The concentration of SCFAs (acetate, propionate, and butyrate) was quantified using fecal samples collected at weeks 17, 23, and 32. Approximately 20–50 mg of feces was extracted with 500 µL of 70% (v/v) ethanol and 0.1% (v/v) trifluoroacetic acid (TFA) solution spiked with an internal standard (4-methyl valeric acid) at a final concentration of 100 ppm. The solution was mixed thoroughly, then centrifuged at 14,000 x g at 4°C for 30 minutes to pellet the fecal material. The top 200 µL was removed and analyzed using a Shimadzu GC-17A gas chromatograph with a flame ionization detector (GC-FID, Shimadzu GC-17A). Samples were separated on a 30 m x 0.25 × 0.5 µm i.d. HP-INNOWax fused silica column (Hewlett-Packard, Australia) as per the manufacturer’s instructions. GC-FID analysis for each sample was performed in three technical replicates (n = 450). All measurements were normalized for the weight of fecal samples used for SCFA quantification.
Gc 17a
Manufactured by Shimadzu
Sourced in Japan, Italy, Australia, United States
The GC-17A is a gas chromatograph manufactured by Shimadzu. It is designed for the separation and analysis of complex organic compounds in a variety of samples. The GC-17A utilizes a flame ionization detector (FID) to quantify the separated compounds.
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Lab products found in correlation
Qp5050a, by Shimadzu (8 mentions)
Rxi 5 ms, by Agilent Technologies (6 mentions)
Tq8040, by Shimadzu (6 mentions)
Gcms tq8040, by Shimadzu (5 mentions)
Gcms qp5050a, by Shimadzu (5 mentions)
Gc 2010, by Shimadzu (4 mentions)
Omegawax 250, by Merck Group (4 mentions)
Hp innowax fused silica column, by Hewlett-Packard (3 mentions)
Gcms qp2010, by Shimadzu (3 mentions)
37 component fame mix, by Merck Group (3 mentions)
Supelco 37 component fame mix, by Merck Group (3 mentions)
Gcms qp5000, by Shimadzu (3 mentions)
Osmomat 030, by Gonotec (2 mentions)
20 hete, by Merck Group (2 mentions)
Aoc 20i, by Shimadzu (2 mentions)
Sptm 2560, by Merck Group (2 mentions)
Sac 5 capillary column, by Merck Group (2 mentions)
Stg 1 nc, by ZenBio (2 mentions)
Luna c18 column, by Phenomenex (2 mentions)
Alltech 3300, by Büchi (2 mentions)
171 protocols using gc 17a
1
Fecal SCFA Quantification by GC-FID
2
Characterization of Cineole Metabolism
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General reagents, 1,8-cineole 1 and 1,4-cineole 2 were from Sigma-Aldrich. Buffer components, NADPH, and isopropyl--D-thiogalactopyranoside (IPTG) were from Astral Scientific (Australia) or VWR. Production and purification of full-length CYP102A1 variants for in vitro use were carried out as described previously [81, 92] . UV/Vis spectroscopy was performed on an Agilent Cary 60 spectrophotometer with temperature control at 30 C. Gas chromatography mass spectrometry (GC-MS) analyses were carried out on a Shimadzu GC-2010 coupled to a GC-MS-QP2010S detector or a GC-17A coupled to a QP5050A MS detector. Both systems used a DB-5 MS fused silica column (30 m x 0.25 mm, 0.25 µm) and helium as the carrier gas. The GC retention times are given in the figure legends and the methods in the supporting information. Additional GC analysis and enantioselective chromatography were performed on a Shimadzu Tracera GC coupled to Barrier discharge Ionization Detector (BID) detector using a Supelcowax column (30 m x 0.32 mm x 0.25 um) and a RT ® -BDEXse chiral silica column (Restek; 30 m x 0.32 mm x 0.25 um), respectively.
Enantioselective GC analysis of the cineole metabolites was performed on Cyclodextrin-B column (25 m × 0.25 mm) on a Shimadzu GC-17A equipped with an FID.
Enantioselective GC analysis of the cineole metabolites was performed on Cyclodextrin-B column (25 m × 0.25 mm) on a Shimadzu GC-17A equipped with an FID.
3
Plasma Fatty Acid Profiling in Clinical Evaluation
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Medical history, physical examination, and biochemical examination findings were examined. Medical history interrogated factors such as age, sex, case history, drinking habits, and history of prescribed medication. Patients were wearing light gowns and no shoes when height and body were measured. Venous blood samples were collected on the morning of the second day of hospitalization after a 12-h fast. Plasma fatty acid concentrations were measured by gas chromatography. Plasma was mixed with derivatizing reagent (KOKUSAN CHEMICAL Co., Ltd. Tokyo, Japan) and internal standard liquid, and was stirred to obtain methyl-esterified specimens. Then, NaOH and n-hexane were added to each methyl-esterified sample. The upper layer of the sample was separated after thorough mixing and centrifugation, and was measured using a gas chromatograph (GC-17A and GC-2010, SHIMADZU CORPORATION Co. Ltd., Kyoto, Japan) in SRL, Inc. (Tokyo, Japan). The biochemical data included alanine aminotransferase (ALT), gamma-glutamyl transpeptidase, creatinine (Cre), hemoglobin A1c (HbA1c), total cholesterol (TC), and triglyceride (TG) levels.
4
Fatty Acid Profiling of Avian Muscle
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To establish the fattening state of each bird, representative sub-samples of the pectoral muscle were pooled and homogenized; AOAC (Association of Official Analytic Chemists, Rockville, MD, USA) procedures were used to assess moisture, ether extract, protein, and ash content [20 ].
Fat was extracted according to the method suggested by Folch et al. [21 (link)], using a 2:1 chloroform/methanol (v/v) solution to determine the fatty acid profile. The fatty acids were then methylated using a KOH/methanol 2N solution [22 ] and analyzed via gas chromatography (Shimadzu GC-17A) using a silicone-glass capillary column (70% Cyanopropyl Polysilphenylene-siloxane BPX 70 produced by Thermo Scientific; length = 60 m, internal diameter = 0.25 mm, and film thickness = 0.25 µm). The starting temperature was 135 °C for 7 min; then, it was increased by 4 °C/min up to 210 °C. Samples of each concentrate mixture were used for fatty acid analysis according to the method described above to acquire muscle tissue fatty acid profiles. Fatty acids were expressed as a percentage (wt/wt) of total methylated fatty acids.
Fat was extracted according to the method suggested by Folch et al. [21 (link)], using a 2:1 chloroform/methanol (v/v) solution to determine the fatty acid profile. The fatty acids were then methylated using a KOH/methanol 2N solution [22 ] and analyzed via gas chromatography (Shimadzu GC-17A) using a silicone-glass capillary column (70% Cyanopropyl Polysilphenylene-siloxane BPX 70 produced by Thermo Scientific; length = 60 m, internal diameter = 0.25 mm, and film thickness = 0.25 µm). The starting temperature was 135 °C for 7 min; then, it was increased by 4 °C/min up to 210 °C. Samples of each concentrate mixture were used for fatty acid analysis according to the method described above to acquire muscle tissue fatty acid profiles. Fatty acids were expressed as a percentage (wt/wt) of total methylated fatty acids.
5
Measurement of Urinary 20-HETE
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Urine osmolality was determined by freezing-point depression using a semi-micro osmometer (Osmomat 030, Gonotec, Germany) and sodium concentrations by flame photometer (Jenway PFP7, Essex, UK). 20-HETE concentration in urine samples was measured by gas chromatography (Shimadzu GC-17A, Shimadzu, Japan) using own calibration standards prepared from synthetic 20-HETE (Sigma, USA).
6
Trace Ion and Gas Analyses in Bioreactors
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The concentration of NO3−, NO2−, PO43−, NH4+ and SO42− in the anodic and cathodic liquid stream were determined by an Ion Chromatograph (DIONEX-500 fitted with GP50 Gradient pump and CD20 conductivity detector) with IonPac CS12A cation and IonPac AS9-HC anion column. In those measurements, samples were first filtered through a 0.2-μm pore sized membrane before analysis. Acetate was analyzed using a gas chromatograph (Shimadzu, AOC-20i) equipped with a FIT detector and a 25 m × 0.32 mm × 0.5 μm HP-FFAP column. Samples were also filtered through a 0.2 μm pore sized membrane and acidified using formic acid before analysis. Produced N2 gas analyses were performed using a gas chromatograph (GC-17A, Shimadzu) with charlston 80/100 porapak column using Helium gas as carrier. Total nitrogen was measured using a Shimadzu TNM-1 unit coupled with a TOC-V analyzer. In both case, samples were pre-filtered through a 0.2 μm pore sized membrane.
7
Ethylene Production in Fruit Abscission
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Cumulative fruit abscission rate and relative fruit abscission rate were calculated according to our previous method (Kuang et al., 2012 (link)). Ethylene production was measured according to the method described by Yan et al. (2011) (link) with some modifications. Two fruit from each treatment on each tree were collected and enclosed in a 25 mL airtight syringe equipped with a rubber piston for 2 h at 25°C. Air within the syringe was forced into an airtight container filled with saturated salt water with a needle inserted to allow replacement. After all the air samples were collected in the experiment, 1 mL air sample was then withdrawn from the headspace of the container with a syringe and injected into a GC-17A gas chromatograph (Shimadzu, Kyoto, Japan) fitted with a flame ionization detector and an activated alumina column (200 cm × 0.3 cm). The injector temperature was 120°C; the column temperature was kept at 60°C and the detector temperature at 60°C. Helium was used as carrier gas at a flow rate of 30 mL⋅min-1. The ethylene production rate was expressed as microliters of C2H4 kg-1⋅h-1.
8
Extraction and Analysis of Silkworm Larvae Fatty Acids
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The total fatty acids in unfermented, and fermented silkworm larvae were extracted according to the Folch, Lees, & Stanley [20 (link)] method with some modifications. Briefly, 1 g of fermented silkworm larvae powder were extracted at 37 °C for 30 min using chloroform: methanol 2:1 (v/v) containing butylated hydroxyl toluene to inhibit the oxidation of fatty acids. Following centrifugation (3000×g, 15 min), the part of chloroform was concentrated in rotary vacuum evaporator to remove chloroform solvent. Extracted fatty acids were trans methylated to the fatty acid methyl esters by adding 5 mL of methanolic HCl (5:1, v/v) at 65 °C for 3 h. The fatty acid methyl esters were analyzed using gas chromatography (GC-17A, Shimadzu Co., Kyoto, Japan), equipped with an omegawax® capillary GC column (30 m × 0.25 mm × df 0.25 μm). Helium was the carrier gas, and the column flow rate was 1 mL/min. One μL of sample was injected in splitless mode into an inlet held at 250 °C, and the oven program was 180 °C. The data are expressed as mg/g of detecting total analyzed fatty acids.
9
Spectrophotometric Analysis of Fatty Acid Hydroperoxide Metabolism
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The UV spectra of the reaction mixtures were scanned during the incubations of CYP440A18 with fatty acid hydroperoxides with Varian Cary 50 spectrophotometer. Alternatively, the UV spectra of products were recorded online during the HPLC separations using an SPD-M20A diode array detector (Shimadzu, Japan). Products (Me esters or Me/TMS derivatives) were analyzed by GC–MS as described previously [21 (link)]. GC–MS analyses were performed using a Shimadzu QP5050A mass spectrometer connected to a Shimadzu GC-17A gas chromatograph equipped with an MDN-5S (5% phenyl 95% methylpolysiloxane) fused capillary column (length, 30 m; ID 0.25 mm; film thickness, 0.25 µm). Helium at a flow rate of 30 cm/s was used as the carrier gas. Injections were made in the split mode using an initial column temperature of 120 °C, injector temperature 230 °C. The column temperature was raised at 10 °C/min until it reached 240 °C. Electron impact ionization (70 eV) was used.
10
Nitrogenase Activity Measurement by GC
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The nitrogenase activity of an isolated strain was determined using gas chromatography (Shimadzu GC-17A) ethylene and the acetylene reduction technique (Baldani et al., 1986 ). The nitrogenase activity was calculated using the method described by Yao et al. (2004) . The experiments were carried out in triplicate.
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