Analysis of the methidathion pesticide was carried out with a Hewlett–Packard 6890 gas chromatograph equipped with an NPD Detector, on-colum injection port, and HP-5 column (5 % diphenyl copolymer/95 % dimethylpolysiloxane) (25 m × 0.32 mm ID, 0.52 μm film thickness)and temperature programming from 80 to 160 °C at 25 °C/min. 220–240 °C at 10 °C/min, 80 °C (3.00 min), 160 °C (2.00 min), 220 °C (10.00 min), 240 °C (8.80 min); Injector temperature 73–250 °C (180 °C min). The temperature of the detector was 300 °C. Carrier gaz (helium) flow rate, 2.6 mL/min; makeup gaz (nitrogen) flow rate, 10 mL/min; Air 60 ml/min; H2 3 mL/min. The injection volume was 1 μL.
6890 gas chromatograph
Manufactured by Hewlett-Packard
Sourced in United States
The Hewlett-Packard 6890 gas chromatograph is a laboratory instrument designed for the separation, identification, and quantification of chemical compounds in complex mixtures. It utilizes a capillary column and a detector to analyze the components of a sample as they are separated based on their physical and chemical properties.
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Lab products found in correlation
5973 mass spectrometer, by Agilent Technologies (2 mentions)
Bpx 70 column, by Hewlett-Packard (2 mentions)
Gas chromatograph, by Bruker (1 mentions)
Hp 5ms capillary column, by Hewlett-Packard (1 mentions)
Hp 5ms column, by Hewlett-Packard (1 mentions)
Selectfame column, by Agilent Technologies (1 mentions)
Fame standard, by Merck Group (1 mentions)
Heptadecanoic acid, by Merck Group (1 mentions)
Butylated hydroxytoluene, by Merck Group (1 mentions)
Acetyl chloride, by Merck Group (1 mentions)
Bpx70 column, by Agilent Technologies (1 mentions)
Sptm 2560 capillary column, by Merck Group (1 mentions)
5973n mass selective detector, by Hewlett-Packard (1 mentions)
5973a mass spectrometer, by Hewlett-Packard (1 mentions)
Gc ms, by Hewlett-Packard (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
20 protocols using 6890 gas chromatograph
1
Methidathion Pesticide Analysis by GC-NPD
2
Monosaccharide Linkage Analysis by GC-MS
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Alcohol-insoluble residue (AIR) was prepared and methylation analysis for both neutral and acidic monosaccharide linkage composition was performed following the procedure described by Pettolino et al. (2012) (link). Monosaccharide linkage analysis was performed on a Hewlett-Packard 6890 Gas Chromatograph with a Hewlett-Packard 5973 Mass Spectrometer (Agilent) equipped with a BPX70 column (25 m × 0.22 mm inner diameter, film = 0.25 μm, SGE).
3
GC-FID and GC-MS Analysis of Essential Oils
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GC analysis was carried out on a Bruker gas chromatograph with an FID detector and a BP-1 capillary column (30 m × 0.25 mm; film thickness 0.25 µm). The operating conditions were as follows: the carrier gas was helium with a flow rate ranging from 6–10 mL/min at 1 mL/min; the column temperature ranged from 50 to 280 °C at 5 °C/min; the injector and detector temperature was 280 °C; the volume of oil injected was 0.1 µL, with a split ratio of 1:50. The GC/MS analysis was performed on a Hewlett Packard 6890 MS selective detector coupled with a Hewlett Packard 6890 gas chromatograph equipped with a cross-linked 5% PHME siloxane HP-5MS capillary column (30 m × 0.25 mm; film thickness 0.25 µm) and operating under the same conditions as described above. The MS operating parameters were as follows: ionization potential 70 eV, ionization current 2A, ion source temperature 200 °C, and the column oven temperature was programmed from 50 to 325 v °C. The m/z was set at 40–600 and resolution 1000. The percentage of the samples and quantification of components were computed from the GC peak areas. Identification of the EO constituents was based on the literature and fragmentation patterns of compounds in the system database.
4
Trichothecene Intermediate Compound Detection
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We used GC-MS to detect other trichothecene intermediate compounds, e.g., trichodermol, trichodiene, isotrichodiol, 12,13-epoxytrichothec-9-ene (=EPT, trichothecene) (Cardoza et al., 2011 (link)), and other secondary metabolites also known to have antifungal activity, e.g., aspinolides. TP6.6 strain was grown in liquid yeast extract-peptone-dextrose (YEPD) cultures [5% glucose, 0.1% yeast extract, 0.1% peptone (Difco, Becton Dickinson)] (20 mL YEPD in 50 ml Erlenmeyer flasks) at 200 rpm and 28°C or 20°C, which are optimal and suboptimal temperatures, respectively, for trichothecene production. After 7 days, cultures were extracted with 3 mL of ethyl acetate. One μL of the extract was injected into a Hewlett Packard 6890 gas chromatograph fitted with a HP-5MS column (30 m, 0.25 mm, 0.25 µm) and a 5973-mass detector. The carrier gas was helium with a 20:1 split ratio and a 20 mL/min split flow. The column was held at 150°C for one minute following injection, heated to 280°C at 30°C/min and held for 3.6 min for a total run time of 9 min. Compound identifications were based on GC-MS comparisons with purified standards.
5
Plasma Fatty Acid Analysis Protocol
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Biochemical analysis was performed as described by Folch et al. [20] (link). Blood samples were collected in ethylenediaminetetraacetic acid (EDTA) tubes and kept on ice. Plasma for fatty acid analysis was separated by centrifugation at 3,000 rpm for 10 min. at 4°C and then frozen at −80°C until analysis. Lipids were extracted from 0.20 mL of plasma in chloroformmethanol (2:1) containing 10 mg/L of butylated hydroxytoluene (BHT) [20] (link). Thin-layer chromatography (Hewlett-Packard 6890 gas chromatograph; Minnesota, USA) was used to separate and evaporate plasma phospholipids until dry under nitrogen gas. Fatty acids were classified against authentic lipid standards obtained from Nu-Chek Prep, Inc. (Elysian, MN, USA). We evaluated the saturated fatty acids (SFAs) C14 (myristic acid), C15 (pentadecanoic acid), C16 (palmitic acid), C18 (stearic acid), C20 (arachidic acid), C22 (behenic acid), and C24 (lignoceric acid); the omega-3 FFAs C22:5n3 (EPA) and C22:6n3 (DHA); the omega-6 FFAs C18:2n6 (LA), C20:4n6 (AA), and C20:3n6 (homo-gamma-linolenic acid); and the omega-9 FFAs C18:1n9 (oleic acid), C22:1n9 (erucic acid), and C24:1n9 (nervonic acid) (Table 1 ).
6
Erythrocyte Omega-3 Fatty Acids Analysis
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Fasting blood samples were collected from participants by a nurse into 4-mL sodium citrate vacutainer tubes (BD Vacutainer®, Franklin Lakes, NJ) and centrifuged at 4°C (4000g for 10 min). After centrifugation, erythrocytes were collected with a disposable pasteur pipette and transferred into eppendorfs, which were stored in a − 80°C freezer until further analysis. Erythrocyte EPA and DHA were assessed using gas chromatography as described elsewhere (19 ). Briefly, erythrocyte lipids were extracted into chloroform–methanol, and fatty acid methyl esters (representing the erythrocyte fatty acids) were formed by heating the lipid extract with methanolic sulfuric acid. The fatty acid methyl esters were separated by gas chromatography on a Hewlett Packard 6890 gas chromatograph fitted with a BPX-70 column using the settings and run conditions described elsewhere (19 ). Fatty acid methyl esters were identified by comparison with run times of authentic standards. Data are expressed as weight % of total fatty acids. O3I was calculated by summing the percentages of EPA and DHA according to Harris and von Schacky (5 (link)).
7
Fatty Acid Profiling by GC-FID
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Approximately 2 mL of the lipid-containing chloroform phase was removed and derivatized with 0.5 mL of 0.5 M potassium hydroxide in methanol and 1 mL of boron trifluoride methylation by heating at 70 °C. Fatty acid methyl esters (FAME) were extracted into 4 mL of hexane and washed with 2 mL of distilled water. The hexane (organic) phase was then transferred to 2 mL vials. Then, the samples were analyzed by gas chromatography. The aforementioned analysis was carried out on a Hewlett-Packard 6890 gas chromatograph (Wilmington, DE, USA) equipped with a split/splitless injector and a flame ionization detector (FID). FAMEs were separated using a SelectFame column (50 m × 0.25 mm × 0.25 μm, Agilent Technologies, Santa Clara, CA, USA), identified by comparison with the available FAME standards (Supelco, Bellefonte, PA, USA). Fatty acid content was calculated by comparing the area of individual peaks with the area of the peak of the internal standard and recalculated on the basis of the sample weight Response factors (FID) for individual fatty acids compared to internal standards were taken as unity. The concentration of fatty acids was expressed quantitatively [41 (link)].
8
Analytes Extraction and GC Analysis
9
Maternal-Fetal Fatty Acid Profiling
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Fatty acid composition in maternal and foetal whole blood, and in lipids extracted from liver samples from offspring at 4 weeks of age, was determined by Gas Chromatography (GC) on a Hewlett-Packard 6890 gas chromatograph using methods that have previously been described in detail [22 (link),24 (link)]. Individual fatty acid content was calculated based on peak area and response factors normalised to total fatty acid content and expressed as a percentage of total fatty acids.
10
Plasma NEFA Fatty Acid Analysis
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NEFA were analysed as their FA methyl esters (FAMEs) using GC–MS, as described in detail elsewhere [10 (link), 14 (link)]. Direct transesterification of all classes of lipids was carried out in a one-step reaction [14 (link)]. In brief, lipids were extracted from plasma after addition of internal standard (heptadecanoic acid) using isopropyl alcohol–heptane–sulfuric acid (40:10:1) (Merck, Darmstadt, Germany) and 0.01% butylated hydroxytoluene (≥99%, B1378; Sigma-Aldrich, St Louis, MO, USA). Lipids were separated by thin-layer chromatography using heptane–diethylether–acetic acid (80:30:1) (Merck) as mobile phase. FAs were extracted from silica gel in benzol–methanol (1:4) (Merck) overnight and derivatised to their corresponding methyl esters by addition of acetyl chloride (Merck) and incubation at 100°C for 1 h. After addition of benzol (Merck) and centrifugation, the FAME-containing supernatant fraction was analysed on a Hewlett Packard 6890 gas chromatograph (Palo Alto, CA, USA) interfaced to a Hewlett Packard 5975 mass selective detector. Calibration curves of reference FAs were processed in parallel for quantification [10 (link)].
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