The analyses were performed by means of a helium GC (Thermo Finnigan, Dreieich, Germany) using DB-FFAP (30 m × 0.32 mm fused silica capillary, 0.25 μm; Agilent J&W GC Columns, USA) and DB-5 (30 m × 0.32 mm fused silica capillary, 0.25 μm; Agilent J&W GC Columns, USA). The injection volume was 2 μL and the carrier gas flow was 2.2 mL min−1. The initial temperature for both columns was 40°C and was held for 2 min. Then, the temperature was raised at 8°C min−1 till 250°C and held for 10 min (DB-5), or to 245°C and held for 8 min (DB-FFAP). The temperature of both sniffing port and FID was set to 270°C. The GC effluent was split 1:1 between the sniffing port and the detector (FID or MS; DSQ, Thermo Finnigan, Dreieich, Germany). The linear retention index (RI) of each compound was calculated according to (Den Dool and Kratz, 1963 (link)). Mass spectra were obtained in the electron impact (EI) mode using the following conditions: 70 eV ionization energy, mass range 35 to 249 m/z, scan rate 500 amu/s and a source temperature of 200°C. Three experts were recruited from the sensory panel of Fraunhofer IVV to perform the GC-O analyses. They had experience in recognizing the odor-active compounds in matrices similar to fish feed, i.e., aquaculture water and fish samples from our previous studies.
Agilent j w gc column
Manufactured by Agilent Technologies
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Agilent J&W GC Columns are high-performance capillary columns designed for gas chromatography applications. They provide consistent and reliable separation of a wide range of analytes, enabling accurate and reproducible results.
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Lab products found in correlation
Agilent 7890a, by Agilent Technologies (2 mentions)
Db ffap, by Agilent Technologies (1 mentions)
Agilent 7890a gc, by Agilent Technologies (1 mentions)
Agilent 5975c inert xl msd, by Agilent Technologies (1 mentions)
Multipurpose sampler, by Gerstel (1 mentions)
Cp 3800 gc, by Agilent Technologies (1 mentions)
Agilent 6850, by Agilent Technologies (1 mentions)
Agilent 7890a gc system, by Agilent Technologies (1 mentions)
9 protocols using agilent j w gc column
1
Comprehensive GC-O Analysis of Fish Feed
2
GC-MS Analysis of Nigella sativa Seed Oil
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The chemical composition of the seed oil isolated from Nigella sativa L., varieties utilized in the study was determined through GC-MS analysis following the AOCS Method CE 1-62 and Cupido et al. (2022) with minor modifications. The GC-MS profiling was done using Agilent (GC-7820 A, MS-5975; Agilent Technologies, Santa Clara, CA, USA) equipped with an HP 5 (universal column; 30 m × 0.325 mm, film thickness 0.25 μm) Agilent J&W GC column with an autosampler. A sample of 1 ml was used with an autosampler. Helium was used as the carrier gas at a flow rate of 1.0 ml/min. The column temperature was programmed from 50 °C to 280 °C with an equilibrium time of 3 min, held for 30 min. Injector temperatures were set at 250 °C. The biochemical constituents were identified by a comparison of their retention indices and their identification was confirmed by computer matching of their mass spectral fragmentation patterns of compounds in the NIST-MS library and published mass spectra with the help of Chemtation software (Agilent Technologies, Palo Alto, CA, USA).
3
Metabolite Derivatization and GC-MS Analysis
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Metabolite derivatization was performed by using a multi-purpose sampler (GERSTEL, Germany). Dried samples were dissolved in 15 μl pyridine, containing 20 mg/ml methoxyamine hydrochloride, at 40°C for 60 min under shaking. After adding 15 μl N-methyl-N-trimethylsilyl-triflouroacetamide (MSTFA) samples were incubated at 40°C for 30 min under continuous shaking.
GC-MS analysis was performed by using an Agilent 7890A GC coupled to an Agilent 5975C inert XL MSD (Agilent Technologies, Germany). A sample volume of 1 μl was injected into a Split/Splitless inlet operating in splitless mode at 270°C. The gas chromatograph was equipped with a 30 m DB-35MS capillary column + 5 m DuraGuard capillary in front of the analytical column (Agilent J&W GC Column).
GC-MS analysis was performed by using an Agilent 7890A GC coupled to an Agilent 5975C inert XL MSD (Agilent Technologies, Germany). A sample volume of 1 μl was injected into a Split/Splitless inlet operating in splitless mode at 270°C. The gas chromatograph was equipped with a 30 m DB-35MS capillary column + 5 m DuraGuard capillary in front of the analytical column (Agilent J&W GC Column).
4
GC/MS Analysis of Chemical Compounds
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GC/MS analysis was performed using a Varian CP-3800 GC equipped with an electronic flow control (Varian Inc., Palo Alto, CA, USA). A fused silica methyl-deactivated capillary column (internal diameter: 10 m × 0.25 mm) was used as a guard column connected to a VF-5 ms (internal diameter: 30 m, 0.25 mm and film thickness: 0.25 μm) analytical column (Cat. Number CP9013, Agilent J&W GC Columns, Agilent Technologies, Cernusco sul Naviglio, Italy). The initial column temperature was set to 40°C for 5 min and then increased at 20°C/min to 220°C, which was maintained for 1 min (total, 15 min). The injector (250°C) was set in the split mode (10 : 1), and helium at a flow rate of 1.2 mL/min was used as the carrier. Ionization was performed using an ion-trap Saturn 2200 series MS detector operating in the electron impact mode.
5
Quantification of Fecal Short-Chain Fatty Acids
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Fecal samples (0.5 g) and distilled water containing 3 mmol/L of 2-ethylbutyric acid (as internal standard) and 0.5 mmol/L of H2SO4 were homogenized. 2.5 mL of the homogenate was vacuum distilled according to the method of Zijlstra et al.47 (link) and modified by Høverstad et al.48 (link). The distillate was analyzed with gas chromatography (Agilent 6850; Agilent, CA, USA) using a capillary column (serial no. USE400311H, Agilent J&W GC columns; Agilent, CA, USA) and quantified while using internal standardization. Flame ionization detection was employed. The total amount of all SCFAs and the amount of acetic, propionic, butyric, isobutyric, valeric, isovaleric, caproic, and isocaproic acids expressed in mmol/Kg wet weight were measured.
6
Nitrogenase Activity Quantification Protocol
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Nitrogenase assays were conducted in serum vials (total volume of 27 mL) containing 10 mL of the cell suspension. Sample vials were sealed and flushed with argon gas at room temperature for 5 min. The assay was initiated using an injection of 1 mL of acetylene gas (99.6%, gas cylinder of 5.0 kg) passed through an oil bubbler connected to a Schlenk line. The reaction was performed at 40°C for 1 h at 8 μmol m−2 s−1 illumination and then terminated by the addition of 0.3 mL of 30% trichloroacetic acid. Nitrogenase activity was routinely assessed based on the acetylene reduction reaction (8 (link)). Ethylene was measured by a gas chromatograph (7890B; Agilent Technologies, Santa Clara, CA, USA) with flame ionization detection (FID) fit with a GS-GASPRO capillary column (0.32 mm×30 m, Agilent J&W GC columns; Agilent Technologies). Helium was used as the carrier gas at a constant flow of 25 mL min−1; the temperatures of the oven, detection, and injection port were 40, 300, and 46.8°C, respectively. The amount of ethylene produced by each sample was calculated based on the standard curve obtained from different concentrations of ethylene gas (GL Sciences, Tokyo, Japan). The specific activity of nitrogenase was expressed as unit mg−1 of Chl a, where 1 unit is equal to the production of 1 nmol of ethylene h−1.
7
Quantifying Fecal Short-Chain Fatty Acids
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The method used to determine faecal SCFA levels has been described in detail previously[25 (link)]. Briefly, the faecal samples were homogenized with a solution containing 3 mmol/L 2-ethylbutyric acid and 0.5 mmol/L H2SO4. The homogenate was vacuum distilled, and the SCFA levels were determined by gas chromatography (Agilent 7890 A, Agilent, CA, United States) using a capillary column (serial no. USE400345H, Agilent J&W GC columns, Agilent) and flame ionization[35 (link),36 (link)] levels of total SCFAs, acetic, propionic, iso-butyric, n-butyric, iso-valeric, n-valeric acid, isocapronic and n-capronic acids, were determined and were expressed in units of mmol/kg wet weight.
8
GC-MS Analysis of Volatile Compounds
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The sample was injected using an injector (Curie Point Injector, Japan Analytical Industry Co., Ltd, Japan) to apply the adsorbed volatile components from the collection tube to the GC-MS system. A splitless injection method was used. Gas chromatography was performed by an Agilent GC system (7890A, Agilent CA. USA), and mass spectrometry was performed by a mass selective detector (MSD) (5975C, Agilent CA. USA). The GC capillary column (DB-5MS + DG, 30 m x 0.25 mm I.D. Agilent J&W GC Columns, Agilent CA. USA) was used for the analysis, and helium at a constant ow rate of 1 mL/min was used as the carrier gas. The GC oven was heated from 60ºC (initial time: 1 min) to 200ºC using a 5ºC/min temperature gradient and kept at 200ºC for 5 min, then heated to 325ºC at 60ºC/min and kept at 325ºC for 2 min. The measurement time was 38 minutes. Data analysis was performed using multivariate analysis software (Mass Pro ler Professional (MPP) Agilent CA, USA). The latest version of the National Institute of Standards and Technology (NIST) 2019 edition database [11] 267,376 spectra of common compounds, Automatic mass spectrometry and identi cation (AMDIS) [12] , NIST was used as a reference. Gender, age, relationship with in ammatory markers (WBC, CRP, erythrocyte sedimentation rate (ESR)), and changes before and after treatment were examined.
9
Fecal SCFA Quantification Protocol
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The fecal samples were weighed and homogenized with a solution containing 3 mmol/L 2‐ethylbutyric acid and 0.5 mmol/L H2SO4. A sample (2.5 mL) of the homogenate was vacuum‐distilled, and the SCFA levels were determined by gas chromatography (Agilent 7890 A; Agilent) using a capillary column (serial no. USE400345H; Agilent J&W GC columns, Agilent).14 , 15 Flame ionization was used to determine the levels of total SCFAs, acetic, propionic, isobutyric, n‐butyric, isovaleric, n‐valeric acid, isocapronic, and n‐capronic acids, with the results expressed in units of mmol/kg wet weight.
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