Metabolomics profiles of 18 free amino acids (FAAs), 16 free fatty acids (FFAs), and 5 carbohydrates in the 3rd instar larvae of O. furnacalis in response to PDS treatment were detected using targeted GC–MS techniques. Dried larvae were ground, and samples were incubated in trimethylsilane solutions [pyridine:hexamethyldisilazane:trimethylchlorosilane = 9:3:1 (v/v/v)] at 100°C for 30 min. Samples were then derivatized under nitrogen and dissolved into dichloromethane for analysis. GC–MS analysis was performed using a DB-5MS GC column (30 m × 0.25 mm, 0.25-μm film thickness; J&W Scientific, Folsom, CA, USA) with a temperature increase from 80°C to 280°C and a flow rate of 1.0 ml/min. The carrier gas was helium (He). GC–MS analysis was performed using a Thermo Scientific Trace GC ultra ISQ mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) equipped with a quadrupole mass selective detector on electro-impact mode (70 eV). Based on these analyses, metabolic compounds were identified by comparing them to the standards in the national Mass Spectral Librarie (NIST 2008 database: www.sisweb.com/software/ms/nist.htm ). The percentages were calculated using the peak area normalization method. Each experiment was replicated 5 times.
Db5 ms gc column
Manufactured by Agilent Technologies
Sourced in United States
The DB5-MS GC column is a capillary column designed for gas chromatography (GC) applications. It features a 5% phenyl, 95% dimethylpolysiloxane stationary phase, which provides a balance of polarity and inertness for a wide range of analytes. The column offers high thermal stability and low bleed, making it suitable for a variety of GC analyses.
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Lab products found in correlation
7890 gc system, by Agilent Technologies (2 mentions)
5977 ms system, by Agilent Technologies (2 mentions)
Trace gc ultra isq mass spectrometer, by Thermo Fisher Scientific (1 mentions)
5975c msd, by Agilent Technologies (1 mentions)
Agilent 7890a gc, by Agilent Technologies (1 mentions)
5975c mass spectrometer, by Agilent Technologies (1 mentions)
7890a gas chromatograph, by Agilent Technologies (1 mentions)
Agilent 7890 gc system, by Agilent Technologies (1 mentions)
Gc ms 7890a, by Agilent Technologies (1 mentions)
9 protocols using db5 ms gc column
1
Metabolomic Profiling of Insect Larvae under PDS Treatment
2
Phytochemical Profiling of L. fischeri Extracts
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The root and leaf extracts from L. fischeri were dissolved in hexane and subjected to GC-MS analysis using Agilent 7890A GC and 5975C MSD instruments (Agilent, Santa Clara, CA, USA). The nonpolar compounds were separated on a J&W DB-5ms GC column (60 m × 0.25 mm × 0.25 µm). Helium was used as the carrier gas, with a column-head pressure of 16.909 psi. The oven temperature was initiated at 50 °C for 5 min, programmed at 300 °C at a rate of 5 °C/min, maintained for 30 min, increased to 310 °C at a rate of 10 °C/min, and held for 10 min. The flow rate was set at 1 mL/min. A full scan mode (m/z 30–500) was used to detect all the target compounds. The compounds were tentatively detected by comparing the mass spectra of the peaks with those in the NIST library, and the percentage of each compound was calculated by normalizing the methods of at least three experiments.
3
GC-MS Analysis of Wristband Chemicals
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We quantitatively analyzed all wristband extracts for 94 organic chemicals using an Agilent (Santa Clara, CA, U.S.) 7890A gas chromatograph (GC) interfaced with an Agilent 5975C mass spectrometer (MS) detector as well as an Agilent 6890N GC interfaced with an Agilent 5975B MS. We used an Agilent DB‐5MS GC column (30 m × 0.25 mm), and each chemical in the method was calibrated with a curve of at least five points (correlations ≥0.99). The method includes 94 VOCs and SVOCs including PAHs, OPAHs, tri‐r‐phosphates, and alkanes. All target analytes, molecular weights, limits of detection (LODs), and limits of quantitation (LOQs) are in Table A2 of Supporting Information S1 .
In a few cases, we were not able to detect certain deuterated chemical surrogates in a minority of wristband extracts due to matrix interference. As a result, we were not able to quantify the target chemical related to the undetected chemical surrogate in these instances. Table A2 in Supporting InformationS1 includes the number of wristbands for each target chemical that did not have matrix interference. Compounds in sweat or personal care products are potential reasons for matrix interference.
In a few cases, we were not able to detect certain deuterated chemical surrogates in a minority of wristband extracts due to matrix interference. As a result, we were not able to quantify the target chemical related to the undetected chemical surrogate in these instances. Table A2 in Supporting Information
4
GC-MS Analysis of Cyclohexanone and Cyclohexanol
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A 500 μL
aliquot of the headspace gas from each headspace vial was injected
into the split/splitless inlet (20:1 split ratio) onto an Agilent
7890 GC system, fitted with a DB5-MS GC column 30 m × 0.25 mm,
0.25 μm (Agilent Technologies, Santa Clara, CA). The oven was
initially held at 40 °C for 1 min before being ramped from 10
°C for 1 min to 120 °C followed by further ramping from
40 to 250 °C where it was held for the remainder of the 15 min
run time. MS was operated in selected ion monitoring mode with an
electron impact source (70 eV) using m/z 55 and 98
for cyclohexanone and m/z 57 and 82 for cyclohexanol.
The source and quadrupole temperatures were 230 and 150 °C, respectively.Figure 5 shows an example
chromatogram.
aliquot of the headspace gas from each headspace vial was injected
into the split/splitless inlet (20:1 split ratio) onto an Agilent
7890 GC system, fitted with a DB5-MS GC column 30 m × 0.25 mm,
0.25 μm (Agilent Technologies, Santa Clara, CA). The oven was
initially held at 40 °C for 1 min before being ramped from 10
°C for 1 min to 120 °C followed by further ramping from
40 to 250 °C where it was held for the remainder of the 15 min
run time. MS was operated in selected ion monitoring mode with an
electron impact source (70 eV) using m/z 55 and 98
for cyclohexanone and m/z 57 and 82 for cyclohexanol.
The source and quadrupole temperatures were 230 and 150 °C, respectively.
chromatogram.
5
Bioactive Profiling of J. procera Leaf Extract
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Based on the results of the UV-spectrophotometer and ANOVA test, the ethanol seed extract was selected to be injected into GC-MS analysis for bioactive compound profiling by using gas chromatography-mass spectrometry (GC-MS 7890A; Agilent Technologies-USA, equipped with a 5975 mass-selective detector and a 7693 automated liquid sampler, fitted with a DB-5MS GC column (30 m length, 0.25 mm inner diameter, and 0.25 μm film thickness)). The extract was filtered using a 2 µm membrane filter. Then, a 1.0 µL aliquot of the ethanol extract was injected into the system. The injection temperature was 280 °C and the column temperature was adjusted to 300 °C. Helium gas was used as the carrier with a flow rate of 1 mL/min. The electron ionization energy was 70 eV while the GC-MS analysis leaf extract of J. procera was undertaken and published recently [43 (link)].
6
GC-MS Analysis of CBEE and CBAF
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An Agilent (7890B) gas chromatograph having an inert mass selective detector (5977B) and DB-5MS GC column, having a length of 30 m, internal diameter of 0.25 mm, and film thickness of 0.25 µm, was used for GC–MS analysis of CBEE and CBAF. A split less mode was used to inject 2 μL of each sample, while injector had the temperature of 250 °C and an interface temperature of 280 °C. In addition, the oven temperature was automated to start from 100 °C for 0.5 min, followed by a ramp of 20 °C per minute until it reached 340 °C. For electron spray ionization, helium carrier gas was incorporated at full-scan mode at –70 eV. The complete cycle for each sample was about 30 min.
7
Tracing Metabolism in Activated T Cells
8
GC-MS Analysis of Metabolite Profile
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An Agilent (7890B, Santa Clara, CA, USA) gas chromatograph equipped with an inert mass selective detector (5977B) and DB-5MS GC column (30 m in length, 0.25 mm in internal diameter, 0.25 µm in film thickness) was used for analysis. Sample injection (2 uL; methanolic fraction was prepared from fresh concentrate) was performed in the split-less mode, with an injector temperature of 250 °C and an interface temperature of 280 °C. The temperature of the oven was programmed from an initial value of 100 °C for 0.5 min andramped up to 340 °C at 20 °C/minute for 1 min. Helium was used as a carrier gas and electron impact ionization was used at −70 eV in full-scan mode. The run time was 30 min. Matching with National Institute of Standards and Technology (NIST) Library, Gaithersburg, MD 20899, USA, was carried out to identify the detected compounds.
9
GC-MS Metabolite Derivatization Protocol
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Dried metabolite extracts were resuspended in pyridine and derivatized with methoxyamine (sc-263468 Santa Cruz Bio) for 60 min at 37°C and subsequently with N-(tert-butyldimethylsilyl)-N-methyl-trifluoroacetamid, with 1% tert-utyldimethylchlorosilane (375934 Sigma-Aldrich) for 30 min at 80°C. Isotopomer distributions were measured using a DB5-MS GC column in a 7890 GC system (Agilent Technologies) combined with a 5977 MS system (Agilent Technologies). Data processing, including correction for natural isotope abundance was performed by an in-house R script.
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