The samples positive for oxytetracycline residues in milk using ELISA were confirmed using Ultra-HPLC-Diode Array Detector (UHPLC-DAD). The UltiMate™ 3000 Rapid Separation UHPLC system (Thermo Fisher Scientific, USA) equipped with a DAD-3000RS (deuterium and tungsten light source) and a Chromeleon™ CDS version 6 software for data analysis were used. The Acclaim™ C18 column with 4.6×250 mm size, 120 Å pore size, and 5 µm internal diameters was used for the separation of analytes.
Ultimate 3000 rapid separation uhplc system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Ultimate 3000 Rapid Separation UHPLC system is a high-performance liquid chromatography (HPLC) instrument designed for rapid and efficient separation of complex samples. It features a modular design and advanced technology to provide reliable and consistent results.
Automatically generated - may contain errors
Lab products found in correlation
Q exactive hf mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Q exactive hf orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Tsq quantiva mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Q exactive orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
C18 analytical column, by Thermo Fisher Scientific (1 mentions)
6 protocols using ultimate 3000 rapid separation uhplc system
1
UHPLC-DAD Confirmation of Oxytetracycline in Milk
2
Peptide Separation using UHPLC-Orbitrap MS
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Peptide separation was performed using an Ultimate 3000 Rapid Separation UHPLC system (Thermo Scientific, Dionex, Amsterdam, The Netherlands). The analytical column was a PepSep C18 (1.9 μm, 120 Å, ID 75 μm × 150 mm). Samples were desalted using an online C18 trapping column. Elution was performed using a linear gradient from 5% to 35% ACN with 0.1 FA in 90 min with a flow rate of 300 nL per minute. The UHPLC system coupled to a Q-Exactive HF Orbitrap mass spectrometer from Thermo Scientific was used for analysis. Data dependent acquisition was as follows: full MS scan from 250 to 1250 m/z at a resolution of 120,000. MS/MS scans of the top 15 most intense ions were followed at a resolution of 15,000.
3
Quantitative Analysis of Intracellular Creatine
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MOLM14 and MV4-11 cells were treated with different concentrations of AC220 or Ara-C for 24 or 48 hours. 1 Â 10 6 cells per condition were washed with cold PBS, and metabolites were extracted from cell pellets (intracellular) in HPLC water after at least three freeze-thaw cycles. LC/MS analysis was performed on TSQ Quantiva Mass Spectrometer and Ultimate 3000 Rapid Separation UHPLC system (both from Thermo Fisher). Chromatographic analyses were conducted using 5 mL of sample injected onto a Kinetex 1.7 mm C18 column (50 Â 2.1 mm, 100 Å particle size; Phenomenex) with security guard ULTRA cartridges (UHPLC C18 2.1 mm ID). The mobile phase consisted of 0.1% formic acid in water (solvent A) and 0.1% formic acid in acetonitrile (solvent B). The mass spectrometer was operated in SRM-positive ion mode. Spectra were monitored at m/z 132.16 > 90.1 (quantify ion) and 132.16 > 44.2 (reference ion) for creatine and m/z 135.16 > 93.1 (quantify ion) and 135.16 > 47.2 (reference ion) for creatine-d3 with dwell time at 10 ms, Q1 at 0.7, Q3 at 0.7, and CID at 1.5 mTorr. The concentrations of intracellular creatine were normalized to total protein content determined by the Qubit Protein Assay Kit.
4
Rapid Liquid Chromatography-Mass Spectrometry
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A Thermo Scientific Q-Exactive Orbitrap Mass Spectrometer (San Jose, CA, USA) was interfaced with a Thermo Scientific UltiMate 3000 Rapid Separation UHPLC system using a pneumatic assisted heated electrospray ion source. The chromatography was achieved using a gradient mobile phase along with a C8 column (Thermo Biobasic 100 × 1 mm) with a particle size of 5 μm. The initial mobile phase condition consisted of acetonitrile and water (both fortified with 0.1% of formic acid) at a ratio of 5:95. From 0 to 1 min, the ratio was maintained at 5:95. From 2 to 62 min, a linear gradient was applied up to a ratio of 50:50 and maintained for 3 min. The mobile phase composition ratio was reverted at the initial conditions and the column was allowed to re-equilibrate for 15 minutes for a total run time of 80 minutes. The flow rate was fixed at 75 µL/min and 2 µL of samples were injected. MS detection was performed in positive ion mode and operating in scan mode at highresolution, and accurate-mass (HRAM). The default scan range was set to m/z 400-1500. Data was acquired at a resolving power of 140,000 FWHM (or full width at half maximum) using automatic gain control target of 3.0x106 and maximum ion injection time of 200 msec [17] .
5
Peptide Separation and Mass Spectrometry
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Peptide separation was performed on a Thermo Scientific Ultimate 3000 Rapid Separation UHPLC system (Dionex, Amsterdam, NL) equipped with a PepSep C18 analytical column (15 cm, ID 75 μm, 1,9 μm Reprosil, 120 Å). Peptide samples were first desalted on an online installed C18 trapping column. After desalting, peptides were separated on the analytical column with a 90 min linear gradient from 5% to 35% Acetonitrile (ACN) (Biosolve) with 0.1% FA at 300 nL/min flow rate. The UHPLC system was coupled to a Q Exactive HF mass spectrometer (Thermo Fisher Scientific, San Jose, USA). Data dependent acquisition (DDA) settings were as follows: Full MS scan between 250 and 1250 m/z at resolution of 120,000 followed by MS/MS scans of the top 15 most intense ions at a resolution of 15,000.
6
High-resolution LC-MS/MS analysis of peptides
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An aliquot of 10 µL of the sample was used for the LC-MS/MS analysis as previously described [22 (link)]. In short, an online installed C18 trapping column was used for desalting. This was performed on a Thermo Scientific (Dionex, Sunnyvale, CA, USA) Ultimate 3000 Rapid Separation UHPLC system. and a PepSep C18 analytical column (15 cm, ID 75 µm, 1.9 µm Reprosil, 120 Å) was used to separate the peptides with a flowrate of 300 nL/min, solvent A (100% H2O, 0.1% FA) and solvent B (80% ACN, 0.1% FA) and a 110 min linear gradient from 5% to 32% ACN with 0.1% FA as follows: 0–3 min 0% B; 3 min 5% B; 103 min 27.5% B; 113 min 40% B; 114–120 min 95% B; 121–140 min 3% B. The UHPLC system was coupled to a Q ExactiveTM HF mass spectrometer (Thermo Scientific) with Nanospray Flex source. Mass spectra were acquired in positive ionization mode, with a full MS scan between 250–1250 m/z at resolution of 120,000 followed by MS/MS scans of the top 15 most intense ions at a resolution of 15,000 in DDA mode.
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