Using the Dionex Ultimate 3000 RS UHPLC with a heated electric spray ionization (ESI) source (Thermo Fisher Scientific, Waltham, MA, USA) and Q-Exactive quadrupole orbit mass spectrometer, the metabolic spectrum of the samples was measured in ESI positive and ESI negative ion modes. ACQUITY UPLC HSS T3 COLUMN (1.8 m. 2.1100 mm) used for sample separation. The elution solvent is (A) water (mic acid, v/v, containing 0.1%) and (B) acetonitrile (mic acid, v/v), and the elution gradient is 0.01 min, 5% B; 2 min, 5% B; 4 min, 30% B; 8 min, 50% B; 10 min, 80% B; 14 min, 100% B; 15 min, 100% B; 15.1 min, 5% and 16 min, 5% B. Flow rate 0.35 mL/min, column temperature 45°C. 100–1200 m/z mass range. The solution was adjusted to 70,000 for full MS scans and 17,500 for HCD MS/MS scans. The mass spectrometer functioned as follows: The capillary temperature is 320 °C, the spray voltage is 3800 volts (+) and 3000 volts (−), the sheath gas flow rate is 40 arbitrary units (+) and 35 arbitrary units (−), and the auxiliary gas flow rate is 10 arbitrary units (+) and 8 arbitrary units (−).
Dionex ultimate 3000 rs uhplc
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Dionex Ultimate 3000 RS UHPLC is a high-performance liquid chromatography system designed for ultra-high pressure liquid chromatography (UHPLC) applications. It features a modular design and can be configured with various pump, autosampler, and detector modules to meet the needs of diverse analytical workflows.
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Lab products found in correlation
4 protocols using dionex ultimate 3000 rs uhplc
1
Targeted metabolomics using UHPLC-Q-Exactive
2
Carotenoid Profiling in Human Milk and Serum
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The carotenoids from human milk and serum samples were separated using a Dionex Ultimate 3000RS U-HPLC (Thermo Fisher Scientific, Waltham, MA, USA) using the method developed by Breithaupt et al. [41 (link)] with slight modifications [37 (link)]. High-resolution mass spectrometry measurements were completed on the basis of mass accuracy and in combination with the isotopic pattern in the SigmaFit algorithm [37 (link)]. The characteristics of experimental mass spectrum and the MS2 data were compared with the data available in the literature for carotenoid identification in human milk and serum samples [42 ,43 ,44 ,45 ,46 (link),47 (link),48 (link)]. For carotenoid quantification, stock solutions of β-carotene, β-cryptoxanthin, lutein, and lycopene were prepared at a concentration of 25 mg/L. Once the exact concentration was determined (2% maximum total error), working stock solutions for external calibration curves were prepared at 5 concentration levels ranging from 0.15 to 10.0 mg/L. The content of xanthophylls and carotenes was determined in the unsaponified extract. Zeaxanthin was quantified with the calibration curve of lutein, while xanthophyll esters were quantified as their corresponding free xanthophyll [47 (link),48 (link)].
3
Rhei Radix Et Rhizoma Metabolite Profiling
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Accurately weighed LTF 10 daily prescriptions of granules, which composed of Rhei Radix Et Rhizoma 10 g, Persicae Semen 50 g, Hirudo 30 g, Notoginseng Radix Et Rhizoma powder 15 g and dissolved in 600 mL water. The sample was centrifuged and supernatant (200 μL) was taken. 20 μL of l -2-chlorophenyl alanine (0.3 mg/mL) dissolved in methanol as internal standard, and an 800 μL mixture of methanol and water (1/4, vol/vol) was added to each sample, and the tube was vortexed for 2 min. Ultrasonic extraction of all samples in the ice water bath for 30 min, and placed at − 20 °C for 2 h. Subsequently, the samples were centrifuged at 4 °C (13,000 rpm) for 10 min. The supernatants (150 μL) from each tube were filtered via 0.22 μm microfilters and transferred to LC vials for LC–MS analysis. Both electrospray ionization (ESI)-positive and ESI-negative ion modes of metabolites were analyzed using a Dionex Ultimate 3000 RS UHPLC (Thermo Fisher Scientific, Waltham, MA, USA). All ion modes were separated by an ACQUITY UPLC HSS T3 column (1.8 μm, 2.1 × 100 mm). A detailed description of the protocol was previously published [15 (link)].
4
HPLC/APCI-hrTOF-MS Metabolite Screening
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The liquid chromatograph in the HPLC/APCI-hrTOF-MS system was Dionex Ultimate 3000RS U-HPLC (Thermo Fisher Scientific, Waltham, MA, USA). Chromatographic separation was performed as described previously [18] but using 1 M ammonium acetate in water as the ion reagent and 1.00 mL/min as flow rate. A split post-column of 0.25 mL/min was introduced directly on the mass spectrometer APCI source. Mass spectrometry was performed using a micrOTOF-QII High Resolution Time-of-Flight mass spectrometer (UHR-TOF) with q-TOF geometry (Bruker Daltonics, Bremen, Germany) equipped with an APCI interface. The scan range applied was m/z 50-1500 and mass resolving power was always over 18.000 (m/m). Instrument was operated in positive ion mode . Mass spectra and data were acquired through broad-band Collision Induced Dissociation bbCID mode, providing MS and MS/MS spectra, simultaneously . All data were used to perform multitarget-screening using TargetAnalysis TM 1.2 software (Bruker Daltonics, Bremen, Germany). Collision energy was estimated dynamically based on appropriate values for the mass and stepped across a +/-10% magnitude range to ensure good quality fragmentation spectra. The instrument control was performed using Bruker Daltonics HyStar 3.2.
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