Total lipids of the diets (5 g) and i.m. adipose tissue (100 mg) were extracted and fatty acid methyl esters (FAME) were prepared as described previously (Archibeque et al., 2005 (link)). The diets were sampled at three intervals and the data pooled (Table 2 ). FAME were analyzed by GC-FID (model CP-3800 equipped with a CP-8200 auto-sampler; Varian Inc, Palo Alto, CA, USA). Separation of FAME was accomplished on a fused silica capillary column (100 m×0.25 mm ID) (model CP-3800, Varian Inc, USA) with the helium as carrier gas (flow rate = 1.7 mL/min). One microliter of sample was injected with the split ratio of 100:1 at 270°C. Oven temperature was set at 165°C for 65 min and then increased to 235°C (°C/min) and held for 15 min. Flame ionization detector (FID) detected the signal at 270°C. An authentic standard (GLC 68D, Nu-chek Prep, Waterville, MN, USA) was used to confirm the identity of each peak.
Model cp 3800
Manufactured by Agilent Technologies
Sourced in United States
The Agilent CP-3800 is a gas chromatograph designed for high-performance separation and analysis of complex mixtures. It features advanced hardware and software capabilities to deliver reliable and accurate results.
Automatically generated - may contain errors
Lab products found in correlation
Cp 8200 auto sampler, by Agilent Technologies (1 mentions)
Glc 68d, by Nu-Chek Prep (1 mentions)
Cary 50 bio uv visible spectrophotometer, by Agilent Technologies (1 mentions)
Cary 50 mpr microplate reader, by Agilent Technologies (1 mentions)
Uv 1600pc, by Avantor (1 mentions)
Saturn 2200, by Agilent Technologies (1 mentions)
Vf 5m, by Agilent Technologies (1 mentions)
5 protocols using model cp 3800
1
Determination of Dietary Fatty Acids
2
Essential Oil Chemical Profiling by GC-MS
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The EO was diluted with n-hexane (GC grade, 5 μL:1 mL) and 5 μL were injected into the GC (GC, Model CP-3800, Varian, Walnut Creek, CA, USA) and linked with a mass spectrometer (MS, Model Saturn 2200, Varian, Walnut Creek, CA, USA) equipped with a VF-5ms-fused silica capillary column (5% phenyl-dimethylpolysiloxane, with dimensions of 30 m × 0.25, film thickness was 0.25 μm, Varian). An electron impact (EI) ionization detector was used, with an ionization energy of 70 eV. Carrier gas (helium) was adjusted to have a steady flow rate (1 mL/min). The temperature of the oven was programed as follows: 1, 50, and 5 min at 50, 230, and 290 °C, respectively. The split ratio of injection samples was 1/500, with total time equal to 60 minutes. Identification of the constituents was conducted by comparison of Kovat’s retention indices (RI) relative to a set of co-injected standard hydrocarbons (C10–C28, Sigma-Aldrich, Darmstadt, Germany) [52 (link)]. Components were identified by comparing their MS data and their corresponding retention indices with the Wiley Registry of Mass Spectral Data 10th edition (April 2013), the NIST 11 Mass Spectral Library (NIST11/2011/EPA/NIH), and literature data [53 ]. All of the identified constituents and their relative abundance percentages are listed in Table 1 .
3
Nutrient Composition Analysis in Animal Digesta
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All digesta and excreta samples were freeze-dried and finely ground using a centrifuge mill with a 0.5 mm screen (Model ZM 200, Retsch, Hann, Germany). Samples were analysed for dry matter and NSP, uronic acids and TiO2. Dry matter was determined according to the standard method of AOAC (2012) (Method 930.15). The constituent sugar components of NSP were determined as alditol acetates using gas–liquid chromatography (Model CP3800, Varian Inc., Palo Alto, CA), according to the method described by Englyst et al. (1994) (link) with some modifications (Theander et al., 1995 (link); Morgan et al., 2019 (link)). Uronic acids were determined by spectrophotometry, at 400 and 450 nm (UV-1600PC, VWR, Darmstadt, Germany), using the method illustrated by Scott (1979) . Quantification of TiO2 was conducted using UV-spectroscopy at 410 nm (Cary 50 Bio UV–Visible spectrophotometer equipped with a Cary 50 MPR microplate reader, Varian Inc., Palo Alto, CA), using the method of Short et al. (1996) . All the analysed values were corrected for dry matter percentages.
4
GC-MS Analysis of LGBME Compounds
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LGBME was analyzed by GC–MS analysis technique performed by GC (Model CP-3800, Varian, Santa Clara, CA, USA) with MS (model: Varian Saturn-2200) spectrometer equipped with a flame-ionization detector and capillary column with VF-5 ms (30 m × 0.25 mm, 0.25 μm). The instrument was operated using electron-impact-ionization mode under specified conditions (ionization voltage −70 eV, detector temperature 280 °C, and injector temperature 250 °C). Helium, an inert gas, was used as a carrier gas, the flow rate was 1 mL/minute, and the sample injection volume was 1 μL. The initial temperature for the column was 40 °C (1 min), and the final temperature was 310 °C at a gradually increasing rate of 10 °C/minute. Temperature was held constant at 310 °C for 10 min. Chemical compounds were identified by their GC retention time, retention indices relative to n-alkanes, and comparison of their mass spectra with the NIST 2008 (National Institute Standard and Technology 2008) Library data.
5
GC-MS Analysis of Cell Suspension Extracts
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Hydro distillate extract from a cell suspension after 40 days was diluted with n-hexane (GC grade, 2 μL:1 mL) and injected (1 μL) using an auto-sampler injector (Model Combi Pal, Varian) to the GC–MS (GC, Model CP-3800, Varian, Walnut Creek, CA, USA) linked with a mass spectrometer (MS, Model Saturn 2200, Varian) using a VF-5ms fused silica capillary column (5% phenyl- dimethylpolysiloxane, 30 m × 0.25 mm i.d., film thickness 0.25 μm, Varian, Palo Alto, CA, USA). The electron impact (EI) ionization detector was used with an ionization energy of 70 eV. Helium was a carrier gas (99.99%) with a constant rate (1 mL/min). The injector and mass transfer line temperatures were 240 and 300 °C, respectively. The oven temperature was held at 50 °C for 1 min, raised to 230 °C for 50 min at 30 °C/min, finally raised to 290 °C for 5 min at 10 °C/min and held isothermal for 6 min. The sample split injection ratio was 1/500, with a total time of 54.3 min. The identification of components was based on matching with a mixed standard (n-alkanes (C6–C26)) and the Wiley and National Institute of Standards and Technology (NIST) electronic library.
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