D-glucarate, 5-keto-4-D-deoxyglucarate and α-ketoglutarate semialdehyde were separated by HPLC, using an Ultimate-3000 HPLC system (Dionex, Idstein, Germany), equipped with an autosampler (WPS 3000TRS), a column compartment (TCC3000RS) and a diode array detector (DAD 3000RS). The column Metrosep A Supp10–250/40 column (250 mm, particle size 4.6 mm; Metrohm, Filderstadt, Germany) at 65 °C was used for separation by isocratic elution with 30 mM ammonium bicarbonate (pH 10.4) as the mobile phase at 0.2 mL min−1. Samples were diluted in water, filtered (10 kDa MWCO, modified PES; VWR, Darmstadt, Germany) and 10 μL of the samples was applied on the column. Data was analysed with Dionex Chromelion software.
Tcc 3000rs
Manufactured by Thermo Fisher Scientific
Sourced in Germany, United States
The TCC-3000RS is a temperature control and cooling device for laboratory applications. It is designed to precisely control the temperature of samples or equipment within a specified range. The device features digital temperature display and intuitive controls for easy operation.
Automatically generated - may contain errors
Lab products found in correlation
Wps 3000trs, by Thermo Fisher Scientific (10 mentions)
Srd 3600, by Thermo Fisher Scientific (7 mentions)
Hpg 3400rs, by Thermo Fisher Scientific (7 mentions)
Q exactive plus, by Thermo Fisher Scientific (5 mentions)
Dad 3000rs, by Thermo Fisher Scientific (3 mentions)
Ultimate 3000 hplc system, by Thermo Fisher Scientific (2 mentions)
Wps 3000sl, by Thermo Fisher Scientific (2 mentions)
Hypersil gold c18 column, by Thermo Fisher Scientific (2 mentions)
Q exactive plus mass spectrometer, by Thermo Fisher Scientific (2 mentions)
Hpg 3200rs, by Thermo Fisher Scientific (2 mentions)
Dionex ultimate 3000 rslc, by Thermo Fisher Scientific (2 mentions)
Diaion hp 20, by Merck Group (2 mentions)
Chromelion software, by Thermo Fisher Scientific (1 mentions)
Metrosep a supp10 250 40 column, by Metrohm (1 mentions)
Lpg 3400sd, by Thermo Fisher Scientific (1 mentions)
Dad 3000, by Thermo Fisher Scientific (1 mentions)
Chromeleon 7, by Thermo Fisher Scientific (1 mentions)
Dionex ultimate 3000 biors, by Thermo Fisher Scientific (1 mentions)
Accucore c18 column, by Phenomenex (1 mentions)
Wps 3000tbrs, by Thermo Fisher Scientific (1 mentions)
16 protocols using tcc 3000rs
1
HPLC Separation of Glucarate Metabolites
2
HPLC Analysis of Bioactive Compounds in NSTC
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The major components of NSTC are flavonoids, glycosides, phenols, ketosteroids and so on.[8–10 ] Prior to treating the experimental rats, NSTC sample used in this study was analysed to ensure the quality control, with an Ultimate 3000 HPLC system (Dionex, S/N: 8043731 USA) consisting of DGP-3600SD pump, SRD-3600 degasser, WPS-3000SL autosampler, TCC-3000RS column compartment and DAD-3000 diode array detector. The original data were calculated and processed with Chromeleon 6.8 Software. An Ultimate AQ-C18 (150 × 4.6 mm, 3 μm) column was used for the analysis, and acetonitrile (A)–tetrahydrofuran (B)–0.05% phosphoric acid (C) were served as the mobile phase with linear gradient elution in 70 min (A: 2%→20%, B: 0%→10%, C: 98%→70%). The flow rate was 1.1 ml/min with ultraviolet absorbance detection at 254 nm. The operation was carried out at 20 °C. Paeoniflorin, ecdysterone, typhaneoside and isorhamnetin-3-O-neohesperidoside were identified and quantified by the comparison of retention time and ultraviolet spectra with those of standard mix, as shown in Figure 1 .
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High performance liquid chromatography (HPLC) of standard mix (
3
UHPLC Analysis of Synthesized Compounds
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Chromatographic experiments were performed on an Ultimate 3000 UHPLC setup (Thermo Fisher Scientific, Waltham, MA, USA) including a solvent rack (SR-3000), a quaternary pump (LPG-3400SD), an autosampler (WPS-3000SL), a column thermostat (TCC-3000RS) and a diode-array detector (DAD-3000). Analysis was performed under a constant temperature of 25 °C and a flow rate of 0.5 mL min−1 using a Reprosil Saphir column (300 × 8 mm) with 10 μm unmodified silica particles and a mean pore size of 100 nm. An aqueous solution of 2 mM SDS and 8 mM ammonium acetate solution was used as mobile phase. 30 μL of all samples were injected after the synthesis without any further purification. Chromatograms were measured at a wavelength of 450 nm.
4
Paracetamol Quantification in SODF
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For each formulation, three individual SODFs were dissolved in 100 mL of distilled water. Samples of the solutions were then diluted and the drug concentration was determined by ultra-high performance liquid chromatography (UHPLC, Thermofisher Scientific, Waltham, MA, USA) using a UHPLC-DAD system. It consisted of a Thermo Scientific™ Dionex™ UltiMate™ 3000 BioRS equipped with a WPS-3000TBRS autosampler and a TCC-3000RS column compartment set at 35 °C. The system was operated using Chromeleon 7 software (Thermofisher Scientific, Waltham, MA, USA). An Accucore C18 column (2.6 µm, 100 × 2.1 mm2) combined with a security guard ultra-cartridge (Phenomenex Inc., Torrance, CA, USA) was used. An isocratic binary solvent system was utilized, consisting of water/formic acid (1%, v/v) as solvent A and acetonitrile/formic acid (1%, v/v) as solvent B (90%A, 10%B). The flow rate of the mobile phase was 1.5 mL/minute, and the injection volume was 50 μL. Quantitative analysis of paracetamol in the SODFs was carried out using an external standard method. The calibration curve was constructed using 5 different standard levels in the concentration range 1–20 mg/L. The peak of paracetamol was monitored at 244 nm.
5
Dionex UltiMate 3000 LC-MS Protocol
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Chromatographic separation was carried out on a Dionex UltiMate 3000 system (Thermo Fisher Scientific, Waltham, MA) configured with a rapid separation (RS) pump (LPG-3400 RS), an autosampler (WPS-3000 TRS) and a column compartment (TCC-3000 RS), all operated by Chromeleon 6.8 software. Samples were injected on a Hypersil GOLD C18 column (2.1 × 100 mm; Thermo Scientific, Les Ulis, France). Parallel reaction monitoring (PRM and accurate mass measurements were performed on a Q-Exactive Plus mass spectrometer (Thermo Fisher Scientific, Bremen, Germany) with a heated electrospray ionization (H-ESI II) probe. Thermo Xcalibur 3.0.63 software was used for both instrument setup and LC-MS system control during data acquisition, and for data treatment. Q Exactive Plus Tune 2.5 software was used for direct control of the mass spectrometer.
6
HPLC Analysis of Stilbenes
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The analysis of the stilbenes was performed according to the method of Ewald et al. [38 (link)] using high-performance liquid chromatography (UltiMate 3000 HPLC, Thermo scientific, Munich, Germany) that included an HPG-3200RS binary pump, WPS-3000RS/TRS autosampler, TCC-3000RS column oven, and a DAD-3000RS photodiode array detector. HPLC separation was achieved on AcclaimTM 120 C18 columns (120 Ǻ 3 × 150 mm, 3 μm) maintained at 25 °C using a mobile phase consisting of 1% aqueous acetic acid (v/v) (A) and methanol (B). The separation was carried out at 25 °C with a flow rate of 0.6 mL min−1 under the following conditions: 0 min (20% B), 10 min (20% B) 6.5 min (37% B), 12.6 min (50% B), and 21.0 min (100% B). Under these conditions, Rsv and Vf were eluted with a retention time of 14.7 min and 17.8 min and monitored at 306 and 324 nm, respectively. Calibration curves were prepared using the endotoxin standards (Sigma-Aldrich, Steinheim, Germany) of Rsv (R2 = 0.9993) and Vf (R2 = 0.9994) in the concentration range 1–500 mg L−1. The amount of Rsv and Vf found in each extract was expressed as mg of compound kg−1 of DW. Each sample was analysed in duplicate.
7
Quantitative LC-MS/MS Bioanalysis Protocol
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A TSQ Quantum Ultra mass spectrometric detector with electrospray ionization (ESI) interface (Thermo Scientific, USA) was coupled with a Dionex Ultimate 3000 ultra-performance liquid chromatography system consisting of an LPG-3400SDN pump, a WPS-3000TSL autosampler, and a TCC-3000RS column compartment. The separation of analytes and IS was achieved by using a Hypersil GOLD C18 column (50 mm × 2.1 mm, 3 µm; Thermo Scientific, USA) that was maintained at 25°C. The water (containing 0.1% formic acid and solvent A) and acetonitrile (solvent B) was used as mobile phase with gradient elution. The gradient was as follows: 0 min 5% B, 0.5 min 5% B, 2.0 min 80% B, 2.1 min 5% B, and 3.0 min 5% B, and then stopped. The flow rate was set at 0.4 mL/min. The select reaction monitoring (SRM) conditions (source parameters) were defined as follows: spray voltage 3.5 kV, vaporizer temperature 250°C, and capillary temperature 300°C. The optimized mass spectrometric parameters and mass spectra for all the analytes are shown in Table 1 and Figure 1 , respectively. All the operations, acquiring and analysis of data, were controlled using Xcalibur software (Thermo Scientific, USA).
8
HPLC-MS/MS Method for Compound Analysis
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Analyses were performed on a Thermo Scientific (Waltham, MA, USA) system Transcend TLX equipped with binary pump HPG-3200 RS, column thermostate TCC-3000RS, valve interface module and an open auto-sampler PAL-LX1. The chromatographic system was coupled with a TSQ Quantum Access Max triplequadrupol mass spectrometer, with HESI ionization source. Chromatographic separation was performed on a Zorbax Eclipse Plus C18 column (50 mm length, 4.6 mm internal diameter and 1.8 μM particle size). The mobile phase and mass spectrometer conditions were the same with slight modifications in terms of optimization as those from the work of Sobsey et al. [17 (link)].
9
HPLC-Orbitrap MS Metabolomic Analysis
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Standard compounds: LC-MS-grade acetonitrile (ACN), formic acid and methanol (MeOH) were obtained from Fisher Scientific (Fair Lawn, NJ, USA). LC-MS-grade water (18 MX) was prepared using a Millipore Milli-Q purification system (Millipore Corporation, Bedford, MA, USA).
High-performance liquid chromatography (HPLC) separation was performed using a Thermo ScientificTM Dionex Ultimate 3000 LC system (Thermo Fisher Scientific, Waltham, MA, USA) with a Thermo Hypersil Gold aQ (1.9 µm, 100 mm × 2.1 diameter). A Thermo Scientific Q Exactive Focus (Thermo Fisher Scientific, USA) equipped with a pump: HGP-3200RS, Autosampler: WPS3000TRS, column compartment: TCC3000RS, a degasser: SRD3400, Diode Array Detector (DAD), Orbitrap mass analyser with a heated-electrospray ionization (H-ESI II), and software of Xcalibur and Chromaleon was used for LC-MS and LC-MS/MS detection.
High-performance liquid chromatography (HPLC) separation was performed using a Thermo ScientificTM Dionex Ultimate 3000 LC system (Thermo Fisher Scientific, Waltham, MA, USA) with a Thermo Hypersil Gold aQ (1.9 µm, 100 mm × 2.1 diameter). A Thermo Scientific Q Exactive Focus (Thermo Fisher Scientific, USA) equipped with a pump: HGP-3200RS, Autosampler: WPS3000TRS, column compartment: TCC3000RS, a degasser: SRD3400, Diode Array Detector (DAD), Orbitrap mass analyser with a heated-electrospray ionization (H-ESI II), and software of Xcalibur and Chromaleon was used for LC-MS and LC-MS/MS detection.
10
UHPLC-HRMS Analysis of Complex Samples
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UHPLC-HRMS analysis was performed using a Thermo Scientific Dionex Ultimate 3000 RSLC instrument (ThermoFisher Scientific, Germering, Germany) consisting of a six-channel degasser SRD-3600, a high-pressure gradient pump HPG-3400RS, an autosampler WPS-3000TRS and a column compartment TCC-3000RS coupled to a Thermo Scientific Q Exactive Plus mass spectrometer (ThermoFisher Scientific, Inc., Bremen, Germany) . UHPLC separations were performed on a Kinetex С18 column (Phenomenex Corporation, Torrence, CA, USA) (2.1 x 100 mm, 2.6 µm) at 40 °C. Each chromatographic run was carried out with a binary mobile phase consisting of water containing 0.1% (v/v) formic acid (A) and acetonitrile also with 0.1% (v/v) formic acid (B). A gradient program was performed as follows: 0-3 min, 10% B; 3-23 min, 10-70% B; 23-25 min, 95% B. The flow rate was 0.3 mL/min, and the sample injection volume was 1 μL. Full scan data were recorded in (+) ESI mode from m/z 250 to 1 000 at a resolution of 70 000 (at m/z 200). Full scan dd-MS/MS (top 5) was performed at a resolution of 17 500 (at m/z 200), AGS target 1e5 with maximum IT 50 ms. Xcalibur software ver. 4.0 was used for data acquisition and processing.
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