Cells were plated in fresh media and incubated for 48 h. Protein quantification was performed using proxy wells for normalization. Norvaline was used as an internal standard. Citrate and lactate standards were used to generate a standard curve for quantification. Metabolite extraction was performed using HPLC-grade ethanol (Sigma Aldrich). Samples were derivatized with methoxamine (PI45950, Thermo Fisher Scientific) and N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide with 1% tert-butyldimethylchlorosilane (Sigma Aldrich). Samples were analyzed by GC/MS using a HP-5MS Ultra Inert GC column (19091S-433UI, Agilent Technologies) installed in an Agilent 7890B gas chromatograph coupled to an Agilent 5977B mass spectrometer. Helium was used as the carrier gas. One microliter of sample was injected at 280°C. After injection, the GC oven was held at 60°C for 1 min. The oven ramped up to 320°C at 10°C/min and held for 9 min. The MS system operated under electron impact ionization mode at 70 eV, and the MS source and quadrupole were held at 230°C and 150°C, respectively. Peak abundance was determined by automated integration using MassHunter software (Agilent). Total abundance was normalized to the norvaline internal standard. Secretion rate was calculated by taking into account the specific growth rate, as determined by pre- and post-assay protein quantification.
Hp 5ms ultra inert gc column
Manufactured by Agilent Technologies
The HP-5MS Ultra Inert GC column is a capillary column designed for gas chromatography applications. It is characterized by its ultra-inert surface, which helps to minimize analyte adsorption and ensure reliable and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
Hplc grade ethanol, by Merck Group (2 mentions)
Agilent 7890b gas chromatograph, by Agilent Technologies (2 mentions)
Methoxamine, by Thermo Fisher Scientific (2 mentions)
Agilent 5977b mass spectrometer, by Agilent Technologies (2 mentions)
Pi45950, by Thermo Fisher Scientific (2 mentions)
Masshunter software, by Agilent Technologies (2 mentions)
Trace gc ultra, by Thermo Fisher Scientific (1 mentions)
Isq single quadruple ms, by Thermo Fisher Scientific (1 mentions)
5 protocols using hp 5ms ultra inert gc column
1
Targeted Metabolite Profiling by GC-MS
2
Quantifying Environmental Pollutants by GC-MS
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DDT,
DEHP, or PCBs were dissolved in methanol at 10,000 ppm and diluted
with ultrapure water to 1, 10, and 100 ppm. To these chemical pollutant
solutions (10 mL), CaO was added at 2 g L–1 and
vortexed for 1 min. Sodium polyphosphate was added at 2 g L–1 to the mixed solutions before hand-shaking and vortexing for 1 min.
The mixtures were left to stand at room temperature for 10 min and
filtered through qualitative filter paper.
For gas chromatography–mass
spectrometry (GC–MS), the chemical pollutants contained in
2 mL of the filtrates were extracted using 2 mL of hexane with hand-shaking
and vortexing. The extracted solutions were analyzed using a 7890B
GC–5977A MS system equipped with an HP-5 ms Ultra Inert GC
column (Agilent Technologies, CA), using helium as the carrier gas
and an electron ionization ion source. The oven temperature was controlled
as follows: 100 °C for 1 min, from 100 to 160 °C at 30 °C
min–1, from 160 to 270 °C at 5 °C min–1, and 270 °C for 3 min. For PCBs, single ion
monitoring was performed at 292, 326, and 360 m/z, which correspond to tetrachlorobiphenyls, pentachlorobiphenyls,
and hexachlorobiphenyls, respectively. Electron ionization mass spectra
were analyzed using Agilent MassHunter Qualitative Analysis software
for chemical species identification. The concentrations of the chemical
pollutants were quantified by using calibration curves.
DEHP, or PCBs were dissolved in methanol at 10,000 ppm and diluted
with ultrapure water to 1, 10, and 100 ppm. To these chemical pollutant
solutions (10 mL), CaO was added at 2 g L–1 and
vortexed for 1 min. Sodium polyphosphate was added at 2 g L–1 to the mixed solutions before hand-shaking and vortexing for 1 min.
The mixtures were left to stand at room temperature for 10 min and
filtered through qualitative filter paper.
For gas chromatography–mass
spectrometry (GC–MS), the chemical pollutants contained in
2 mL of the filtrates were extracted using 2 mL of hexane with hand-shaking
and vortexing. The extracted solutions were analyzed using a 7890B
GC–5977A MS system equipped with an HP-5 ms Ultra Inert GC
column (Agilent Technologies, CA), using helium as the carrier gas
and an electron ionization ion source. The oven temperature was controlled
as follows: 100 °C for 1 min, from 100 to 160 °C at 30 °C
min–1, from 160 to 270 °C at 5 °C min–1, and 270 °C for 3 min. For PCBs, single ion
monitoring was performed at 292, 326, and 360 m/z, which correspond to tetrachlorobiphenyls, pentachlorobiphenyls,
and hexachlorobiphenyls, respectively. Electron ionization mass spectra
were analyzed using Agilent MassHunter Qualitative Analysis software
for chemical species identification. The concentrations of the chemical
pollutants were quantified by using calibration curves.
3
Targeted Metabolite Profiling by GC-MS
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Cells were plated in fresh media and incubated for 48 h. Protein quantification was performed using proxy wells for normalization. Norvaline was used as an internal standard. Citrate and lactate standards were used to generate a standard curve for quantification. Metabolite extraction was performed using HPLC-grade ethanol (Sigma Aldrich). Samples were derivatized with methoxamine (PI45950, Thermo Fisher Scientific) and N-tert-butyldimethylsilyl-N-methyltrifluoroacetamide with 1% tert-butyldimethylchlorosilane (Sigma Aldrich). Samples were analyzed by GC/MS using a HP-5MS Ultra Inert GC column (19091S-433UI, Agilent Technologies) installed in an Agilent 7890B gas chromatograph coupled to an Agilent 5977B mass spectrometer. Helium was used as the carrier gas. One microliter of sample was injected at 280°C. After injection, the GC oven was held at 60°C for 1 min. The oven ramped up to 320°C at 10°C/min and held for 9 min. The MS system operated under electron impact ionization mode at 70 eV, and the MS source and quadrupole were held at 230°C and 150°C, respectively. Peak abundance was determined by automated integration using MassHunter software (Agilent). Total abundance was normalized to the norvaline internal standard. Secretion rate was calculated by taking into account the specific growth rate, as determined by pre- and post-assay protein quantification.
4
GC-MS Analysis of Solidified Multiflower Extract
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The GC-MS analysis of SME was performed with a Thermo Scientific, Trace GC Ultra & ISQ Single Quadruple MS. Helium (99.999%) was used as the carrier gas with a flow rate at 1.7 ml min−1.1 μl injected (1:10) using the splitless injection technique; Agilent 19091S-433UI column (HP-5ms Ultra Inert GC Column, 30 m, 0.25 mm, 0.25 µm, 7-inch cage). Temperatures: injector: 250°C, detector: 280°C, column: 100°C with a temperature gradient of 20°C.min−1, 260°C held for 10 min. The total GC running time is at 20 min. The MS ionization voltage was 70 eV. The MS scan parameters included a mass range of m/z 40–1000 atomic mass units (amu). Identification of compounds was matched using the database of Wiley9, replib, and mainlib Libraries. The name, retention time, and area under peak of the components of the test materials were ascertained.
5
Crude Oil Biodegradation Analysis by GC-MS
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Oil fractionation results were analyzed using an Agilent GC-MS System 7890A with an Agilent Inert MSD 5975C detector, HP-5MS Ultra Inert GC column (60 m × 0.250 mm × 0.25 μm), and ultra-high pure (UHP) helium as the carrier gas. This method defined the composition and types of compounds contained in the sample before and after biodegradation. The gas chromatography oven was programmed at 16.08 kPa with a column flow of 1 mL min−1. The temperature began at 40 °C and rose to 250 °C; the samples were run at a rate of 10 °C min−1 and then held for 25 min. The temperature was increased again from 250 °C to 290 °C at a rate of 10 °C min−1 and then held for 30 min. The sample of oil used for GC-MS analysis was taken from the station with the highest percentage (Station 18544) of oil biodegradation. The unit of abundance in the results refers to total hydrocarbons.
Further analysis of chromatogram was carried out by calculating the abundance of pristane (Pr) and phytane (Ph), which are commonly used as markers to observe the biodegradation process in crude oil. The comparison of n-C17/Pr and n-C18/Ph abundance was carried out to ensure that the degradation process occurred biologically, and the carbon preference index (CPI) was analyzed to show the ability of bacteria to degrade odd-numbered and even-numbered alkanes at the same ratio.
Further analysis of chromatogram was carried out by calculating the abundance of pristane (Pr) and phytane (Ph), which are commonly used as markers to observe the biodegradation process in crude oil. The comparison of n-C17/Pr and n-C18/Ph abundance was carried out to ensure that the degradation process occurred biologically, and the carbon preference index (CPI) was analyzed to show the ability of bacteria to degrade odd-numbered and even-numbered alkanes at the same ratio.
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