The total fat content (liver and muscle) and fatty acid profile (added dietary fats, liver, and muscle) analyses were also performed at the ARC, Irene Analytical Services Laboratories, South Africa. Tissue samples from all birds in a specific dietary group were pooled, freeze dried and then milled and analysed as composite, representative sample from each group. Total fat content was determined by the Soxhlet method as described by the Official Methods of Analysis of Analytical Chemists (2005; method number 920.39) [12 ]. For the fatty acid profile analyses, gas chromatography was used (HP6890 GC, Hewlett Packard, Bristol, UK) and nonadecanoic acid (C19:0) was used as the internal standard. Only the fatty acids that accounted for >1% of the total fatty acids in the respective tissue lipids are reported in the results tables.
Hp6890 gc
Manufactured by Hewlett-Packard
Sourced in United Kingdom, United States
The HP6890 GC is a gas chromatograph designed for the separation, identification, and quantification of chemical compounds. It features a programmable oven, a variety of detectors, and advanced control and data analysis software. The HP6890 GC is a versatile instrument suitable for a wide range of applications in analytical chemistry and related fields.
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Lab products found in correlation
7890a gc system, by Agilent Technologies (1 mentions)
Hp5973 mass spectrometer, by Hewlett-Packard (1 mentions)
Db 5 capillary column, by Agilent Technologies (1 mentions)
Db waxetr, by Agilent Technologies (1 mentions)
Db 23 capillary column, by Merck Group (1 mentions)
Chemstation software, by Agilent Technologies (1 mentions)
Hp 5ms, by Hewlett-Packard (1 mentions)
9 protocols using hp6890 gc
1
Analyzing Fat Content and Fatty Acids
2
GC-MS Analysis of Unknown Compounds
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GC-MS analysis was performed on an HP6890 GC coupled with a HP5973 mass spectrometer (Hewlett Packard, CA, USA). The column was a HP-5MS fused-silica capillary column (50 m x 0.25 mm i.d.; 0.25-μm film thickness) with helium as the carrier gas, and it was run at a constant pressure of 9.78 psi. Injection was conducted using a splitless mode at an injector temperature of 250 °C. The oven temperature was ramped from 40 °C to 280 °C (1-min hold) at a rate of 25 °C/min. The oven temperature was held at 310 °C for 6 min for each analysis. The total run time for each sample was approximately 28 min. The GC-MS interface temperature was set to 280 °C. MS mode was used during analytical scanning from 20 to 650 atomic mass units (amu). The ion source temperature was set at 250 °C.
The blank was injected first each time, followed by the sample injection. The chromatograms obtained from the total ion current (TIC) were integrated without any correction for co-eluting peaks, and the results were expressed as total abundance. The TIC peaks and chromatograms were analyzed using an Agilent Technologies 7890A GC system with 597C VL MSD. All peaks were identified based on mass spectral matching (≥ 90 %) from both the National Institute of Standards and Technology (NIST) and Wiley libraries. Only compounds with 90 % or greater spectral matching accuracy were reported.
The blank was injected first each time, followed by the sample injection. The chromatograms obtained from the total ion current (TIC) were integrated without any correction for co-eluting peaks, and the results were expressed as total abundance. The TIC peaks and chromatograms were analyzed using an Agilent Technologies 7890A GC system with 597C VL MSD. All peaks were identified based on mass spectral matching (≥ 90 %) from both the National Institute of Standards and Technology (NIST) and Wiley libraries. Only compounds with 90 % or greater spectral matching accuracy were reported.
3
GC-FID and GC-MS Analysis of Headspace Volatiles
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The GC and electronic impact (EI) GC–MS systems used were as described (Zhang et al. 2004 (link)). A Hewlett Packard (HP) 6890 GC was coupled to a flame ionization detector (FID) using DB-WAXETR or DB-5 capillary column (60 m × 0.25 mm i.d., 0.25-μm film thickness, J&W Scientific Inc., Folsom, CA). Oven temperature was started at 50 °C for 2 min, then programmed to rise to 230 °C at 15 °C/min, and held for 20 min in the splitless mode with hydrogen as carrier (2 mL/min). For GC–MS analysis, a HP 6890 GC was coupled to a HP 5973 Mass Selective Detector (MSD) using the same columns as GC-FID, but with helium as carrier (1.4 mL/min). A 70 eV electron beam was employed for sample ionization. The chemical identification of the headspace volatiles was based on comparison of their mass spectra with the NIST and Wiley mass spectral libraries, and identities were confirmed by mass spectra and GC retention times of authentic standards on both polar and nonpolar GC capillary columns (Zhang et al. 1999 (link)). The ratios of identified components were determined by using GC–FID.
4
GC-MS Analysis of Cuticular Hydrocarbons
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GC-MS analysis of extracted CHCs was carried out on a Hewlett-Packard (HP) 6890 GC (Hewlett-Packard, Sunnyvale, CA) in splitless mode, interfaced to an HP 5973 mass selective detector (MSD), with helium carrier gas. The column was programmed from 100°C/2 min, 50°C/min to 250°C, then 250 to 320°C at 4°C/min. Injector and transfer line temperatures were 320°C. Prior to GC-MS analysis, samples were removed from the refrigerator, concentrated to dryness under a steady stream of nitrogen, and resuspended in 30 μl of hexane with internal standard from which one μl was injected into GC-MS for analysis. A control sample of hexane was run through the GC-MS every day before samples were analyzed to confirm that the GC column was clean. The adults and egg clusters were analyzed as individual replicates from each strain. Hydrocarbon peaks were identified based on their relative retention time. The abundance of each identified hydrocarbon peak was calculated relative to the internal standard.
5
Asparagus Cell Wall Composition Analysis
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Asparagus spear sections were separately ground with a mortar and pestle in liquid nitrogen and washed successively with 80% ethanol, acetone and methanol to generate alcohol insoluble residues (AIR) as described by Pettolino et al. [38 (link)]. The neutral sugar (alditol acetates) and polysaccharide (linkage analysis) compositions of the AIR were determined by GC-MS (Hewlett-Packard HP 6890 GC with a Hewlett Packard 5973 MS, Agilent) following the procedure described by Pettolino et al. [38 (link)].
Lignin content was determined gravimetrically as described by Luo et al. [39 ]. The final lignin-containing residue was weighed and the Klason-lignin content recorded as grams per 100 g FW.
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Asparagus spear sections were separately ground with a mortar and pestle in liquid nitrogen and washed successively with 80% ethanol, acetone and methanol to generate alcohol insoluble residues (AIR) as described by Pettolino et al. [38 (link)]. The neutral sugar (alditol acetates) and polysaccharide (linkage analysis) compositions of the AIR were determined by GC-MS (Hewlett-Packard HP 6890 GC with a Hewlett Packard 5973 MS, Agilent) following the procedure described by Pettolino et al. [38 (link)].
Lignin content was determined gravimetrically as described by Luo et al. [39 ]. The final lignin-containing residue was weighed and the Klason-lignin content recorded as grams per 100 g FW.
6
Fatty Acid Profiling of Meat Extracts
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The Soxhlet Apparatus was used to extract the oil from the breast muscle sample as described by the Association of Analytical Chemists (AOAC, 2006 ; method number: 920.39). The fatty acid profiles of the oil extracted from the meat were determined as described by Christopherson & Glass, (1969) . Briefly, the oil extracts were trans-methylated with 2 mol/L methanol sodium hydroxide. The resulting fatty acid methyl esters were extracted in heptane, filtered (Nylon syringe filters, pore size: 0.45 µm, diameter: 13 mm with a glass fibre prefilter, Membrane Solutions) and dried under nitrogen after which they were separated by a temperature gradient over 45 min on a gas chromatograph with nitrogen as carrier gas on a DB-23 capillary column (90 cm × 250 μm × 0.25 μm; Supelco, Sigma-Aldrich). The gas chromatograph consisted of an HP6890 GC (Hewlett Packard, Bristol, UK) with a flame ionisation detector. Both the detector and injector temperatures were set at 300 °C. A personal computer equipped with Chemstation software (Agilent Technologies Inc., Santa Clara, CA, USA) was used for quantification. Nonadecanoic acid (C19:0) was used as an internal standard.
7
Fatty Acid Profiling of Prickly Pear
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Methanolic sulphuric acid and benzene were used for lipid methylation. Fatty acid profile of prickly pears cladodes extract and fruit juice were determined according to the method of Egan et al. (1981) using Gas Liquid Chromatography (GLC) analytical assay by Hewlett Packard (HP) 6890 GC, with Flame Ionization Detector (FID). The column phase was HP-88 (88%-Cyanopropyl-aryl-polysiloxane), 100 m, 0.25 mm ID, 0.2 µm film thickness, Nitrogen was used as a carrier gas with flow rate of 1 ml/min. The injector temperature was 220 o C. The results were expressed as fatty acids percentages. Finally, the calculated sums and ratios were; total monounsaturated fatty acids content (ΣMUFA), total polyunsaturated fatty acids content (ΣPUFA), total unsaturated fatty acids content (ΣUFA), total saturated fatty acids (ΣSFA), ΣPUFA/ΣSFA (P/S) ratio, ΣUFA/ΣSFA ratio and ω-6/ω-3 ratio.
8
Quantifying BDE Levels in Sediment, Worms, and Water
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The concentrations of the BDEs in the sediment, T. tubifex and water were determined by high-resolution gas chromatography coupled with high-resolution mass spectrometry (HRGC/HRMS).
The GC-HRMS was performed with a HP 6890 GC (Hewlett-Packard, Palo Alto, CA, USA) coupled to a Finnigan MAT 95XP (Finnigan, Bremen, Germany) high-resolution mass spectrometer. The GC separation was performed An aliquot (2 μL) of sample extract was injected into the GC system in pulsed splitless mode at 250 °C. The mass spectrometer operated in the electron impact ionization mode using selected ion monitoring (SIM) at a minimum resolution of 8000. The samples were analyzed for the BDE concentrations using the isotope-dilution or internal-standard method based on the US EPA 1614 protocol. In addition to daily sensitivity and relative response factor (RRF) checks, the mean RRF was regularly re-evaluated for each congener.
The GC-HRMS was performed with a HP 6890 GC (Hewlett-Packard, Palo Alto, CA, USA) coupled to a Finnigan MAT 95XP (Finnigan, Bremen, Germany) high-resolution mass spectrometer. The GC separation was performed An aliquot (2 μL) of sample extract was injected into the GC system in pulsed splitless mode at 250 °C. The mass spectrometer operated in the electron impact ionization mode using selected ion monitoring (SIM) at a minimum resolution of 8000. The samples were analyzed for the BDE concentrations using the isotope-dilution or internal-standard method based on the US EPA 1614 protocol. In addition to daily sensitivity and relative response factor (RRF) checks, the mean RRF was regularly re-evaluated for each congener.
9
GC-MS Analysis of Organic Compounds
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The GC-MS analysis of 4HMBA, CHR, AcA and EPX was carried out with a 5973 Hewlett Packard selective mass detector (quadrupole), source 70 eV, coupled to a HP 6890 GC fitted with a HP-5MS column (5 % phenylmethyl siloxane, 30 m 9 0.25 mm) with helium as carrier gas (1.0 mL min -1 ; constant flow). The oven was programmed as follows: 150 °C (0 min), 150-180 °C (3 °C min -1 ), 180 °C (2 min), 180-234 °C (1.5 °C min -1 ), 234 °C (2 min). Injection: 0.1 lL of a 10 % solution of the compound dissolved in methylene chloride. Injector and detector temperatures were maintained at 250 °C and 270 °C, respectively. Injection port was maintained at 250 °C, GC-MS interphase at 275 °C, ion source 230 °C, and MS Quad at 150 °C.
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