C at the rate of 40 °C/min and held for 7 min. Nitrogen was used as the carrier gas, at a flow rate of 0.8 mL/min. Supelco 37 Component FAME Mix, PUFA 1, PUFA 2, PUFA 3, trans-vaccenic acid, and a mixture of conjugated isomers of linoleic acid (Sigma-Aldrich, Prague, CZ) were used as standards. Fatty acids were identified based on retention times with respect to standards.
10
Fatty Acid Composition Analysis
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Fatty acid composition was determined by methanol extraction on 300 mg wet biomass. Pellet suspension in methanol (5 mL/g pellet) was sonicated (5 min at 37 kHz) prior to centrifugation at 3000× g for 6 min at 4 °C. The supernatant was recovered and the residue was extracted another two times using the same procedure. Supernatants were combined and dried. Part of the oily residue (0.5 mg) was saponified by catalytic amount of Na2CO3 in 500 μL MeOH at room temperature overnight under magnetic stirring. After neutralization by 1% acetic acid, the methyl esters were extracted by partition between diethyl ether and water. The ether layer was dried under nitrogen and the resulting material was dissolved in n-hexane (1 μg μL−1) and analyzed by GC-MS (70 eV) on a 5% diphenyl column (inlet temperature of 270 °C, transfer line set at 280 °C and ion source temperature of 250 °C). Elution of fatty acid methyl esters (FAMEs) was accomplished by an increasing gradient of temperature according to the following program: 200 °C for 2.5 min then 15 °C/min up to 290 °C, followed by 7 min at 290 °C. Each sample (2 μL) was directly injected. Fatty acids were identified by comparing retention time (RT) of peaks to standard fatty acid methyl asters (PUFA-1, Marine Source, Sigma-Aldrich).
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