Blood samples were collected via radial or femoral access before heparinization using vacutainer tubes containing EDTA. Samples were maintained at 4°C, processed within 3 h, and then stored at −80°C until further analysis. Plasma levels of TMAO were quantified by stable isotope dilution high-performance liquid chromatography with online electrospray ionization tandem mass spectrometry using an API 3200 triple quadrupole mass spectrometer (AB SCIEX, Framingham, MA) with a d9-(trimethyl)-labeled internal standard as described previously (13 (link)). The estimated glomerular filtration rate (mL/min per 1.73 m2) was calculated using the Modification of Diet in Renal Disease study equation (14 (link)).
Api 3200 triple quadrupole mass spectrometer
Manufactured by AB Sciex
Sourced in Germany, Canada, United States
The API 3200 is a triple quadrupole mass spectrometer designed for qualitative and quantitative analysis. It features high sensitivity, fast scanning, and precise mass selection capabilities. The instrument is capable of performing multiple reaction monitoring (MRM) experiments to identify and measure target analytes in complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Vortex genie 2, by Scientific Industries (2 mentions)
Zorbax rrhd eclipse plus c18 column, by Agilent Technologies (2 mentions)
Agilent 1290 infinity lc, by Agilent Technologies (2 mentions)
Universal 320r centrifuge, by Hettich (2 mentions)
Synapt g2 s mass spectrometer, by Waters Corporation (1 mentions)
Cdd 6a, by Shimadzu (1 mentions)
Lc 10advp hplc, by Shimadzu (1 mentions)
Agilent 1290 lc system, by Agilent Technologies (1 mentions)
Analyst 1, by AB Sciex (1 mentions)
Dmye00b, by R&D Systems (1 mentions)
Analyst software, by AB Sciex (1 mentions)
Nicolet is10 ft ir spectrometer, by Thermo Fisher Scientific (1 mentions)
Drx 600 nmr spectrometer, by Bruker (1 mentions)
Prism 6, by GraphPad (1 mentions)
Kinetex c18 column, by Phenomenex (1 mentions)
Agilent 1200 series, by Agilent Technologies (1 mentions)
1200 series hplc system, by Agilent Technologies (1 mentions)
20 protocols using api 3200 triple quadrupole mass spectrometer
1
Quantifying Plasma TMAO Levels
2
Plasma TMAO, Cardiac Biomarkers in Hospitals
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Peripheral blood samples were collected before heparinization using vacutainer tubes containing ethylenediamine tetra-acetic acid, maintained at 4°C and processed within 3 h. Then, plasma samples were stored at −80°C and thawed once for analysis. Plasma levels of TMAO were quantified by stable isotope dilution high performance liquid chromatography with online tandem electrospray ionization mass spectrometry using an API 3200 triple quadrupole mass spectrometer (AB SCIEX, Framingham, MA) with a d9-(trimethyl)-labeled internal standard as described previously (37 (link)). In addition, cardiac troponin I (cTnI) and NT-proBNP were measured as routine biochemical work-up during the hospital stay. As a standard care, cTnI was measured at least 3 times within 24 h after admission and daily until normalization and then every 2 days before discharge; NT-proBNP was measured on admission, immediately after and on the second day after the index procedure and then up to physicians. Results of cTnI and NT-proBNP during hospitalization were retrieved from electronic medical records and baseline values and peak values were reported.
3
TMAO Quantification in Plasma
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Blood samples were collected from radial or femoral access before heparinization during angiography using vacutainer tubes containing EDTA. Then the blood sample was centrifuged and stored at -80 °C. API 3200 triple quadrupole mass spectrometer (AB SCIEX, USA) was used to measure plasma TMAO according to the manufacturer’s instructions [19 (link)].
4
Measurement of Plasma TMAO and Cardiac Biomarkers
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Blood samples were gathered into tubes with ethylenediaminetetraacetic acid through radial or femoral artery before percutaneous coronary intervention (PCI). These were processed at 4°C within 3 h, and then stored at −80°C until further analysis. As mentioned previously, the stable isotope dilution high‐performance liquid chromatography with online electrospray ionization tandem mass spectrometry was used to measure plasma TMAO levels, using an API 3200 triple quadrupole mass spectrometer (AB SCIEX, Framingham, Massachusetts) with a d9(trimethyl)‐labelled internal standard.20 HsCRP testing was routinely collected via cubital or basilic veins in an overnight fasting state on the day after the PCI procedure and measured using an immunoturbidimetric assay (Beckmann Assay, Bera, California). The other blood test indicators are routinely detected in the hospital central laboratory. The N‐terminal pro‐B‐type natriuretic peptide (NT‐proBNP) and cardiac troponin I (cTnI) levels were measured several times during hospitalization, and the baseline and peak values were recorded.
5
UHPLC-MS/MS Analysis of Compounds
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UHPLC-MS/MS analyses were performed in an Agilent 1290 Infinity LC (Agilent Technologies, Waldbronn, Germany) coupled to an API 3200 triple quadrupole mass spectrometer (AB Sciex, Darmstadt, Germany) with electrospray ionization (ESI). The chromatographic separation was performed using an Agilent Zorbax Eclipse Plus RRHD C18 column (50 × 2.1 mm, 1.8 µm). Analyst software (Version 1.6.3, AB Sciex, Darmstadt, Germany) was used for acquisition and data analysis.
During the sample treatment, an evaporator System (System EVA-EC, from VLM GmbH, Bielefeld, Germany), a vortex-2 Genie (Scientific Industries, Bohemia, NY, USA), a universal 320R centrifuge (Hettich ZENtrifugen, Tuttlingen, Germany), and a kitchen blender were used.
During the sample treatment, an evaporator System (System EVA-EC, from VLM GmbH, Bielefeld, Germany), a vortex-2 Genie (Scientific Industries, Bohemia, NY, USA), a universal 320R centrifuge (Hettich ZENtrifugen, Tuttlingen, Germany), and a kitchen blender were used.
6
Plasma Biomarkers of Cardiometabolic Risk
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Blood samples were taken from the radial or femoral artery before heparinization and percutaneous coronary intervention (PCI) at admission (V1) and the cubital vein approximately one month later when patients came to the clinic (V2). The samples were maintained with tubes containing ethylenediaminetetraacetic at 4 °C, centrifuged within 3 h, the supernatant was separated and stored at −80 °C until subsequent analysis. As described in previous studies, plasma levels of TMAO and its precursors were measured by stable isotope dilution high-performance liquid chromatography with online electrospray ionization tandem mass spectrometry using an API 3200 triple quadrupole mass spectrometer (AB SCIEX, Framingham, MA, USA) with a d9-TMAO, d9-betaine, d9-choline, and d3-carnitine internal standard [22 (link),23 (link)]. The other blood parameters were routinely measured in the hospital’s central laboratory.
7
Phospholipid Profiling via HPLC-MS
8
Quantifying PFOS and OS in Samples
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A nitrogen evaporator was used to remove the organic solvents in the samples, and the residues were dissolved in methanol to reach constant volume. All PFOS and OS concentrations were measured using external standard method, and their calibration curves are shown in Figure S17 . High PFOS concentrations (>1 mg/L) and OS concentration were determined by a LC-10ADvp HPLC with a CDD-6A conductivity detector from Shimadzu (Japan). A mixture of methanol/0.02 mol/L NaH2PO4 solution (70/30, v/v) was selected as the mobile phase, and the limit of detection (LOD) was 0.149 mg/L for PFOS and 0.033 mg/L for OS. Low PFOS concentrations (<1 mg/L) were determined by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) with an UltiMate 3000 HPLC (Dionex by Thermo Fisher Scientific Inc., MA, USA) equipped with an API 3200 triple quadrupole mass spectrometer (AB SCIEX, ON, Canada). The mobile phase consisted of a binary mixture of 10 mmol/L ammonium acetate and methanol with a total flow rate of 0.3 mL/min. The LOD was measured to be 0.112 μg/L.
9
Quantification of Artemisinin in A. annua
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Artemisinin was quantified using UHPLC-MS/MS analysis performed on an Agilent 1290 LC system (Agilent, Waldbronn, Germany) connected to an API 3200 triple quadrupole mass spectrometer by a TurboIon source (AB Sciex, Darmstadt, Germany). Artemisinin was separated on a Nucleoshell C18 column (2.6 µm, 50 x 3 mm; Macherey-Nagel, Düren, Germany) at a flow rate of 500 µl min-1 using 0.02% (v/v) acetic acid in water or in acetonitrile as eluents A and B, respectively. The ion source was operated in the positive mode, and data were acquired in the multiple reaction monitoring mode with target scan time of 50 ms. The IntelliQuant algorithm of the Analyst 1.6.2 software (AB Sciex, Darmstadt, Germany) was used to integrate the peaks for artemisinin. Concentrations were calculated using an artemisinin standard curve in the range of 0 to 5.6 µg ml-1 and divided by the dry weights. Our full protocol for determining artemisinin level in A. annua was recently published (Paponov et al., 2023 (link)).
10
Quantification of Plasma TMAO and MPO
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Blood samples were collected via radial or femoral access before the interventional procedure using vacutainer tubes containing ethylenediaminetetraacetic acid (EDTA). Samples were maintained at 4°C, processed within 3 h, and then stored at −80°C until further analysis. Plasma levels of TMAO were quantified by stable isotope dilution high-performance liquid chromatography with online electrospray ionization tandem mass spectrometry using an API 3200 triple quadrupole mass spectrometer (AB SCIEX, Framingham, MA) with a d9-(trimethyl)-labeled internal standard as described previously. (Wang et al., 2014a (link)). Plasma MPO was measured by enzyme-linked immunosorbent assay using a commercial kit (DMYE00B, R&D Systems, United States), according to procedures recommended by the manufacturer. Duplicates of 20 randomly selected samples and all samples were measured to evaluate the intra-assay coefficient variation of TMAO and MPO, respectively. The estimated glomerular filtration rate (eGFR; ml/min per 1.73 m2) was calculated using the Modification of Diet in Renal Disease study equation (Levey et al., 2009 (link)).
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