One microliter of derivatized samples (third fraction) was injected into a gas chromatograph (Agilent 7890B; Agilent Technologies, Waldbronn, Germany) coupled to a triple quadrupole mass spectrometer (QQQ/MS; 7000 C Agilent Technologies, Waldbronn, Germany), equipped with a high sensitivity electronic impact source (EI) operating in positive mode. The injection was performed in splitless mode. Front inlet temperature was kept at 250 °C, transfer line and ion-source temperatures were 250 and 230 °C, respectively. Septum purge flow was fixed at 3 mL/min. The purge flow to the split vent was operating at 80 mL/min for 1 min and gas saver mode was set to 15 mL/min after 5 min. Helium gas flowed through the column (HP-5MS, 30 m × 0.25 mm, i.d. 0.25 mm, d.f. J&WScientific, Agilent Technologies Inc.) at 1 mL/min. The column temperature was held at 60 °C for 1 min, raised to 210 °C (10 °C/min), then to 230 °C (5 °C/min), to finally reach 325 °C (15 °C/min), and held for 5 min. The collision gas was nitrogen.
J w scientific
Manufactured by Agilent Technologies
Sourced in Israel
J&W Scientific is a product line of laboratory equipment manufactured by Agilent Technologies. The core function of J&W Scientific products is to provide reliable and precise analytical solutions for various scientific applications. These products are designed to meet the needs of researchers, analysts, and laboratory professionals across diverse industries.
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Lab products found in correlation
4 protocols using j w scientific
1
Gas Chromatography-Mass Spectrometry Analysis
2
Proteomic Analysis of Amebic Proteins
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The proteins in each gel slice were reduced with 2.8 mM DTT (60°C for 30 minutes), modified with 8.8 mM iodoacetamide in 100mM ammonium bicarbonate in the dark at room temperature for 30 minutes, and digested overnight in 10% acetonitrile and 10 mM ammonium bicarbonate with modified trypsin (Promega-Biological Industries, Israel) at 37°C. The resulting peptides were resolved by reverse-phase chromatography on 0.075 x 200-mm fused silica capillaries (J&W Scientific, Agilent Technologies, Israel) packed with Reprosil reversed phase material (Dr. Maisch GmbH, Germany). The peptides were eluted at flow rates of 0.25 μl/min on linear gradients of 7–40% acetonitrile in 0.1% formic acid for 95 minutes followed by eight minutes at 95% acetonitrile in 0.1% formic acid. MS was done by an ion-trap mass spectrometer (Orbitrap, Thermo) in a positive mode using a repetitively full MS scan followed by collision-induced dissociation (CID) of the seven most dominant ions selected from the first MS scan. The MS data was analyzed using the Proteome Discoverer software version 1.3 which searches the Ameba section of the NCBI-NR database and the decoy databases (in order to determine the false discovery rate (FDR)) using the Sequest and the Mascot search engines.
3
Fatty Acid Methyl Esters Analysis
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Fatty acid methyl esters were analyzed on a Varian 3900 gas chromatograph (GC) equipped with a DB-FFAP 15 m × 0.10 mm i.d. × 0.10 μm film thickness, nitroterephthalic acid modified, polyethylene glycol, capillary column (J&W Scientific from Agilent Technologies, Mississauga, ON) with hydrogen as the carrier gas. Samples (1 µL) were introduced by a CTC Combi PAL (CTC Analytics AG, Zwingen, Switzerland) into the injector heated to 1 3 250 °C with a split ratio of 100:1. The initial temperature was 150 °C with a 0.25-min hold followed by a 35 °C/min ramp to 200 °C, an 8 °C/min ramp to 225 °C with a 3.2min hold and then an 80 °C/min ramp up to 245 °C with a 15-min hold at the end [26] . The flame ionization detector temperature was 300 °C with air and nitrogen make-up gas flow rates of 300 and 25 mL/min, respectively, and a sampling frequency of 80 Hz. Peaks were identified by retention times through comparison to an external mixed standard sample (GLC-462, Nu Chek Prep Inc., Elysian, MN, USA).
4
Comprehensive Fatty Acid Profiling of Dietary Oils
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Fatty acid analysis of arachidonic acid-rich dietary oil and vehicle oil AA-rich dietary oil and the vehicle oil (n 3) were diluted with a methanol-hexane vehicle containing butylhydroxytoluene as an antioxidant and docosatrienoic acid methyl ester (C22 : 3n-3 methyl ester) as an internal standard. After adding acetyl chloride, the mixture was heated at 100°C for 60 min for extraction and methylation of lipids. The samples were shaken with potassium carbonate solution. Fatty acid methyl esters in the hexane layer were determined using Agilent 7890A gas chromatograph (Agilent Technologies) with a split injector (Agilent 7693 automatic liquid sampler; Agilent Technologies), and were detected using a flame ionisation detector (Agilent Technologies). The GC column used was a DB-FFAP 15 m × 0•10 mm i.d. with 0•10 µm film thickness (J&W Scientific from Agilent Technologies). The oven temperature programme was initiated at 150°C with a 0•25-min hold, and then ramped up at 35°C/min to 200°C, then at 8°C/min to 225°C with a 3•2-min hold and finally ramped up at 80°C/min to 248°C with a 14•7-min hold. The detector and injector temperatures were both set at 250°C.
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