Total FMO activity in liver homogenate quantified flavin-dependent d9-TMAO production from d9-TMA using tandem mass spectrometry (LC/MS/MS) analyses using d4-choline as internal standard, as described[12 (link)]. Stable isotope dilution LC/MS/MS was used to quantify TMA, TMAO and choline in plasma and animal diets, as described[12 (link),32 (link)]. Microbial choline TMA lyase activity was quantified by monitoring d9-TMA production from d9-choline substrate using LC/MS/MS analyses[15 (link)]. Reactions were stopped by addition of methanolic acidified d4-choline and 13C3,15N1-TMA internal standard mix, and deproteinated reaction mixtures analyzed on a Shimadzu (Columbia, MD) 8050 triple quadrupole mass spectrometer with UHPLC interface.
8050 triple quadrupole mass spectrometer
Manufactured by Shimadzu
Sourced in Japan, United States
The 8050 triple quadrupole mass spectrometer is an analytical instrument designed for sensitive and selective detection of target compounds. It features a triple quadrupole configuration to perform tandem mass spectrometry (MS/MS) analysis. The 8050 is capable of high-throughput quantitative analysis across a wide range of applications.
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Lab products found in correlation
Kinetex c8, by Phenomenex (3 mentions)
Nexera x2 lc 30ad, by Shimadzu (3 mentions)
D3 l carnitine, by Cambridge Isotopes (2 mentions)
Strata x cartridge, by Phenomenex (1 mentions)
Lcms 8050, by Shimadzu (1 mentions)
Hypersil gold c18, by Thermo Fisher Scientific (1 mentions)
Nexera ultra high performance liquid chromatograph, by Shimadzu (1 mentions)
Internal standards, by Cayman Chemical (1 mentions)
Nexera ultra high performance liquid chromatograph system, by Shimadzu (1 mentions)
Acquity uhplc beh c18 column, by Waters Corporation (1 mentions)
Nexera x2 uhplc, by Shimadzu (1 mentions)
Lc 20ad, by Shimadzu (1 mentions)
Lcms 8050 system, by Shimadzu (1 mentions)
28 protocols using 8050 triple quadrupole mass spectrometer
1
Quantifying Microbial TMAO Production
2
Quantification of TMA and TMAO by LC-MS/MS
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Stable isotope dilution high-performance LC-MS/MS was used for quantification of TMA and TMAO levels as previously described (3 (link), 44 (link)). Their d9(methyl)-isotopologues, d9-TMA and d9-TMAO, were spiked into plasma as internal standards. LC-MS/MS analyses were performed on a Shimadzu 8050 triple quadrupole mass spectrometer. TMA, TMAO, d9-TMA, and d9-TMAO were monitored using multiple reaction monitoring of precursor and characteristic product ions as follows: m/z 60.2 → 44.2 for TMA; m/z 69.0 → 49.1 for d9-TMA; m/z 76.0 → 58.1 for TMAO; and m/z 85.0 → 66.2 for d9-TMAO.
3
Quantitative Lipidomics Protocol for SARS-CoV-2
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Concentrated samples (500 µL) were denatured with 500 µL of LC‐MS–grade MeOH containing the internal standards then warmed at 60º Celsius for 30 minutes to inactivate SARS‐CoV‐2, denatured overnight (−20° Celsius), and then centrifuged (10,000 g) to remove the denaturated proteins. Under these conditions, no significant degradation of internal standards and selected lipids and lipid mediators was observed (Figure S1). Supernatants were diluted with water containing 0.01% acetic acid and lipids were extracted by solid phase extraction using Strata‐X cartridges (Polymeric Reversed Phase, Phenomenex, USA) as described before.12 Lipids were separated using the same column and LC program as in,12 quantification was done using a Shimadzu 8050 triple quadrupole mass spectrometer and analyses were performed using multiple reaction monitoring for the specific mass transitions of each lipid as well as the deuterated internal standard used (Table S2). Due to the large number of analytes being quantitated and/or commercial availability, some lipid mediators were analyzed with a surrogate, deuterated internal standard.
4
Quantitative Analysis of Gut Metabolites
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Stable isotope dilution high-performance liquid chromatography with on-line tandem mass spectrometry (LC-MS/MS) was used for quantification of levels of TMAO, TMA, choline, carnitine, betaine, and γ-butyrobetaine in plasma, as previously described (Wang et al., 2014a (link)). Their d9(methyl) isotopologues were used as internal standards. LC-MS/MS analyses were performed on a Shimadzu 8050 triple quadrupole mass spectrometer. IMC and d2-IMC, along with other metabolites, were monitored using multiple reaction monitoring of precursor and characteristic product ions as follows: m/z 230.0 → 58.0 for IMC; m/z 232.0 → 60.1 for d2-IMC; m/z 76.0 → 58.1 for TMAO; m/z 85.0 → 66.2 for d9-TMAO; m/z 60.2 → 44.2 for TMA; m/z 69.0 → 49.1 for d9-TMA; m/z 104.0 → 60.1 for choline; m/z 113.1 → 69.2 for d9-choline; m/z 118.0 → 58.1 for betaine; m/z 127.0 → 66.2 for d9-betaine.
5
Quantification of Metabolites by LC-MS/MS
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Levels of endogenous and isotope labeled γBB and TMAO were determined as previously described by stable isotope dilution liquid chromatography–tandem mass spectrometry (LC-MS/MS) in positive ion multiple reaction monitoring (MRM) mode using a Shimadzu 8050 triple quadrupole mass spectrometer with ultra-HPLC interface.6 (link),8 (link) High-sensitivity C-reactive protein, plasma glucose, hemoglobin A1C, creatinine and lipid profiles were measured on the Roche Cobas platform (Roche Diagnostics). Laboratory personnel performing clinical and mass spectrometry analyses were blinded to sample group allocation and clinical data during analysis.
6
TMAO Quantification Protocol Using LC-MS/MS
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Fasting blood samples were collected in EDTA tubes, processed to isolate plasma, and frozen at −80 °C until analysis. TMAO levels were measured using stable isotope dilution liquid chromatography with online tandem mass spectrometry using an 8050 triple quadrupole mass spectrometer (Shimadzu Scientific Instruments, Columbia, MD, USA) as previously described [1 (link),31 (link)]. The TMAO assay showed good inter- and intraday stability (CVs < 7%), accuracy (>98.5%), and stability across freeze–thaw cycles (intercycle CV < 9%) [31 (link)].
7
Lipid Extraction and Quantification of eCBome Mediators
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Lipids were extracted from tissue samples according to the Bligh and Dyer method [21 (link)], as previously described [16 (link),20 (link)].
The quantification of eCBome-related mediators (Supplementary Table S1 ), was carried out by HPLC interfaced with the electrospray source of a Shimadzu 8050 triple quadrupole mass spectrometer using multiple reaction monitoring in positive ion mode for the compounds and their deuterated homologs. In the case of unsaturated monoacylglycerols, the data are presented as 2-monoacylglycerols (2-MAGs) but represent the combined signals from the 2- and 1(3)-isomers since the latter are most likely generated from the former via acyl migration from the sn-2 to the sn-1 or sn-3 position.
The quantification of eCBome-related mediators (
8
Analysis of Trace-Level Drugs in Wastewater
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A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was employed to analyze low (ppt to ppb) drug levels in extracted wastewater influent samples. A complete list of all targeted compounds is located in SI Tables 1 and 2. All LC-MS/MS data were collected using a Shimadzu 8050 triple quadrupole mass spectrometer, operating in the positive electrospray mode (ESI+), coupled to a Nexera HPLC (Shimadzu; Kyoto, Japan). Analyte separations were achieved using a Phenomenex (Torrance, CA) Kinetex Phenyl-Hexyl reversed phase HPLC column (50 × 4.6 mm, 2.6 μm), and corresponding guard column. The HPLC column, and guard column, were kept at 40 °C throughout all chromatographic runs. Mobile phase A was 0.1% ammonium formate in water and mobile phase B was 0.1% FA in MeOH. An injection volume of 15 μL and total flow rate of 0.700 mL min−1 were used. Retention times, precursor ions, and product ions monitored for the non-labeled compounds are listed in SI Table 1, and the labeled compounds are listed in SI Table 2. More details, on the LC-MS/MS method can be found in the SI file.
9
Quantifying Endocannabinoid Metabolome
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eCBome mediators were extracted from V1 and V7 plasma samples of a total of 23 participants (13 placebo and 10 CAE). Sample processing and LC–MS/MS parameters were exactly as described by Turcotte and collaborators [47 (link)]. The quantification of eCBome-related mediators (Table S2 ) was carried out by an HPLC system interfaced with the electrospray source of a Shimadzu 8050 triple quadrupole mass spectrometer and using multiple reaction monitoring in positive ion mode for the compounds and their deuterated homologs.
In the case of unsaturated monoacylglycerols, the data are presented as 1/2-monoacylglycerols (2-MAGs) but represent the combined signals from the 2- and 1(3)-isomers since the latter were most likely generated from the former via acyl migration from the sn-2 to the sn-1 or sn-3 position.
In the case of unsaturated monoacylglycerols, the data are presented as 1/2-monoacylglycerols (2-MAGs) but represent the combined signals from the 2- and 1(3)-isomers since the latter were most likely generated from the former via acyl migration from the sn-2 to the sn-1 or sn-3 position.
10
Gnotobiotic communities and stable isotope
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