For gas chromatography–mass spectrometry (GC-MS) analysis, fatty acids were extracted as described by Ryu et al. [51 (link)] with the following modifications. A powdered freeze-dried seed sample (0.5 g) was extracted in 1 mL n-hexane for 4 h, then 0.1 mL of 2 N potassium hydroxide in methanol was added. After centrifugation for 5 min at 3000× g, the collected supernatant was filtered using a 0.45-μm syringe filter. The fatty acid composition was analyzed using a GC-MS (Plus-2010, Shimadzu, Japan) instrument equipped with a HP-88 capillary column (J&W Scientific, Folsom, CA, USA, 60 m × 0.25 mm × 0.25 m) under the following conditions: ionization voltage, 70 eV; mass scan range, 50–450 mass units; injector temperature, 230 °C; detector temperature, 230 °C; injection volume, 1 L; split ratio, 1:30; carrier gas, helium; and flow rate, 1.7 mL/min. The column temperature program specified an isothermal temperature of 40 °C for 5 min increasing to 180 °C at the rate of 5 °C/min, then a subsequent increase to 28 °C at the rate of 1 °C/min. We identified the substances present in the extracts in accordance with their retention time and with reference to a mass spectral database (NIST 62 Library).
2010 plus
Manufactured by Shimadzu
Sourced in Japan
The Shimadzu 2010 Plus is a high-performance liquid chromatograph (HPLC) designed for analytical and preparative applications. It features a high-pressure pump, a variable wavelength UV-Vis detector, and a temperature-controlled column compartment. The 2010 Plus is capable of delivering precise and reproducible solvent flow rates, enabling efficient separation and detection of a wide range of compounds.
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Lab products found in correlation
Hp 88 capillary column, by Agilent Technologies (2 mentions)
Rtx wax column, by Shimadzu (2 mentions)
Tq8040, by Shimadzu (2 mentions)
Rtx 5ms, by Shimadzu (1 mentions)
Carry 60, by Agilent Technologies (1 mentions)
Lc 20 ad t, by Shimadzu (1 mentions)
Db wax, by Agilent Technologies (1 mentions)
Finnigan delta plus xp, by Thermo Fisher Scientific (1 mentions)
Gcms qp2010 ultra, by Shimadzu (1 mentions)
Gcms tq8040, by Shimadzu (1 mentions)
Carbopack c, by Merck Group (1 mentions)
Atlantis dc18 column, by Waters Corporation (1 mentions)
Toc vcph cpn, by Shimadzu (1 mentions)
Ics 2100 ion chromatograph, by Thermo Fisher Scientific (1 mentions)
Agilent 1200, by Waters Corporation (1 mentions)
Uv 2600, by Shimadzu (1 mentions)
Ultra 2, by Agilent Technologies (1 mentions)
Cross linked capillary column, by Agilent Technologies (1 mentions)
Gc solution software, by Shimadzu (1 mentions)
400 mhz spectrometer, by Agilent Technologies (1 mentions)
25 protocols using 2010 plus
1
GC-MS Analysis of Seed Fatty Acids
2
GC-MS Analysis of Volatile Compounds in Rose Mutants
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The volatile compound compositions were analyzed using a GC-MS (Plus-2010, Shimadzu, Kyoto, Japan) equipped with a Rtx-5MS (30 m × 0.32 mm × 50 µm, Shimadzu, Kyoto, Japan) column. The carrier gas was 99.99% high-purity helium with a column flow rate of 1.37 mL/min. Sample injection was performed in splitless mode. The oven temperature was initially set at 40 °C, and was gradually increased to 300 °C at 5 °C/min with a final hold for 5 min. The mass spectrometry parameters included: Electron-impact ionization, 70 eV; ion source temperature, 230 °C; scan range, 40–500. The identification of each compound was performed using mass spectral libraries and Kovats retention indexes (RI). The GC-MS analysis detected volatile compounds in the rose mutants and those of the original cultivars, and compounds were tentatively identified based on a NIST library similarity index greater than 90%. The retention indices of all GC peaks were calculated with retention times of C7–C40 saturated alkane standards under the same chromatographic conditions. The RI of each compound on each column was calculated using the formula; y and z are carbon numbers of alkane standards, T(x) is the retention time of the compound, and n and n + 1 represent the retention times of the alkane standards.
3
Grain Oil Content Analysis
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Grain oil content was analyzed using the AOAC method as previously described (AOAC, 1990) (link). Using the Soxhlet extraction procedure, 5 g crushed seeds (80 mashed) were packed into a thimble and the oils were extracted with diethyl ether for 6 h. The lipid extracts of the grain and the whole plant were mixed with boron trifluoride (BF 3 )-methanol reagent (20%) and the fatty acids were converted into methyl ester derivatives. The methyl esters of the fatty acids were dissolved in chloroform (CHCl 3 ) and analyzed by gas chromatographymass spectrometry (Plus-2010, Shimadzu, Kyoto, Japan). The fatty acid composition of whole plants and grain was analyzed using a GC-MS instrument equipped with an HP-88 capillary column (J&W Scientific 60 m x 0.25 mm x 0.25 m) under the following conditions: ionization voltage, 70 eV; mass scan range, 35-450 mass units; injector temperature, 230°C; detector temperature, 230°C; inject volume, 1 µL; split ratio, 1:30; carrier gas, helium; and flow rate, 1.7 mL/min. The column temperature program specified an isothermal temperature of 40°C for 5 min followed by an increase to 180°C at a rate of 5°C/min and a subsequent increase to 28°C at a rate of 1°C/min. We identified the substances present in extracts by comparing their mass spectra against a database of GC-MS (Nist. 62 Library) spectra and by their retention indices.
4
Proximate, Mineral, Amino Acid Analysis of Food
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Proximate analysis was done following official methods of analysis of AOAC International8 . The moisture and dry matter contents were determined using a hot air oven by drying the sample at 105 °C for 8 hours. Ash content was determined at 600 °C. Nitrogen content was estimated by Kjeldahl method (Kelplus Classic, DX VA, Pelican Equipment) and crude protein was calculated by multiplying nitrogen percentage by 6.25. Lipid was determined by the solvent extraction method by Soxtec system (SOCS Plus, SCS-6, Pelican Equipment) using diethyl ether (boiling point, 40–60 °C) as a solvent. The crude fiber was determined by digesting the moisture and fat-free sample successively with dilute (1.25%) acid and alkali using Fibre cap (Foss tecator, Sweden). Nitrogen free extract (NFE) was determined as per the following formula
Fatty acid, amino acid, and mineral analysis were carried out following AOAC8 . Calcium, magnesium, chromium, manganese, iron, chromium analysis was performed in Absorption Spectrophotometer (Model 280 FS- Make Agilent Technology). Phosphorus content was estimated using UV-Vis Spectrophotometer (Model - Agilent Carry-60). Amino acids were analyzed using HPLC (Model - Shimadzu LC-20 AD/T)). Fatty acids were analyzed through gas chromatography (Model - Shimadzu 2010 Plus).
Fatty acid, amino acid, and mineral analysis were carried out following AOAC8 . Calcium, magnesium, chromium, manganese, iron, chromium analysis was performed in Absorption Spectrophotometer (Model 280 FS- Make Agilent Technology). Phosphorus content was estimated using UV-Vis Spectrophotometer (Model - Agilent Carry-60). Amino acids were analyzed using HPLC (Model - Shimadzu LC-20 AD/T)). Fatty acids were analyzed through gas chromatography (Model - Shimadzu 2010 Plus).
5
Fatty Acid Profiling of Bacterial Cells
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Bacterial suspensions treated with/without PAS were centrifuged to collect bacteria pellets. After the pellets were washed three times with PBS (0.01 M, pH 7.2), 40 mg of them were saponified and methylated and the fatty acid methyl esters were extracted according to the method reported by Wang, et al. [19 (link)]. One mL of the organic phase was transferred to a vial for subsequent Gas chromatographic (GC) analysis.
GC analysis of fatty acids was carried out with 37 kinds of fatty acid methyl esters as external standards. The GC spectrometer (2010 Plus, Shimadzu Co., Tokyo, Japan) was equipped with a capillary column (SPTM-2560, 100 m × 0.25 mm × 0.2 μm, Shimadzu Co., Tokyo, Japan). Helium was used as a carrier gas with a flow rate of 9.6 mL/min and in a split mode of 10:1 [17 (link)]. The initial pressure of the injector was 527 kPa, and the temperature of both the injector and flame ion detector (FID) was 260 °C. The temperature program was started at 100 °C, followed by an increase of 4 °C/min to 260 °C, and held for 30 min. The results were expressed as a ratio of total unsaturated fatty acids (area)/total saturated fatty acids (area).
GC analysis of fatty acids was carried out with 37 kinds of fatty acid methyl esters as external standards. The GC spectrometer (2010 Plus, Shimadzu Co., Tokyo, Japan) was equipped with a capillary column (SPTM-2560, 100 m × 0.25 mm × 0.2 μm, Shimadzu Co., Tokyo, Japan). Helium was used as a carrier gas with a flow rate of 9.6 mL/min and in a split mode of 10:1 [17 (link)]. The initial pressure of the injector was 527 kPa, and the temperature of both the injector and flame ion detector (FID) was 260 °C. The temperature program was started at 100 °C, followed by an increase of 4 °C/min to 260 °C, and held for 30 min. The results were expressed as a ratio of total unsaturated fatty acids (area)/total saturated fatty acids (area).
6
Rapid GC Analysis of Hexanol
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Reactions were analyzed on a Shimadzu 2010 Plus equipped with a flame ionization detector and a ZB-5 column (30 m, 0.25 µm, 0.32 mm, Agilent technologies). Sample aliquots of 1 μl were injected in split mode (split ratio 10:1). A rapid method for hexanol detection was used as an indicator for active enzymes in screening. Column temperature was increased from 90°C to 320°C at 30°C/min. For a more precise quantification of produced green leaf volatiles we applied a slower ramp. Initial temperature was 70°C, and the oven temperature was increased to 320°C at 20°C/min. Analytical standards for hexanal and hexanol, as well as internal standard tetradecane, were used to determine product concentrations. Quantitation was performed based on linear intrapolation of calibration curves with concentrations of standards ranging from 0.05 mM to 20 mM.
7
Fatty Acid Profiling by GC-FID
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For fatty acid profile analysis, approximately 0.1 g of ground samples and 2 ml of heptane were vortexed for 1 min, left standing for 16 h at 20°C and then centrifuged at 12,000 rpm for 5 min. The supernatant was transesterified adding 1 ml of methanol (4 N potassium hydroxide) and manually stirring for 1 min. The solution was dried with sodium sulfate, centrifugated at 12,000 rpm for 5 min. In total, 200 μl of the upper phase was diluted to 1 ml final volume, filtrated and injected into a gas chromatography equipment coupled with flame ionization detector (GC–FID, Shimadzu 2010 Plus, Japan). The column used was Agilent DB–WAX (30 m × 0.25 mm ID, 0.25 μm). The injection temperature was 250°C and the FID detector temperature was set at 300°C. The oven temperature was set at 13 min from 160 to 200°C and stay until 22 min, then the temperature increases to 240°C ending the program at 25 min. The sample injection volume was 1 μl, the mobile phase was hydrogen at 40 ml/min (split ratio at 180). Standard FAME (fatty acids methyl esters) was used for identification of the peaks and content was expressed as percentage of the total fatty acids.
8
Methane Stable Isotope Analysis Protocol
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Gas compositions were analysed by the same method as described above. δ3CCH4 was measured by coupling a Trace GC ultra gas chromatograph via a GC combustion interface (Thermo Surveyor) to a mass spectrometer (Thermo Finnigan Delta Plus XP).
Additional gas samples from gas emission spot 1 were analysed for δ13C and δ2H in CH4 at Imprint Analytics GmbH (Neutal, Austria) using a gas chromatograph (Shimadzu GC 2010Plus, Shimadzu Corp., Kyoto, Japan) coupled to a stable isotope ratio mass spectrometer (Nu Horizon; Nu Instruments Limited, Wrexham, UK). Gas samples were directly injected by an AOC5000 Autosampler (CTC Analytics, Zwingen, Switzerland) from the Labco Exetainers into the S/Sl Injector of the GC. The CH4 was separated from other gases in a Q-Plot GC column (Supelco, Bellefonte USA) at 35°C (isothermal). The separated analytes were identified in a quadrupole mass spectrometer (Shimadzu GCMS-QP2010Ultra). Methane was oxidized to CO2 for the δ13C analyses in an oxidation oven filled with oxidized Ni and Pt wires at a temperature of 1040°C. For the δ2H analysis, CH4 was pyrolysed at 1400°C to H2 in a ceramic tube (Hekatech, Wegberg, Germany). As reference material hexane vapour with a known δ13C and δ2H composition was used. δ13C is reported in ‰ VPDB, δ2H in ‰ SMOW.
Additional gas samples from gas emission spot 1 were analysed for δ13C and δ2H in CH4 at Imprint Analytics GmbH (Neutal, Austria) using a gas chromatograph (Shimadzu GC 2010Plus, Shimadzu Corp., Kyoto, Japan) coupled to a stable isotope ratio mass spectrometer (Nu Horizon; Nu Instruments Limited, Wrexham, UK). Gas samples were directly injected by an AOC5000 Autosampler (CTC Analytics, Zwingen, Switzerland) from the Labco Exetainers into the S/Sl Injector of the GC. The CH4 was separated from other gases in a Q-Plot GC column (Supelco, Bellefonte USA) at 35°C (isothermal). The separated analytes were identified in a quadrupole mass spectrometer (Shimadzu GCMS-QP2010Ultra). Methane was oxidized to CO2 for the δ13C analyses in an oxidation oven filled with oxidized Ni and Pt wires at a temperature of 1040°C. For the δ2H analysis, CH4 was pyrolysed at 1400°C to H2 in a ceramic tube (Hekatech, Wegberg, Germany). As reference material hexane vapour with a known δ13C and δ2H composition was used. δ13C is reported in ‰ VPDB, δ2H in ‰ SMOW.
9
Quantitative Analysis of Gaseous and Liquid Products
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The gas products
(hydrogen, carbon monoxide, methane, ethylene) produced in the cathodic
compartment were detected by an online gas chromatograph (GC, Agilent
8860) equipped with a thermal conductivity detector (TCD) and a flame
ionization detector (FID). The liquid products were measured depending
on the electrolyte composition. For ligand-free electrolyte, formate
was analyzed with high-performance liquid chromatography (HPLC, Shimadzu
Prominence) and alcohols and aldehydes were quantified using a liquid
GC (Shimadzu 2010 Plus). For ligand-containing electrolyte, the liquid
products were analyzed by 1H NMR (Bruker, 600 MHz) with
the water suppression technique and DMSO as internal standard.
(hydrogen, carbon monoxide, methane, ethylene) produced in the cathodic
compartment were detected by an online gas chromatograph (GC, Agilent
8860) equipped with a thermal conductivity detector (TCD) and a flame
ionization detector (FID). The liquid products were measured depending
on the electrolyte composition. For ligand-free electrolyte, formate
was analyzed with high-performance liquid chromatography (HPLC, Shimadzu
Prominence) and alcohols and aldehydes were quantified using a liquid
GC (Shimadzu 2010 Plus). For ligand-containing electrolyte, the liquid
products were analyzed by 1H NMR (Bruker, 600 MHz) with
the water suppression technique and DMSO as internal standard.
10
Seasonal BTEX Monitoring in Arad City
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The present study determined BTEX concentrations (benzene, toluene, ethylbenzene, and xylene) in different seasons—winter, spring, summer, and autumn—in the main crossroad from Arad City, from January to December 2016. Every month, there were four sampling times (every time, four samples were collected simultaneously—in total, 16 samples). Sampling was carried out at the height of 2 m above ground level by using air pumps (SKC 1003, SKC Inc., Houston, TX, USA), and stainless steel tubes (10.5 cm length, 4 mm inner diameter, Supelco, Bellefonte, PA, USA) filled with a mixture 2:1:1 w/w/w, Carbotrap C: Carbopack C: Carbotrap X adsorbents (Supelco, Bellefonte, PA, USA), with a flow rate of 200 mL min−1 for 60 min. Before use, tubes were conditioned for 30 min at 350 °C in a pure He flow of 50 mL min−1. Four samples were collected every time, and the tubes were analyzed on the same day as the sampling. The separation and detection of the compounds were performed by GC–MS (Shimadzu 2010 plus, GCMSTQ8040, Tokyo, Japan) coupled with a thermal desorption system (Shimadzu TD20, Kyoto, Japan) as described in Kannaste et al. [35 ].
To establish correlations between BTEX and the formation of tropospheric ozone in different seasons, the ozone concentration data were taken from open access data available athttp://www.anpm.ro (accessed date: 17 June 2017).
To establish correlations between BTEX and the formation of tropospheric ozone in different seasons, the ozone concentration data were taken from open access data available at
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