The GC-MS analysis was performed using Shimadzu 2010 Plus gas chromatograph coupled to a Shimadzu QP2010 Ultra mass selective detector. The gas chromatography-flame ionization detector (GC-FID) was used. The fibre containing the extracted aroma compounds was injected into GC/MS injector. The split mode was used. The separation was performed by means of a Restek Rxi-5MS capillary column, 60 m length, 0.25 mm i.d., and a 0.25 μm phase thickness. The oven program was as follows: Initial temperature was 60 °C for 2 min, which was increased to 240 °C at 3 °C min -1 . 250 °C was maintained for 4 min. Helium (99.999%) was used as carrier gas with a constant flow-rate of 1 μL min -1 . Detection was carried out in electronic impact mode (EI) and ionization voltage was fixed to 70 eV. Scan mode (40-450 m/z) was used for mass acquisition. The volatile compounds were identified by comparison with the mass spectra of the two libraries (FFNSC1.2 and W9N11) and using the standard compounds (βpinene, camphene, limonene oxide, isopinocarveol, 1-terpinen-4-ol, β-ionone, caryophyllene oxide, biformene, and nonanal) (Renda et al., 2019) (link).
Rxi 5ms capillary column
Manufactured by Restek
Sourced in United States, Japan, Germany
The Rxi-5MS capillary column is a general-purpose fused silica column designed for a wide range of gas chromatography applications. It features a 5% diphenyl and 95% dimethyl polysiloxane stationary phase, providing a balance of polarity and thermal stability.
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Lab products found in correlation
Pegasus ht, by Leco (8 mentions)
Class 5000 chromatography workstation software, by Shimadzu (5 mentions)
Agilent 7890b gas chromatography, by Gerstel (5 mentions)
Qp2010 ultra, by Shimadzu (3 mentions)
Qp2010, by Shimadzu (3 mentions)
Gcms qp2010 ultra system, by Shimadzu (3 mentions)
Qp2010 ultra gc fid ms, by Shimadzu (2 mentions)
Agilent 7890b gas chromatograph, by Agilent Technologies (2 mentions)
Gc 2010 plus, by Shimadzu (2 mentions)
Aoc 5000 plus autosampler, by Shimadzu (2 mentions)
Gc 2010plus gcms qp2010 ultra system, by Shimadzu (2 mentions)
Multipurpose sample mps2 with dual heads, by Gerstel (2 mentions)
Optima lc ms, by Thermo Fisher Scientific (2 mentions)
Repeater xstream electronic pipette, by Eppendorf (2 mentions)
Multipurpose sample mps2, by Gerstel (2 mentions)
Aoc 5000, by Shimadzu (2 mentions)
Qp2010 ultra mass, by Shimadzu (1 mentions)
2010 plus, by Shimadzu (1 mentions)
Trace 1300 gas chromatograph, by Thermo Fisher Scientific (1 mentions)
Isq lt mass spectrometer, by Thermo Fisher Scientific (1 mentions)
42 protocols using rxi 5ms capillary column
1
GC-MS Analysis of Aroma Compounds
2
Characterization of Culture Media Metabolites
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The culture media supernatant was analyzed by GC-MS [29 ] and ionic exchange chromatography [30 (link)] to assess any degradation metabolites. For GC-MS, the supernatant was extracted with ethyl acetate at a 1 : 2 ratio, stirred every 15 min for 8 h, and left still for 24 h. The organic phase was separated and injected (1 μL) on a Trace 1300 gas chromatograph (Thermo Scientific, USA) coupled to an ISQ-LT mass spectrometer (Thermo Scientific, USA). The extracted components were separated on a Rxi-5 ms capillary column (60 m × 0.25 mm ID × 0.25 μm film, Restek, USA) based on temperature programming, while the initial temperature of 40°C was kept constant for 5 min, increased to 200°C at a constant rate of 10 °C/min, followed by 260°C at a constant rate of 5°C/min, and finally to 300°C at a constant rate of 10°C/min. The mass analyzer consisted of a single quadrupole, using electron impact ionization at 70 eV. For ionic exchange chromatography, the supernatants were injected into an 883 Basic IC Plus apparatus (Metrohm AG, CH) with a MetroSep A Supp 5–150/4.0 column (Metrohm AG, CH) and analyzed using the anionic analysis method with the MagIC Net 3.3 software (version 3.3, Metrohm AG, CH).
3
GC-MS Analysis of AO7 Degradation
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The products of AO7 degradation by NYC-1consortium were analyzed by gas chromatography/mass spectrometry (GC–MS). The samples were extracted with dichloromethane (DCM) using liquid to liquid extraction at various pH values to neutralize any charged compounds, such as phenolics and amines. The extracted metabolites were analyzed using a gas chromatograph system (Agilent) equipped with a Restek Rxi-5 ms capillary column. A sample volume (1 μL) was injected in split mode with injector temperature of 250 °C. The flow rate of helium used as a carrier gas was 1.1 mL/min. The initial temperature was held at 40 °C for 1 min, then ramped at 10 °C per min until 340 °C (hold time 10 min). Electron ionization (EI) was used with MS source temperature at 230 °C and Quad temperature at 150 °C. The metabolites were identified by comparison of the obtained spectra with those stored in the NIST mass spectra database.
4
Glycosidic Linkage Determination by GC-MS
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Glycosidic linkage determination was performed through methylation studies. The fractions most active in the complement assay from each extract were selected for linkage determination. Prior to methylation, the uronic acids were reduced with NaBD 4 to their corresponding neutral sugars, dideuterated in position 6. After reduction of the polymers, methylation, hydrolysis, reduction using sodium borodeuteride and acetylation were carried out [13] . The derivatives were analyzed by GC-MS using a GCMS-QP2010 (Shimadzu) in which was attached a Restek Rxi-5MS capillary column (30 m; 0.25 mm i.d.; 0.25 μm film). The injector temperature was 280 °C, the ion source temperature 200 °C and the interface temperature 300 °C. The column temperature was 80 °C when injected, then increased at 10 °C/min to 140 °C, followed by 4 °C/min to 210 °C, and then 20 °C/min to 300 °C. Helium was the carrier gas (pressure control: 80 kPa.) The compounds represented by each peak were characterized by their retention times and interpretation of their characteristic mass spectra. The estimation of the relative amounts of each linkage type was related to the total amount of each monosaccharide type as determined by methanolysis. Effective carbon-response factors were applied for quantification [23] .
http://repository.uwc.ac.za
http://repository.uwc.ac.za
5
GC-MS Analysis of Clove Essential Oil
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Gas chromatography-mass spectrometry (GC-MS) analysis of Eugenia caryophyllata essential oil was carried out using a Shimadzu 2010 Plus gas chromatography coupled to a Shimadzu QP2010 Ultra mass selective detector. The separation was performed by means of a Restek Rxi-5MS capillary column, 60 m length, 0.25 mm i.d. and a 0.25 μm phase thickness. The split mode was used. The oven was programmed as follows: Initial temperature was 60 o C for 2 min, which was increased to 240 o C at 3 o C min -1 , 250 o C was maintained for 4 min. Helium (99.999%) was used as carrier gas with a constant flow-rate of 1 mL min -1 . Detection was carried out in electronic impact mode (EI); ionization voltage was fixed to 70 eV. Scan mode (40-450 m/z) was used for mass acquisition. The volatile compounds were identified by comparison of their retention indices (relative to C7-C30 alkane standards), and matching mass spectral data with those held in FFNSC1.2 and W9N11 library of mass spectra and literature comparison [8] . This part of the study has been performed in the department of chemistry, Faculty of Sciences, Karadeniz Technical University, Trabzon Turkey.
6
GC-FID/MS Analysis of Essential Oils
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EO analysis was carried out using a Shimadzu QP2010 ultra GC-FID/MS, Shimadzu 2010 plus FID fitted with a PAL AOC-5000 plus auto sampler, and Shimadzu Class-5000 Chromatography Workstation software (Shimadzu Corp., Kyoto, Japan). The separation was analyzed by means of a Restek Rxi-5MS capillary column (30 mm × 0.25 mm × 0.25 μm) (Restek Corp., Bellefonte, PA, USA). The essential oil injections to GC-FID/MS wereperformed in split mode (1:30) at 230 °C. The essential oil solution (1 μL) in
n-hexane (HPLC grade) was injected and analyzed with the column held initially at 60 °C for 2 min and then increased to 240 °C with a 3 °C/min heating ramp. The oven program was as follows: the initial temperature was 60 °C for 2 min, which was increased to 240 °C for3 min; the final temperature of 250 °C was held for 4 min. Helium (99.999%) was used as the carrier gas, with a constant flowrate of 1 mL/min. Detection was implemented in electronic impact mode (EI); ionization voltage was fixed at 70 eV and scan mode (40–450
m/z) was used for mass acquisition. Each sample was analyzed and the mean of each reported.
n-hexane (HPLC grade) was injected and analyzed with the column held initially at 60 °C for 2 min and then increased to 240 °C with a 3 °C/min heating ramp. The oven program was as follows: the initial temperature was 60 °C for 2 min, which was increased to 240 °C for3 min; the final temperature of 250 °C was held for 4 min. Helium (99.999%) was used as the carrier gas, with a constant flowrate of 1 mL/min. Detection was implemented in electronic impact mode (EI); ionization voltage was fixed at 70 eV and scan mode (40–450
m/z) was used for mass acquisition. Each sample was analyzed and the mean of each reported.
7
Fecal Metabolomic Profiling of Pediatric Participants
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Fresh fecal samples from each participant (37 PP and 25 control girls) were collected for untargeted metabolomic analysis by Metabo-Profile. Briefly, an Agilent 7890B gas chromatograph and a time-of-flight mass spectrometry system (GC-TOF/MS, Pegasus HT, Leco Corp., USA) were used for untargeted metabolomic analysis. The chromatographic column was an Rxi-5ms capillary column (inner diameter 30 m × 250 μm, film thickness 0.25 μm; Restek Corporation, USA). Helium was maintained at a constant flow rate of 1.0 mL/min. The temperature for both injection and transfer interface was set to 270°C. Measurements were performed in full scan mode (m/z 50–500) using electron impact ionization (70 eV). Quality control procedures were performed through sample preparation and analytical testing. The raw data generated by GC-TOF/MS were processed for metabolite characterization and quantification using the XploreMET software (Metabo-Profile, Shanghai, China). Statistical analysis was performed using Student’s t-test to identify DEMs, and statistical differences between the two groups were defined as p < 0.05 and |log2FoldChange| > 1.
8
GC-FID Analysis of Extracted Aroma Compounds
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The gas chromatography chromatography-flame ionization detector (GC-FID) analysis was carried out on a Shimadzu QP2010 plus gas chromatography coupled to a Shimadzu QP2010 ultra mass selective detector. The fiber-containing, extracted aroma compounds were injected into the GC-MS injector (split mode). Separation took place with a Restek Rxi-5MS capillary column of 30 m length, 0.25 mm i.d., and 0.25 μm phase thickness. The oven program began at an initial temperature of 60 °C for 2 min, which was then increased to 240 °C for three minutes, and then a final temperature of 250 °C for four minutes. The injector temperature was 280 °C and the split ratio was 1:20. The carrier gas was helium (99.999%), with a constant flow rate of 1 mL/min; sample size was 1 μL. Detection was processed in electronic impact mode (EI); ionization voltage was fixed to 70 eV, and scan mode (40–450 m/z) was used for mass acquisition.
9
GC-MS Analysis of Derivatized Carbohydrates
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The OS samples were further analyzed using GC-MS, a gas chromatograph (Agilent 7890A) coupled with the mass selective (Agilent 5975C MSD) detector. The compounds were separated on Rxi5-MS capillary column (30 m × 0.25 mm, 0.25 µm, Restek). Helium was used as a carrier gas at a 1.3 mL/min flow rate. Before the analysis, the samples were derivatized with trimethylsilylation reagent, which consisted of pyridine, BSA, and TMCS (20:20:1). Analytical equipment was controlled, and data were analyzed using MassHunter Acquisition and Data Analysis software (MassHunter 12.0, Agilent, Santa Clara, CA, USA). The GC-MS was used for monitoring the purified fractions for the presence of mono- and disaccharides. GC-MS cannot be and thus was not used to quantitative determine HMOs, even after chemical derivatization, due to their high molecular weights.
10
GC-TOFMS Analysis of Organic Compounds
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The separation analysis was performed using an Agilent 7890N gas chromatograph in splitless mode connected to a time-of-flight mass spectrometer (LECO Corp., St. Joseph, MI). Separation was achieved using an Rxi-5ms capillary column (Crossbond® 5% diphenyl/95% dimethyl polysiloxane; Restek), with helium as the carrier gas at a constant flow rate of 1.0 mL/min. The GC oven temperature program was as follows: 2 min at 80 °C, raised by 10 °C/min to 140 °C, by 4 °C/min to 180 °C, by 10 °C/min to 240 °C and by 25 °C/min to 290 °C, followed by hold for 4.5 min at 290 °C. The temperatures of the injection, transfer interface, and ion source were set to 270, 270, and 220 °C, respectively. Mass spectra were obtained with electron impact ionization (70 eV) at full scan mode (m/z 30–600) and an acquisition rate of 25 spectra/s.
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