Plasma and tissue samples from all drug-treated animals at selected time points were analysed using a previously developed non-validated LC-MS/MS method. A sensitive and highly selective liquid chromatography-tandem mass spectrometry (LC-MS) method was used to determine the drug concentration in mouse plasma samples or tissue homogenates. LC-MS analysis was carried out on a Bruker amaZon SL mass spectrometer using positive/negative ion ESI mode. Chromatographic separation was achieved on an Ascentis Express C18 column (5 cm × 2.1 mm, 2.7 µM, Supelco Technologies) at room temperature with a thermostatted column oven. A gradient elution of eluents A (acetonitrile (LiChrosolv, Reag. Ph Eur) + 0.1% formic acid (Sigma Aldrich, 98–100%)) and B (water +0.1% formic acid) was used for separation. The flow rate was set at 1 ml/min. The injection volume was 20 µL, and the time of injection was 4 min.
Amazon sl mass spectrometer
Manufactured by Bruker
Sourced in Germany
The AmaZon SL mass spectrometer is a high-performance liquid chromatography-mass spectrometry (LC-MS) system designed for analytical and research applications. It features a compact and modular design, providing users with a versatile and reliable solution for a wide range of analytical tasks.
Automatically generated - may contain errors
Lab products found in correlation
Dataanalysis 4, by Bruker (3 mentions)
Lc 20a system, by Shimadzu (2 mentions)
Spectrometer, by Bruker (2 mentions)
Direct yellow 59, by Merck Group (2 mentions)
Formic acid, by Merck Group (1 mentions)
Silica gel 60n, by Kanto Chemical (1 mentions)
Eca 500 spectrometer, by JEOL (1 mentions)
Solarix spectrometer, by Bruker (1 mentions)
V 650 spectrometer, by Jasco (1 mentions)
Tlc silica gel 60 f254, by Merck Group (1 mentions)
Ls45 luminescence spectrophotometer, by PerkinElmer (1 mentions)
X pert pro, by Malvern Panalytical (1 mentions)
Hplc system, by Shimadzu (1 mentions)
Luna c18 2, by Phenomenex (1 mentions)
717 plus autosampler, by Waters Corporation (1 mentions)
Merck silica gel 60f 254 glass plates, by Merck Group (1 mentions)
Linomat, by CAMAG (1 mentions)
Dataanalysis, by Bruker (1 mentions)
Trap control, by Bruker (1 mentions)
2998 photodiode array detector, by Waters Corporation (1 mentions)
16 protocols using amazon sl mass spectrometer
1
Quantification of Drug Concentrations in Plasma and Tissues
2
Characterization of Organic Compounds
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Chemical reagents were purchased from Tokyo Chemical Industry Co., Ltd., FUJIFILM Wako Pure Chemical Corporation, Watanabe Chemical Industries, Ltd. and Bachem Holding AG., and used without further purification. Thin-layer chromatography (TLC) was performed on TLC silica gel 60F254 (Merck). Column chromatography was performed on silica gel 60N (Kanto Chemical Co., Inc., spherical neutral, 63 to 210 µm). 1H and 13C NMR spectra were recorded on a JEOL ECA500 spectrometer in CDCl3, CD3OD or dimethyl sulfoxide-d6 (DMSO-d6) with tetramethylsilane (TMS) or residual non-deuterated solvents as the internal references. Multiplicities are abbreviated as follows: s = singlet, d = doublet, t = triplet, q = quartet, m = multiplet, dd = double doublet, and br = broad. LRMS (ESI-MS) analysis was conducted using a Bruker amaZon SL mass spectrometer. HRMS (ESI-FT-ICR-MS) analyses were conducted using a Bruker Solarix spectrometer. The absorption spectra were measured using a Jasco V-650 spectrometer equipped with an ETCS-761 temperature controller. FT-IR spectra were recorded on a JEOL FT/IR-4100 spectrometer using KBr pellets in the range of 4000 to 400 cm−1.
3
Synthesis and Characterization of Methacrylate Citrate Polymers
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Methacrylate poly(1,12-dodecamethylene citrate) (mPDC) and Methacrylate poly(1,8-octanediol
citrate) (mPOC) was synthesized by following our previously reported protocol [17 ]. Briefly, citric acid and 1,12-dodecanediol at molar ratio of 2:3 or citric acid and 1,8-octanediol at molar ratio of 1:1 were melted (165 °C, 22 min), co-polymerized (140 °C, 60 min), purified, and freeze-dried to yield PDC of POC pre-polymer. Every 22 g PDC or POC pre-polymer was dissolved in tetrahydrofuran (180 mL) with imidazole (816 mg) and glycidyl methacrylate (17.8 mL), reacted (60 °C, 6 h), purified, and freeze-dried to yield mPDC or mPOC pre-polymer.
The characterization of PDC and POC pre-polymer was performed by a Bruker AmaZon SL Mass Spectrometer equipped with an Electrospray Ionization source (ESI) and an Ion Trap Mass Analyzer. The sample analysis was done with direct loop injection, using ethanol as the carrier solvent/mobile phase. The acquisition method was set up to perform an Alternate Positive and Negative ion acquisition, collecting both positive and negative ion information. The characterization of mPDC or mPOC pre-polymer was performed by proton nuclear magnetic resonance (1H NMR) using deuterated dimethyl sulfoxide (DMSO‑d6) as the carrier solvent.
citrate) (mPOC) was synthesized by following our previously reported protocol [17 ]. Briefly, citric acid and 1,12-dodecanediol at molar ratio of 2:3 or citric acid and 1,8-octanediol at molar ratio of 1:1 were melted (165 °C, 22 min), co-polymerized (140 °C, 60 min), purified, and freeze-dried to yield PDC of POC pre-polymer. Every 22 g PDC or POC pre-polymer was dissolved in tetrahydrofuran (180 mL) with imidazole (816 mg) and glycidyl methacrylate (17.8 mL), reacted (60 °C, 6 h), purified, and freeze-dried to yield mPDC or mPOC pre-polymer.
The characterization of PDC and POC pre-polymer was performed by a Bruker AmaZon SL Mass Spectrometer equipped with an Electrospray Ionization source (ESI) and an Ion Trap Mass Analyzer. The sample analysis was done with direct loop injection, using ethanol as the carrier solvent/mobile phase. The acquisition method was set up to perform an Alternate Positive and Negative ion acquisition, collecting both positive and negative ion information. The characterization of mPDC or mPOC pre-polymer was performed by proton nuclear magnetic resonance (1H NMR) using deuterated dimethyl sulfoxide (DMSO‑d6) as the carrier solvent.
4
Spectroscopic and Mass Spectrometric Characterization of Compounds
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All chemicals were purchased from commercial sources unless otherwise specified. All the solvents were of analytical reagent grade and were used without further purification. All oligonucleotides used in this work were synthesized and purified by Shanghai Sangon Biotechnology Co. Ltd. (Shanghai, China). Their sequences were given in Table S1. Fluorescence studies were performed on a LS45 luminescence spectrophotometer (Perkin-Elmer, USA). A quartz cuvette with 2 mm x 2 mm path length was used for the spectra recorded at 10 nm slit width for both excitation and emission unless otherwise specified. Mass spectra (MS) were recorded on Bruker amaZon SL mass spectrometer with an ESI or ACPI mass selective detector.
1 H and 13 C NMR spectra were recorded using TMS as the internal standard in CDCl3 or DMSO-d6 with a Bruker BioSpin GmbH spectrometer at 400 MHz and 100 MHz, respectively.
The purities of synthesized compounds were confirmed by using analytical HPLC with a dual pump Shimadzu LC-20A system equipped with a photo-diode array detector and a C18 column (250 mm x 4.6 mm, 5 µM YMC) and eluted with acetonitrile/water (47:53) containing 0.5 % acetic acid at a flow rate of 1.0 mL/min. The stock solutions of the ligands were prepared at 5 mM with DMSO. The stock solution was then diluted to the required concentration with Tris-HCl buffer containing 60 mM KCl for experiments.
1 H and 13 C NMR spectra were recorded using TMS as the internal standard in CDCl3 or DMSO-d6 with a Bruker BioSpin GmbH spectrometer at 400 MHz and 100 MHz, respectively.
The purities of synthesized compounds were confirmed by using analytical HPLC with a dual pump Shimadzu LC-20A system equipped with a photo-diode array detector and a C18 column (250 mm x 4.6 mm, 5 µM YMC) and eluted with acetonitrile/water (47:53) containing 0.5 % acetic acid at a flow rate of 1.0 mL/min. The stock solutions of the ligands were prepared at 5 mM with DMSO. The stock solution was then diluted to the required concentration with Tris-HCl buffer containing 60 mM KCl for experiments.
5
Lipid Fractionation and ESI-IT MS Analysis
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Dried lipid extracts were dissolved in chloroform and automatically spotted onto a normal phase HPTLC glass plate (Merck KGaA, Darmstadt, Germany) by a Linomat device (CAMAG, Muttenz, Switzerland). Chloroform/ethanol/water/triethylamine (30:35:7:35, by vol.) was used as the mobile phase. After drying at room conditions for 15 min, the separated lipid fractions were visualized by dipping the entire plate into primuline (Direct Yellow 59, Sigma-Aldrich, Taufkirchen, Germany) solution (50 mg/l in acetone/water (80:20, by vol.)). The lipids in each spot were automatically eluted by a Plate Express™ TLC plate reader (Advion, Ithaca, NY, USA) with methanol as solvent and analyzed by direct transfer into the ESI-IT mass spectrometer.
ESI-IT MS was performed on an Amazon SL mass spectrometer (Bruker Daltonik GmbH) by direct infusion. Conditions were the following: spray voltage 4.5 kV, end plate offset 500 V, nebulizer gas 7 psi, drying gas (N2) 3 l/min, capillary temperature 180°C, flow rate 3 μL/min, sheath gas (He) flow rate 25 a.U. Spectra were recorded in the enhanced resolution mode by positive or negative ionization with a maximum ionization time of 50 ms.
ESI-IT MS was performed on an Amazon SL mass spectrometer (Bruker Daltonik GmbH) by direct infusion. Conditions were the following: spray voltage 4.5 kV, end plate offset 500 V, nebulizer gas 7 psi, drying gas (N2) 3 l/min, capillary temperature 180°C, flow rate 3 μL/min, sheath gas (He) flow rate 25 a.U. Spectra were recorded in the enhanced resolution mode by positive or negative ionization with a maximum ionization time of 50 ms.
6
Synthesis and Characterization of CXCL8 Isoforms
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CXCL8(-2-77), CXCL8(1-77), CXCL8(2-77), CXCL8(3-77), CXCL8(6-77), CXCL8(7-77), CXCL8(8-77), and CXCL8(9-77) were chemically synthesized on an Activotec P11 automated peptide synthesizer (Activotec, Cambridge, U.K.) based on N-(9-fluorenyl) methoxycarbonyl (Fmoc) chemistry as described (28 (link)). RP – high performance liquid chromatography (HPLC) was used to purify synthesized proteins to homogeneity (Proto 300 C4 column; 150 ×4.6 mm, Higgins Analytical Inc., Mountain View, CA). Elution was performed with an acetonitrile gradient in 0.1% (v/v) trifluoroacetic acid (TFA), with two percent of the effluent being used for analysis with online electrospray ionization – ion trap mass spectrometry (AmaZon SL mass spectrometer; Bruker Daltonics, Bremen, Germany). Homogeneous CXCL8 proteins were folded into their correct configurations as reported previously (28 (link)). The purity and concentration of synthesized and correctly folded CXCL8 forms was confirmed by ion trap mass spectrometry and specific sandwich ELISAs, respectively.
7
Comprehensive Characterization of Compounds
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Solution-state 1H and 13C NMR spectra were recorded on a Bruker AVANCEIII 300 MHz spectrometer at 298 K. Deuterated solvents were obtained from Cambridge Stable Isotopes and were used as received with the chemical shifts (δ) referenced internally to the residual solvent resonances and quoted in ppm. PXRD data were collected with a PANanalytical X'Pert PRO diffractometer producing Cu-Kα (λ = 1.5406 Å) radiation and equipped with a solid state PIXcel detector. Mass spectrometry was performed at the Mass Spectrometry Analysis Facility at the University of Sydney on a Bruker amaZon SL mass spectrometer. Elemental microanalysis was carried out at the Chemical Analysis Facility – Element Analysis Service in the Department of Chemistry and Biomolecular Science at Macquarie University, Australia.
8
Metabolomic Analysis of Crude Extracts by HPLC-MS/MS
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For metabolomic content evaluation, all crude extracts were evaluated by HPLC-MS/MS. The analyses were developed on an HPLC system (Shimadzu) coupled to an AmaZon SL mass spectrometer (Bruker Daltonics) fitted with an electrospray ionization source operating in positive ionization mode and with an ion trap MS detector. The chromatographic separation occurred on a Phenomenex Luna C18(2) (5 μm, 250 mm × 4.6 mm) column, using a gradient from 5 to 100% MeOH over 35 min followed by 100% MeOH for 7 min, with a flow rate of 1.0 mL/min. Both solvents contained 0.1% formic acid. The column temperature was set to 40°C. (Ion source: ESI, voltage: 3,500 V, capillary temp.: 310 °C, m/z range: 100–1,200, gas pressure: 40 psi.) An untargeted method was employed for acquisition of MS/MS spectra, in which the analyzer selects in each MS scan the three highest-intensity ions to fragment, using a ramp of collision energy from 20 to 75 eV.
9
HPLC-ESI-MS Analysis of MAAs
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The filtered samples of MAAs (dissolved in water) were analyzed with a HPLC system (Waters 2998, Photodiode Array, pump L-7100, USA) equipped with a Licrospher RP 18 column and guard 5 μm packing; 250 mm×4 mm inside diameter). The samples (10 μL) were injected into the HPLC column through a Waters 717 plus autosampler. Acetic acid (0.02%) in doubledistilled water was used as a mobile phase, which was isocratically run at a flow rate of 1 mL.min -1 . Absorption spectra of separated peaks were recorded for each second between 250 and 400 nm directly on the HPLC system. Purified MAAs collected by HPLC were used to produce protonated molecules by ESI-MS were recorded on an Amazon SL mass spectrometer (Bruker Daltonics Inc., Bremen, Germany). Cone voltage of 30V was found to induce the formation of (M+H) 1+ with a mass range of 100-1,000 m/z. Other MS settings were the following: capillary voltage (5,500 V) and temperature (300°C). The m/z scale of the MS was calibrated using the external calibration standard electrospray 'tuning mix' from Agilent Technologies (Santa Rosa, USA). Data were analyzed using the software Data Analysis 4.0 (Bruker Daltonics Inc., Bremen, Germany).
10
Analytical Characterization of Organic Compounds
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Melting points (m.p.) were determined using a SRS Opti Mel automated melting point instrument without correction. 1H and 13C NMR spectra were recorded using TMS as the internal standard in DMSO-d6 with a Bruker BioSpin GmbH spectrometer at 400 and 100 MHz, respectively. Mass spectra (MS) were recorded on Bruker amaZon SL mass spectrometer with an ESI or ACPI mass selective detector. Reactions progress and compounds were checked by TLC with Merck silica gel 60F-254 glass plates. All chemicals were purchased from commercial sources unless otherwise specified, and all the solvents were analytical grade. The purities of synthesized compounds were confirmed by HPLC with a dual pump Shimadzu LC-20A system equipped with a photo-diode array detector and a C18 column (250 × 4.6 mm, 5 μM YMC) and eluted with acetonitrile/water (47:53) containing 0.5% acetic acid at flow rate of 1.0 ml/min.
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