20-HETE was quantified by LC-MS/MS-based lipidomics. Blood was withdrawn at the end of the experiment by aspiration from the inferior vena cava into a heparinized syringe. Urine and plasma samples were mixed with 2 volumes of cold methanol, internal standards were added and samples kept at −80°C. Renal preglomerular arteries and mesenteric arteries were microdissected and incubated in Kreb’s bicarbonate buffer, pH 7, 1 mM NADPH at 37°C for 1 h. Tissue incubations were terminated with 2 volumes of cold methanol, internal standards were added, and samples were kept at −80°C. 20-HETE was extracted and levels quantified by LC-MS/MS on a Shimadzu Triple Quadrupole Mass Spectrometer LCMS-8050 as previously described. [41 (link), 42 (link)]
Lcms 8050 triple quadrupole mass spectrometer
Manufactured by Shimadzu
Sourced in Japan
The LCMS-8050 is a triple quadrupole mass spectrometer manufactured by Shimadzu. It is designed for high-sensitivity and high-speed quantitative analysis of small molecules. The LCMS-8050 features a UF-Qarray technology that enhances ion transmission and improves signal-to-noise ratio. It also includes a heated ESI probe and dual ion source to provide a wide range of ionization options.
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Lab products found in correlation
Nexera x2 uhplc system, by Shimadzu (8 mentions)
Nexera uhplc system, by Shimadzu (5 mentions)
Lc ms ms system, by Shimadzu (4 mentions)
Labsolutions v5, by Shimadzu (4 mentions)
Uhplc system, by Shimadzu (3 mentions)
Lc solutions software, by Shimadzu (2 mentions)
Nexera x2 ultra hplc, by Shimadzu (2 mentions)
Sil 30ac, by Shimadzu (2 mentions)
Lc 30ad dual plunger parallel flow pumps, by Shimadzu (2 mentions)
Cbm 20a controller, by Shimadzu (2 mentions)
Nexera uhplc, by Shimadzu (2 mentions)
Labsolution, by Shimadzu (2 mentions)
Synergi 4 m max rp column, by Phenomenex (2 mentions)
Nexera uplc system, by Shimadzu (1 mentions)
Luna c18, by Phenomenex (1 mentions)
Nexera hplc system, by Shimadzu (1 mentions)
Labsolutions software, by Shimadzu (1 mentions)
Acquity uplc beh, by Waters Corporation (1 mentions)
C18 column, by Waters Corporation (1 mentions)
β glucuronidase arylsulfatase from helix pomatia, by Merck Group (1 mentions)
41 protocols using lcms 8050 triple quadrupole mass spectrometer
1
Quantification of 20-HETE by LC-MS/MS Lipidomics
2
UPLC-MS/MS Analysis of Compounds
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The UPLC-MS/MS system consisting of Shimadzu Ultra-High Performance Liquid Chromatograph LC-30A and Triple Quadrupole Mass Spectrometer (LCMS-8050, SHIMADZU, Japan).
3
LC-MS/MS Analysis of Metabolites
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LC-MS/MS analysis was performed using a Shimadzu Nexera UPLC system (Shimadzu Corp., Kyoto, Japan) with an LCMS-8050 triple quadrupole mass spectrometer (Shimadzu Corp., Kyoto, Japan). Chromatographic separation was achieved using an Intrada amino acid column (150 mm × 2 mm, 3 µm). Nebulizing and drying gas flow was 3.0 L/min and 10.0 L/min, respectively. The pressure of the collision-induced dissociation (CID) gas was 270 kPa. The interface, desolvation line (DL) and heat block temperature were set at 200 °C, 200 °C, and 300 °C, respectively. Mobile phase A was composed of ACN/tetrahydrofuran/25 mM ammonium formate/formic acid (9/75/16/0.3, v/v/v/v) and mobile phase B was composed of ACN/100 mM ammonium formate (20/80, v/v). The mobile phase gradient program, initially set at 0% of mobile phase B, increased to 17% of mobile phase B (2.5–6.5 min), increased to 100% of mobile phase B (6.5–10 min), held for 5 min, and decreased to the initial 0% of mobile phase B, followed by a 3 min re-equilibration period. The total flow rate was set at 0.6 mL/min; the column temperature was maintained at 35 °C. In SRM modes with electrospray ionization (ESI) mode, one quantitative ion for each metabolite was used for peak quantification.
4
Lipid Mediators Quantification by LC-MS/MS
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All components of LC–MS/MS system are from Shimadzu Scientific Instruments, Inc. (Columbia, MD, USA). LC system was equipped with four pumps (Pump A/B: LC-30AD, Pump C/D: LC-20AD XR), a SIL-30AC autosampler (AS), and a CTO-30A column oven containing a 2-channel six-port switching valve. The LC separation was conducted on a C8 column (Ultra C8, 150 × 2.1 mm, 3 µm, RESTEK, Manchaca, TX, USA) along with a Halo guard column (Optimize Technologies, Oregon City, OR, USA). The MS/MS analysis was performed on Shimadzu LCMS-8050 triple quadrupole mass spectrometer. The instrument was operated and optimized under both positive and negative electrospray and multiple reaction monitoring modes (± ESI MRM). The settings of flow rate and gradient program for the LC system as well as MS/MS conditions are recommended by a software method package for 158 lipid mediators (Shimadzu Scientific Instruments, Inc., Columbia, MD, USA) and further optimized following previously published quantification method [27 (link)]. All analyses and data processing were completed on Shimadzu LabSolutions V5.91 software (Shimadzu Scientific Instruments, Inc., Columbia, MD, USA).
5
Quantification of Plasma Melatonin Levels
6
LCMS-8050 Protocol for Amino Acid Quantification
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Mass spectrometric measurements were performed on a LCMS-8050 triple-quadrupole mass spectrometer (Shimadzu, Kyoto, Japan) coupled with a Nexera HPLC system (Shimadzu, Kyoto, Japan). The instrument parameters were as follows: injection volume was 4 μL, flow rate of the flow-injection analysis solvent was 0.2 mL/min, the sample cooler temperature was 25°C, rate of N2 drying gas was 10 L/min, the rate of N2 nebulizing gas was 3 L/min, and the capillary voltage was −3.0 kV for the negative ion detection mode. The signal intensity was monitored for m/z values of deprotonated molecules [M−H]− (m/z 88 for Ala, m/z 118 for Thr and m/z 164 for Phe) in the selected ion monitoring mode. The signal intensities were estimated from the peak areas of selected-ion chromatograms on flow injection analysis (triplicated measurements).
7
Comprehensive Metabolite Analysis by LC-MS/MS
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All LC-MS/MS analyses were performed using the LCMS-8050 triple quadrupole mass spectrometer coupled with the Nexera X2 UHPLC system (Shimadzu, Kyoto, Japan) and Lab Solutions software (Shimadzu, Kyoto, Japan). To analyze metabolites, analytes were separated into four groups and analyzed with each optimized method for improvement of the measurement sensitivity with each metabolite. The LC method used for each compound and optimized MS/MS conditions are summarized in Supplementary Table S1 . For all four methods, column oven temperature was set at 40 °C and electrospray ionization mode was chosen as the ion source probe. Ion source probe conditions were as follows: probe voltage, 4000 V; desolvation line temperature, 100 °C; block heater temperature, 150 °C; interface temperature, 400 °C; nebulizing gas flow, 2 L/min; drying gas flow, 3 L/min; and heating gas flow, 17 L/min. Column and mobile phase were selected for metabolites with high sensitivity, and calibration and internal standards for each group are listed in Supplementary Data S2 and Supplementary Table S2 .
8
Sensitive BPA Quantification in Plasma
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From anesthetized animals, plasma was collected by centrifugation of whole blood (800 g for 15 min). The determination of BPA was conducted as previously reported52 (link), with some minor modifications. Briefly, d16-BPA was added as internal standard and samples were defatted with hexane and then extracted with dichloromethane. The extracts were purified in two successive Solid Phase Extraction steps, one with a Florisil solid phase and one with a C18 solid phase. Deconjugation was carried out using β-Glucuronidase/Arylsulfatase from Helix pomatia (Sigma-Aldrich).
Chromatographic separations were carried out using a LCMS-8050 triple quadrupole mass spectrometer equipped with a Nexera UHPLC System (Shimadzu). BPA was separated on an Acquity UPLC BEH (2.1 mm × 50 mm, 1.7μm) C18 column (Waters). A 1.5 min linear gradient was used from 10–95% methanol in water followed by a hold at 95% for 1.0 min at a flow rate of 0.4 ml/min. Negative ion electrospray mass spectrometry with selected reaction monitoring (SRM) and a dwell time of 50 ms per transition was used for the measurement of each analyte. The SRM transitions for BPA were m/z 227.20 to 212.20 (quantifier) and m/z 227.20 to133.15 (qualifier).
Chromatographic separations were carried out using a LCMS-8050 triple quadrupole mass spectrometer equipped with a Nexera UHPLC System (Shimadzu). BPA was separated on an Acquity UPLC BEH (2.1 mm × 50 mm, 1.7μm) C18 column (Waters). A 1.5 min linear gradient was used from 10–95% methanol in water followed by a hold at 95% for 1.0 min at a flow rate of 0.4 ml/min. Negative ion electrospray mass spectrometry with selected reaction monitoring (SRM) and a dwell time of 50 ms per transition was used for the measurement of each analyte. The SRM transitions for BPA were m/z 227.20 to 212.20 (quantifier) and m/z 227.20 to133.15 (qualifier).
9
Quantitative Sphingolipid Analysis by LC-MS
10
LC-MS/MS Quantification of Metabolites
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Samples were measured using a Nexera MP System utilizing the SIL-30ACMP Multi-Plate autosampler and LCMS-8050 triple quadrupole mass spectrometer (Shimadzu Corporation, Kyoto, Japan) equipped with an electrospray ionization-positive and -negative source. The LC column was a Phenomenex Kinetex XB-C18 150 × 2.1 mm, 1.7 µm (Phenomenex, Torrance, CA, USA), and the column oven temperature was 40 °C. Samples (10 µL) were injected and eluted with a binary solvent gradient of 0.1% formic acid in water (solvent A) and 0.1% formic acid in acetonitrile (solvent B) at a flow rate of 0.4 mL/min. Separation was conducted at 40 °C according to the following gradient program: 0–0.5 min, 30% (B) to 30% (B); 0.5–3.5 min, 30% (B) to 100% (B); 3.5–5.0 min, 100% (B); and 5.01–6 min, 100(B) to 30% (B). The desolvation line temperature was 250 °C, the heat block temperature was 100 °C, drying gas flow was 15.0 L/min, and nebulizer gas flow was 3.0 L/min. The precursor ion and product ion for each MPS type are shown in Table 4 .
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