To examine the composition of the phase above the surface of the oil, 3.0 g of the sample was incubated at 40 °C for 20 min. Each batch consisted of three samples. HS-SPME analysis was carried out using a ternary fibre Divinylbenzene/Carboxen/Polydimethylsiloxane (DVB/CAR/PDMS). Extraction time: 10 min, temperature: 40 °C. Gas Chromatography was conducted using a Trace GC Ultra chromatograph coupled with a DSQ II mass spectrometer (Thermo Electron, Waltham, MA, USA) and equipped with a STABILWAX®-DA (30 m × 0.18 mm × 0.18 μm) column. The HS-SPME conditions were as follows: injection temperature: 50 °C, detector temperature: 240 °C, developing gas: He, pressure: 110 kPa, split: 50 cm3 min−1, temperature program: 50 °C (2 min), 10 °C min−1 (230 °C), and 230 °C (30 min). Each experiment was conducted in triplicate.
Trace gc ultra chromatograph
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
The Trace GC Ultra chromatograph is a gas chromatography system designed for chemical analysis. It is capable of separating and detecting a wide range of volatile and semi-volatile compounds. The instrument utilizes a capillary column and a variety of detectors to identify and quantify the components in a sample.
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Lab products found in correlation
Dsq 2 mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Rtx 1, by Restek (1 mentions)
Cross linked polyethylene glycol, by Agilent Technologies (1 mentions)
Pdms fiber, by Merck Group (1 mentions)
Itq 700, by Thermo Fisher Scientific (1 mentions)
Agilent 7890, by Agilent Technologies (1 mentions)
Ab204 s, by Mettler Toledo (1 mentions)
Thermomixer compact, by Eppendorf (1 mentions)
Ctc combipal autosampler, by CTC Analytics (1 mentions)
8 protocols using trace gc ultra chromatograph
1
Headspace Volatile Profiling of Oils
2
Clove Essential Oil Composition Analysis
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The clove buds (Syzygium aromaticum (L.) Merr. and Perry) commercial essential oil was purchased from Pollena Aroma S.A. (Warszawa, Poland). The oil’s composition was analyzed by gas chromatography-mass spectroscopy (GC-MS) using Trace GC Ultra chromatograph (Thermo Electron Corporation, Waltham, MA, USA) equipped with Rtx-1 (Restek, Bellefonte, PA, USA) non-polar capillary column (60 m × 0,25 mm; 0,25 μm film thickness) combined with DSQ II mass spectrometer (Thermo Electron Corporation, Waltham, MA, USA), according to the procedure described by Smigielski et al. [52 (link)]. The temperature was programmed as follows: 50–300 °C at 4 °C/min; injector (SSL) temperature 280 °C; the detector (FID) temperature 300 °C; carrier gas helium with constant pressure 300 kPa; split ratio 1:37. The mass spectrometer operating parameters: Ion source temperature 200 °C; ionization energy 70 eV (EI). The identification of the oil’s components was based on the comparison of their retention indices (RI), mass spectra (NIST and Wiley libraries) and the literature [53 ].
3
Fatty Acid Composition Analysis
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The extraction of total lipids was performed in duplicate according to the Folch et al. [1957] method, while the derivatisation and determination of total fatty acid composition were performed according to the AOAC -approved method 991.39 [AOAC, 2006] . Briefly, to the fat extracted (10 mg), 0.5 mL of 0.5 N KOH (methanol solution) was added and heated to 85°C, then 1 mL of 12% BF 3 (methanol solution) was added, and the mixture was reheated at 85°C. After cooling to room temperature, 1 mL of hexane and 5 mL of saturated NaCl solution were added. About 1 μL of the resulting sample was injected onto the column connected to the TRACE GC UL-TRA chromatograph (Thermo Electron Corporation, Milano, Italy) with a flame ionization detector (FID). Operating parameters were as follows: FID temperature 250ºC; dispenser temperature 220ºC; oven temperature 160ºC (3 min) to 210ºC (3ºC/min) (210ºC, 35 min). SUPELCOWAX10 column (30 m, 0.25 mm, 0.25 μm) was used and helium was applied as a carrier gas at a flow rate of 1 mL/min. Split flow was 10 mL/min. Individual fatty acid methyl esters were identified by comparison to the standards of a mixture of Supelco 37 component FAME Mix, and of CLA isomers.
4
Glycosidic Linkage Analysis by GC-MS
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Washed and dried gradient fractions were hydrolyzed with 2 N CF3COOH at 110 °C for 2 h, dried under nitrogen and reduced with NaBH4 (10 mg/mL in 1 N NH4OH/CH3CH2OH 1:1, v/v) at room temperature for 2 h. The reaction was stopped with few drops of acetic acid, dried under nitrogen, and acetylated with pyridine anhydrous/acetic anhydride (1:1, v/v) at room temperature overnight. After drying under nitrogen, petroleum ether (1 mL) was added and 1 µL of samples were analyzed by GC-MS. GC-MS analyses were performed using a Thermo TraceGCultra chromatograph equipped with an Inferno ZB5HT column (30 m × 0.25 mm) and connected to an ISQ single quadrupole mass <220 °C with split ratio of 20:1. Helium circulates at a constant flow rate of 1.2 mL.min−1 as carrier gas. The temperature separation program was: initial temperature at 100 °C and then increased until 300 °C at a rate of 20 °C min−1, followed by 3 min at 300 °C.
5
Quantification of Volatile Compounds in Food
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The quantification of volatile compounds was performed following the protocol defined previously (39 (link)). Extraction was performed using headspace solid-phase microextraction sampling (SPME) with polydimethylsiloxane (PDMS) fibers (Supelco; Sigma-Aldrich, Barcelona, Spain). Aroma compounds were separated by gas chromatography using a TRACE GC ULTRA chromatograph (Thermo Fisher Scientific, Waltham, MA) equipped with a flame ionization detector (FID). The column used for separation was an HP-INNOWAX 30-m by 0.25-mm capillary column coated with a 0.25-mm layer of cross-linked polyethylene glycol (Agilent Technologies, CA). Helium was the carrier gas (flow rate, 1 mL/min). The oven temperature program was 5 min at 60°C, 5 min at 190°C, 20 min at 250°C, and 2 min at 250°C. The detector temperature was 280°C, and the injector temperature was 220°C under splitless conditions. 2-Heptanone (0.05% [wt/vol]) was used as an internal standard. The volatile compounds were identified by the retention time for the reference compounds. Quantification of volatile compounds was performed using the calibration graphs of the corresponding standard volatile compounds.
6
Characterization of Equisetum canadensis FF
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The influence
of the UAE process
of the separation of E. canadensis functional
FF on the overall chemical properties was analyzed using a variety
of colorimetric and spectroscopic techniques, described previously.26 (link) All colorimetric assays were performed with
a SPECTROStar UV–vis spectrophotometer (BMG LABTECH, Germany).
The contents of saccharides, uronic acids, and total phenolics were
estimated by the phenol–sulfuric, m-hydroxybiphenyl,
and Folin–Ciocalteu assay, respectively.25 (link) The monosaccharide composition was determined by the gas
chromatography-mass spectrometry (GC-MS) technique (Trace GC Ultra
chromatograph with an ion trap detector ITQ 700, Thermo Scientific,
equipped with an Agilent HP-88 column (0.25 mm × 30 m), where
the mixture of the neutral monosaccharides was obtained from the corresponding
bioproducts after hydrolysis with TFA (trifluoroacetic acid, 2 mol/L),
120 °C, 5 h) and derived to obtain the volatile alditol acetate
forms, prior to the analysis (seeText S4 , Supporting Information).25 (link) The separation
of the horseweed FF obtained through the optimized UAE procedure was
conducted with the GPC technique on a glass column (15 mm × 1500
mm) packed with a Sephacryl 300 HR gel (Sigma-Aldrich, Germany) with
aqueous 0.1 mol/L NaOH eluent, followed by determination of the molecular
weights of the obtained fractions.
of the UAE process
of the separation of E. canadensis functional
FF on the overall chemical properties was analyzed using a variety
of colorimetric and spectroscopic techniques, described previously.26 (link) All colorimetric assays were performed with
a SPECTROStar UV–vis spectrophotometer (BMG LABTECH, Germany).
The contents of saccharides, uronic acids, and total phenolics were
estimated by the phenol–sulfuric, m-hydroxybiphenyl,
and Folin–Ciocalteu assay, respectively.25 (link) The monosaccharide composition was determined by the gas
chromatography-mass spectrometry (GC-MS) technique (Trace GC Ultra
chromatograph with an ion trap detector ITQ 700, Thermo Scientific,
equipped with an Agilent HP-88 column (0.25 mm × 30 m), where
the mixture of the neutral monosaccharides was obtained from the corresponding
bioproducts after hydrolysis with TFA (trifluoroacetic acid, 2 mol/L),
120 °C, 5 h) and derived to obtain the volatile alditol acetate
forms, prior to the analysis (see
of the horseweed FF obtained through the optimized UAE procedure was
conducted with the GPC technique on a glass column (15 mm × 1500
mm) packed with a Sephacryl 300 HR gel (Sigma-Aldrich, Germany) with
aqueous 0.1 mol/L NaOH eluent, followed by determination of the molecular
weights of the obtained fractions.
7
GC-MS Analysis of Organic Compounds
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A gas chromatograph, Agilent 7890, coupled to a triple quadrupole mass spectrometer, Agilent 7000 Series Triple Quad GC/MS (Agilent Technologies, Wilmington, DE, USA) operating at 70 eV was employed. The GC was fitted with a split/splitless injector and a DB-5 MS capillary column (cross-linked 5% phenyl-methyl siloxane, 30 m x 0.25 mm i.d., 0.25 µm coating). In addition, a Trace GC Ultra chromatograph from Thermo (Bremen, Germany) equipped with a GC Triplus Autosampler, split/splitless inyector, a GC-Isolink interface and a Conflo IV universal interface with a Ni/CuO/Pt combustion reactor set a 1000°C was employed coupled to a Delta V advantage sector field mass-spectrometer (Thermo). An analytical balance model AB204-S (Mettler Toledo, Zurich, Switzerland) was used for the gravimetric preparation of all solutions. A centrifuge 5810R D from Eppendorf (Hamburg, Germany) was used to remove debris. A centrifugal vacuum concentrator from Genevac (Sulflok, UK) was employed for sample evaporation. A thermomixer compact from Eppendorf (Hamburg, Germany) was used to control the temperature of the derivatization reactions.
8
Metabolite Profiling of Biological Samples
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Ten biological replicates were prepared for each experimental condition, as described in Khodayari et al. (2013) (link). The samples were homogenized in 600 µL of methanol-chloroform (2:1) using a bead-beating device (Retsch TM MM301, RetschGbmH, Haan, Germany). A volume of 400 µL of ice-cold ultrapure water was added to each sample, before centrifugation at 4,000 g for 10 min at 4 °C. Then, aliquots (300 µL) of the upper aqueous phase containing polar metabolites were transferred to microtubes and vacuum-dried. Following derivatization (see Khodayari et al., 2013 (link) for the detailed procedure), metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS), which included a CTC CombiPal autosampler (CTC Analytics AG, Zwingen, Switzerland), a Trace GC Ultra chromatograph, and a Trace DSQII quadruple mass spectrometer (Thermo Fischer Scientific Inc., Waltham, MA, USA) (Khodayari et al., 2013) (link). Peaks were annotated using both mass spectra (two specific ions), and retention times. Calibration curves were set using standards consisting of 57 pure reference compounds most often quantified in insects with this equipement. Metabolite levels were quantified using XCalibur v2.0.7 software (Thermo Fisher Scientific Inc., Waltham, MA, USA).
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