Confluent cells in 6-well plated were homogenized in 0.5 mL of chilled 80% (v/v) methanol. The samples were centrifuged at 12,000 rpm for 10 min and the supernatants were transferred to a high recovery glass sampling vial (CNW, VAAP-31509-1232-100) to vacuum dry at room temperature. The residue was re-suspended with 30 μL pyridine containing 20 mg/mL methoxyamine hydrochloride (Sigma-Aldrich, 226904) at 37 °C overnight and further derivatized with 20 μL of N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide (Sigma-Aldrich, Cat#: 394882) at 70 °C for 30 min. Then 1 μL aliquot of the derivatized sample was injected into Agilent 7890 A gas chromatography coupled with Agilent 5975 C mass spectrometer. Separation was achieved on a HP-5ms fused-silica capillary column with the helium as the carrier gas at a constant flow rate of 1 mL/min through the column.
Agilent 5975c mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 5975C mass spectrometer is a high-performance instrument designed for analytical laboratories. It provides accurate mass analysis and identification of chemical compounds. The 5975C utilizes electron ionization (EI) and chemical ionization (CI) techniques to generate and detect ions. It features a quadrupole mass analyzer and an electron multiplier detector. The instrument is capable of performing full-scan, selected ion monitoring, and tandem mass spectrometry (MS/MS) analyses.
Automatically generated - may contain errors
Lab products found in correlation
Agilent 7890a, by Agilent Technologies (3 mentions)
Agilent 7890a gas chromatograph, by Agilent Technologies (3 mentions)
Hp 5ms gas chromatographycolumn, by Agilent Technologies (2 mentions)
Agilent db 35ms column, by Agilent Technologies (2 mentions)
Mox reagent, by Thermo Fisher Scientific (2 mentions)
Tert butyldimethylchlorosilane, by Merck Group (2 mentions)
Db 35ms column, by Agilent Technologies (2 mentions)
Mps autosampler, by Agilent Technologies (2 mentions)
Hp 5ms capillary column, by Agilent Technologies (2 mentions)
N tert butyldimethylsilyl n methyltrifluoroacetamide, by Merck Group (1 mentions)
Methoxyamine hydrochloride, by Merck Group (1 mentions)
Agilent 7890a gas chromatography, by Agilent Technologies (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Methoxyamine hydrochloride, by Thermo Fisher Scientific (1 mentions)
Msk a2 1, by Cambridge Isotopes (1 mentions)
7890a gas chromatograph, by Agilent Technologies (1 mentions)
Agilent 1100 series, by Agilent Technologies (1 mentions)
7890a gas chromatograph gc, by Agilent Technologies (1 mentions)
Isq lt, by Thermo Fisher Scientific (1 mentions)
Xad 2 tubes, by SKC (1 mentions)
31 protocols using agilent 5975c mass spectrometer
1
GC-MS Analysis of Metabolites in Cultured Cells
2
Synthesis and Characterization of Novel Benzothiazole Derivatives
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All reagents and organic solvents were purchased from Sigma Chemical Co. (St. Louis, USA) and used without further purification. Thin-layer chromatography (TLC) was carried out on pre-coated silica gel aluminum plates (Merck silica gel 60, F254). Melting points of target compound 8a–o were measured on a Kofler hot stage apparatus and were uncorrected. 1H NMR and 13C NMR spectra were recorded on Bruker FT-500 spectrometer (Bruker, Rheinstetten, Germany) in DMSO-d6 with tetramethylsilane (TMS) as the internal standard. IR spectra were recorded on Nicolet Magna FTIR 550 spectrophotometer (resolution 2 cm−1) in KBr pellets. Elemental analysis was carried out with an Elemental Analyzer system GmbH VarioEL CHNS mode (Germany). Mass spectra were recorded on Agilent 5975C Mass Spectrometer.
3
Methanol-based Metabolite Extraction and GC-MS Analysis
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Frozen tissue samples (20–30 mg) were homogenized for 2 min using ceramic beads (Precellys 2 mL Hard Tissue Homogenizing Ceramic Beads Kit, Bertin Instruments, US) in 500 μL −20 °C methanol, ice-cold 400 μL saline and ice-cold dH2O containing amino acid isotope labelled internal standards (Cambridge Isotope Laboratories, #MSK-A2-1.2). An aliquot of homogenate (50 μL) was dried under air and resuspended in RIPA buffer for protein quantification using BCA assay (BCA Protein Assay, Lambda, Biotech Inc., US). To the remaining homogenate, 1 mL of chloroform was added and the samples were vortexed for 5 min followed by centrifugation at 4 °C for 5 min @ 15 000g. The organic phase was collected and the remaining polar phase was re-extracted with 1 mL of chloroform. An aliquot of the polar phase was collected and vacuum-dried at 4 °C and subsequently derivatized with 2% (w/v) methoxyamine hydrochloride (Thermo Scientific) in pyridine for 60 min following by 30 min sialyation N-tertbutyldimethylsilyl-N-methyltrifluoroacetamide (MTBSTFA) with 1% tert-butyldimethylchlorosilane (tBDMS) (Regis Technologies) at 37 °C. Polar derivatives were analyzed by GC–MS using a DB-35MS column (30m × 0.25 mm i.d. X 0.25 μm, Agilent J&W Scientific) installed in an Agilent 7890A gas chromatograph(GC) interfaced with an Agilent 5975C mass spectrometer (MS) as previously described [24 (link)].
4
Monosaccharide Composition Analysis of MLM and MFM
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Monosaccharide composition of MLM and MFM was analyzed using a modified GC-MS analytical procedure previously adopted by Xia et al. [73 (link)] depending upon trimethylsilyl dithioacetal (TMSD) derivatization. A 1 µL volume of the sample was applied to Agilent 7890A Gas Chromatography coupled to Agilent 5975C Mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) with HP-5MS column using a temperature range of 80 °C for 0 min, 80–90 °C at 2.5 °C/min, 190–252 °C at 2 °C/min, 252–300 °C at 25 °C/min, 300–310 °C at 25 °C/min and held for 15 min. Mass spectra were recorded employing total ion chromatogram (TIC) mode and interpreted using NIST 5 software [25 (link)].
5
Quantifying Trehalose Uptake by GC-MS
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Trehalose uptake quantification by gas chromatography and mass spectrometry was performed precisely as reported19 (link). 13C12 trehalose (Omicron Biochemical, South Bend IN) was used as a sample extraction and derivatization internal standard. 5 nmol internal standard were added to the samples which were then extracted into 200 μl isopropanol:CH3CN:H2O. Samples were centrifuged and the supernatant was taken to dryness under nitrogen. Samples were derivatized using 100 ul N-Methyl-N-(Trimethysilyl) trifluoroacetamide (MSTFA): 10% pyridine in CH3CN (1:3) at room temperature overnight. Derivatized samples were analyzed using an Agilent 7890 A gas chromatograph interfaced to an Agilent 5975 C mass spectrometer. The GC column was a HP-5MS (30 m, 0.25 mm internal diameter, 0.25 um film coating, P.J. Cobert, St. Louis, MO). A linear temperature gradient was used. The initial temperature (80 °C) was held for 2 min and increased to 300 °C at 10 °C/min. The temperature was held at 300 °C for 2 min. Samples were run by electron ionization (EI) mode and the source temperature, electron energy and emission current were 200 °C, 70 eV and 300 μA, respectively. The injector and transfer line temperatures were 250 °C. The ions monitored were m/z 361 and 13-Trehalose at m/z 367.
6
Quantifying L-Amino Acids in Mouse Tissues by GC-MS
7
Deuterium Labeling of Water via Acetone Exchange
8
Profiling Acylcarnitines and Organic Acids
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Blood and urine samples were collected to measure the levels of 26 Acylcarnitines (ACs) and 76 urine organic acids (UOA). Whole blood and serum were collected in serum tubes and stored between 2 and 8 °C. Fresh urine samples were collected in a test tube and stored frozen. All reagents and internal standards used for LC–MS/MS and for GC–MS were as previously described [15 (link)].
The ACs, extracted and derivatized to butyl esters as previously described [15 (link), 16 (link)], were analyzed on an API 4000 triple quadrupole mass spectrometer (Applied Biosystems-Sciex, Toronto, Canada) coupled with the high performance liquid chromatograph Agilent 1100 series (Agilent Technologies, Waldbronn, Germany).
Creatinine concentration of urine samples was determined by using automatic analysis system BM/Hitachi 904. Organic acids were extracted from urine and analysed on a GC–MS system including an Agilent 7890A (Agilent Technologies, Santa Clara, CA, USA) gas chromatograph and an Agilent 5975C mass spectrometer. Extraction and analysis were performed as previously described [17 ]; the concentration of organic acids was normalized to the creatinine concentration of the urine sample and expressed as mmol organic acid/mol creatinine.
The ACs, extracted and derivatized to butyl esters as previously described [15 (link), 16 (link)], were analyzed on an API 4000 triple quadrupole mass spectrometer (Applied Biosystems-Sciex, Toronto, Canada) coupled with the high performance liquid chromatograph Agilent 1100 series (Agilent Technologies, Waldbronn, Germany).
Creatinine concentration of urine samples was determined by using automatic analysis system BM/Hitachi 904. Organic acids were extracted from urine and analysed on a GC–MS system including an Agilent 7890A (Agilent Technologies, Santa Clara, CA, USA) gas chromatograph and an Agilent 5975C mass spectrometer. Extraction and analysis were performed as previously described [17 ]; the concentration of organic acids was normalized to the creatinine concentration of the urine sample and expressed as mmol organic acid/mol creatinine.
9
GCMS Analysis of Polar Metabolites
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Polar metabolites were analyzed by gas chromatography mass spectrometry (GCMS). Dried extracts were derivatized with 16 μL MOX reagent (Thermo) for 60 minutes at 37°C followed by 20 μL N-tert-Butyldimethylsilyl-N-methyltrifluoroacetamide with 1% tert-Butyldimethylchlorosilane (Sigma) for 30 minutes at 60 °C. Following derivatization, samples were analyzed using an Agilent 7890A gas chromatograph using a DB-35MS column (Agilent) coupled to an Agilent 5975C mass spectrometer. Mass isotopomer distributions and total ion counts were determined by integrating metabolite ion fragments and were corrected for natural abundance.
10
PAH and BTEX Exposure Analysis
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Air samples for PAH analysis were collected during the entire duration of the exposure (180 min) on Teflo air sampling membranes (PTFE, 37 mm diameter, 2 µm pore size, Pall Corp., Port Washington, NY, USA) followed by XAD-2 tubes (SKC Inc., Dorset, UK). Particle filters and XAD tubes were extracted using dichloromethane analyzed for 32 native PAHs (including the 16 U.S. EPA priority PAHs) and 16 alkylated species. Target compounds were separated on an Agilent 5975C mass spectrometer (MS) coupled to a 7890A gas chromatograph (GC, Agilent Technologies, Santa Clara, CA, USA) [11 ]. Analyses of benzene, toluene, ethyl benzene, m + p xylene, and o-xylene (BTEX) were performed by the Swedish Environmental Institute (IVL). The samples were collected on Tenax® TA thermal desorption tubes (SKC Inc., Dorset, UK) and analyzed by thermal desorption GC-MS. Target compounds were separated on a non-polar analytical mass spectrometer (TraceGold, TG-1MS, Thermo Fisher Scientific Inc., Franklin, MA, USA) coupled to an ISQ LT (Thermo Fisher Scientific Inc., Franklin, MA, USA) [11 ].
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