Lyophilized EtOAc fractions were dissolved in 50% methanol and chromatographed using an HPLC-MS system. The chromatography apparatus was an Agilent 1200 from Agilent Technologies (Santa Clara, CA). The EtOAc fractions were analyzed at 25 °C with a 250 × 4 mm i.d., 5 μm, Prontosil 120-5-C18-AQ reverse phase column, Bischoff (Leonberg, Germany). Water, 0.1% HCOOH (solvent A) and acetonitrile 0.1% HCOOH (solvent B) were used as mobile phases. The gradient elution program was as follows (v/v): 0 min 1% B, 0.4 min 1% B, 2 min 10% B, 6 min 35% B, 7 min 50% B, 8.8 min 70% B, 10.8 min 92% B, 11 min 100% B and 12 min 100% B, followed by 10 min for re-equilibration. The optimum values of the ESI-MS parameters were: capillary voltage – 4.7 kv; drying gaz temperature 350 °C; drying gas flow 10 L/min; nebulizing gas pressure 35 psi. LC/MS accurate mass spectra were recorded across the range 150–2000 m/z. The detection wavelengths were set at 280 and 360 nm. LC-ESI-MS analyses were carried out in the negative ion mode. This HPLC was coupled to an Esquire 3000 + ion trap mass spectrometer using an ESI source from Bruker Daltonics (Billerica, MA).
Esquire 3000 ion trap mass spectrometer
Manufactured by Bruker
Sourced in United States, Germany
The Esquire 3000+ is an ion trap mass spectrometer manufactured by Bruker. It is designed to perform high-sensitivity mass analysis of a wide range of analytes. The instrument utilizes an ion trap to capture and analyze ions, providing precise mass measurements and detailed structural information.
Automatically generated - may contain errors
Lab products found in correlation
Agilent 1200, by Agilent Technologies (2 mentions)
Luna c18 column, by Phenomenex (2 mentions)
2501 hplc system, by Knauer (1 mentions)
Agilent 1100 hplc instrument, by Agilent Technologies (1 mentions)
Chromafil rc 45 15 ms, by Macherey-Nagel (1 mentions)
Acquity uplc h class system, by Waters Corporation (1 mentions)
Agilent 1100 series hplc system, by Agilent Technologies (1 mentions)
Minispin plus, by Eppendorf (1 mentions)
L tryptophan, by Merck Group (1 mentions)
Sodium chloride (nacl), by Merck Group (1 mentions)
10 protocols using esquire 3000 ion trap mass spectrometer
1
HPLC-MS Analysis of EtOAc Fractions
2
Lipidomic Analysis of Lung and Brain Eicosanoids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Eicosanoids and their derivatives were extracted from the lung and brain, and lipids from the lung were analyzed as previously described by high-pressure liquid chromatography-tandem mass spectrometry (LC-MS/MS) [56 (link)]. Accordingly, for the liquid chromatography, the 1100 Series capillary LC unit (Agilent Technologies Deutschland, Waldbronn, Germany) was used for samples of the lung and brain. Subsequently, the Esquire 3000+ ion trap mass spectrometer (Bruker Corporation, Billerica, MA, USA) was used for lung samples and the Daltonik amaZon SL (Bruker Corporation, Billerica, MA, USA) was used to examine the samples of the brain (settings: Supplementary Table S7 ). The characteristic mass spectrometric extracted ion chromatograms (EIC) traces were used for qualification and quantification (Supplementary Table S8 ).
3
Cyclic NGR Peptide-Daunorubicin Conjugates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cyclic NGR peptide-daunorubicin conjugates were prepared by a combination of solid phase peptide synthesis and chemoselective ligation (oxime bond formation) in solution as described in Tripodi et al. [57 (link)]. The crude peptides and conjugates were purified on a KNAUER 2501 HPLC system (KNAUER, Bad Homburg, Germany) was applied with a semi-preparative Phenomenex Luna C18 column (250 mm × 21.2 mm) with 10 μm silica (100 Å pore size) (Torrance CA). Linear gradient elution (0 min 5% B; 60 min 90% B) with eluent A (0.1% TFA in water) and eluent B (0.1% TFA in MeCN-H2O (80:20, v/v)) was used at a flow rate of 4 mL/min. The resulting fractions were lyophilized. Electrospray Ionization (ESI)-mass spectrometric analyses were carried out on an Esquire 3000+ ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany). The freeze-dried bioconjugates were directly used for the in vitro and in vivo studies.
4
Cyclic NGR Peptide Synthesis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The synthesis of cyclic NGR peptides were described previously (Enyedi et al., 2015) (link). The c[KNGRE]-NH 2 was prepared from semi-protected derivative using BOP coupling agent under the appropriate condition followed by the removal of ClZ protecting group from the side chain of Lys with anhydrous HF. The thioether bond was established in 0.1 M Tris buffer (pH=8.1). The linear precursor peptide was added to the solution in portions for 1 h.
The final peptide concentration was 10 mg/mL in both cases. The reaction mixtures were stirred for further 2 h. The final products were purified (see above) and their purity and identity were analysed. Analytical RP-HPLC was accomplished on the same equipment using Phenomenex Luna C18 column (250 mm x 4.6 mm) with 5 μm silica (100 Å pore size) as a stationary phase with linear gradient elution (0 min 0% B; 5 min 0% B; 50 min 90% B) using the same eluents (above mentioned earlier). The flow rate was 1 mL/min. Peaks were detected at 214 nm. The purity of all compounds was over 95%. The pure products were characterized with ESI-MS by means an equipment of Esquire 3000+ ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany).
The final peptide concentration was 10 mg/mL in both cases. The reaction mixtures were stirred for further 2 h. The final products were purified (see above) and their purity and identity were analysed. Analytical RP-HPLC was accomplished on the same equipment using Phenomenex Luna C18 column (250 mm x 4.6 mm) with 5 μm silica (100 Å pore size) as a stationary phase with linear gradient elution (0 min 0% B; 5 min 0% B; 50 min 90% B) using the same eluents (above mentioned earlier). The flow rate was 1 mL/min. Peaks were detected at 214 nm. The purity of all compounds was over 95%. The pure products were characterized with ESI-MS by means an equipment of Esquire 3000+ ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany).
5
Metabolite Profiling of E. planum Roots
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Identification of secondary metabolites present in the roots of E. planum was performed using the HPLC-MS system consisting of Agilent 1100 HPLC instrument with a photodiode-array detector PDAeλ (Palo Alto, CA, USA) and Esquire 3000 ion trap mass spectrometer (Bruker Daltonics, Bremen, Germany) with the XBridge C18 column (150 × 2.1 mm, 3.5 μm particle size) and the MSn spectra were recorded in the negative and positive ion modes using a previously published approach [28 (link), 29 (link)]. The elution was conducted with water containing 0.1% formic acid (solvent A) and acetonitrile (solvent B). The gradient elution was started at 10% of B and linearly changed to 25% of B in 25 min and to 98% in 46 min of B over 10 min, followed by return to stationary conditions and reequilibration for 10 min. Moreover, the UHPLC-MS/MS was applied for the quantitative analysis of phenolic compounds (flavonoids and phenolic acids) and for the analysis of triterpenoid saponins. Methods and results were described previously [30 (link)].
6
Glutamic Acid Cluster Analysis by ESI-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mass spectra were acquired with an Esquire 3000+ Ion-Trap Mass Spectrometer (Bruker Daltonik) in positive and negative ion modes. Samples were introduced by a syringe pump at a flow rate of 5 μl s−1. Spectra were recorded by scanning from 120 to 2,500 m/z. The ion source parameters were: 10 psi nebulizer gas (nitrogen), 6 l min−1 drying gas (nitrogen) with a temperature of 200 °C and capillary voltage 3,500 V. The existence and stability of glutamic acid clusters were probed by several series of measurements with Glu solutions at different concentrations (0.1–50 mM), different pH values (native and pH 11), in the presence and absence of an internal standard (aspartic acid), with and without co-solvent (methanol) and after different ageing times. Generally, very similar observations were made under all investigated conditions, that is, large oligomers were detected in all samples. There was no effect of ageing time on the results of the ESI-MS measurements.
7
HPLC-DAD-ESI-MS Analysis of EtOAc Fractions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Lyophilised EtOAc fractions were dissolved in 50% methanol and chromatographed using HPLC-DAD-ESI-MS system. The chromatography apparatus was Agilent 1200 from Agilent Technologies (Santa Clara, CA, USA). The EtOAc fractions were analyzed at 25°C with a 250 × 4 mm i.d. 5 μm, Prontosil 120-5-C18-AQ reverse phase column (Bischoff, Leonberg, Germany). Water, 0.1% HCOOH (solvent A), and acetonitrile 0.1% HCOOH (solvent B) were used as mobile phases. The gradient elution program was as follows (v/v): 0 min 1% B, 0.4 min 1% B, 2 min 10% B, 6 min 35% B, 7 min 50% B, 8.8 min 70% B, 10.8 min 92% B, 11 min 100% B, and 12 min 100% B, followed by 10 min for reequilibration. The optimum values of the ESI-MS parameters were as follows: capillary voltage, −4.7 kv; drying gas temperature, 350°C; drying gas flow, 10 L/min; nebulising gas pressure, 35 psi. LC/MS accurate mass spectra were recorded across the range 150–2000 m/z. The detection wavelengths were set at 280 and 360 nm. LC-ESI-MS analyses were carried out in the negative ion mode. This HPLC was coupled to Esquire 3000+ ion trap mass spectrometer using ESI source from Bruker Daltonics (Billerica, MA, USA).
8
HPLC-based Quantification of Pineapple Juice Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For HPLC analysis, pineapple juices were centrifuged (10,000×g, 10 min, miniSpin plus, Eppendorf, Hamburg, Germany) and the obtained supernatants membrane-filtered (0.45 μm, regenerated cellulose, Chromafil RC-45/15 MS, Macherey-Nagel, Düren, Germany) into amber glass vials.
For compound identification, HPLC-DAD-ESI-MS n analyses were conducted using an Agilent series 1100 HPLC system (Agilent Technologies, Waldbronn, Germany) coupled to an Esquire 3000+ ion-trap mass spectrometer with electrospray ionisation (ESI) source (Bruker Daltonics, Bremen, Germany) as reported by Steingass et al. (2015) (link).
Quantitation was achieved applying an Acquity H-class UPLC system equipped with an eλ photodiode array detector (both from Waters, Milford, MA, USA). HPLC settings were as previously reported (Difonzo et al. 2019) (link). Detection wavelengths were set to 260, 280, and 320 nm. UV/Vis spectra were recorded in the range of 200-700 nm. Quantitation was achieved using external calibrations of suitable reference standards as detailed elsewhere (Difonzo et al. 2019) (link).
For compound identification, HPLC-DAD-ESI-MS n analyses were conducted using an Agilent series 1100 HPLC system (Agilent Technologies, Waldbronn, Germany) coupled to an Esquire 3000+ ion-trap mass spectrometer with electrospray ionisation (ESI) source (Bruker Daltonics, Bremen, Germany) as reported by Steingass et al. (2015) (link).
Quantitation was achieved applying an Acquity H-class UPLC system equipped with an eλ photodiode array detector (both from Waters, Milford, MA, USA). HPLC settings were as previously reported (Difonzo et al. 2019) (link). Detection wavelengths were set to 260, 280, and 320 nm. UV/Vis spectra were recorded in the range of 200-700 nm. Quantitation was achieved using external calibrations of suitable reference standards as detailed elsewhere (Difonzo et al. 2019) (link).
9
HPLC-MS/MS Analysis of Peptides
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HPLC-MS/MS analyses and data processing was performed according to Santos-Hernández et al. 28 Prior to the analyses, the reduction step was performed with 146 mM DTT. Gradient elution was performed with and Agilent 110 HPLC separation system (Agilent, CA, USA). Detection was performed with a Bruker Daltonics Esquire 3000 Ion Trap mass spectrometer (Bruker, MA; USA) in a full scan acquisition mode from 100 to 2300 m/z, and setting target mass at 450, 750 and 1200. Biotools version 3.2 was used for interpreting the matched MS/ MS spectra. For peptide profile visualization, the web application Peptigram was used. 29
10
Infrared Spectroscopy of Tryptophan Clusters
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
) were recorded at the Centre Laser Infrarouge d'Orsay (CLIO) free electron laser (FEL) facility at the University of Paris XI. 37, 38 Details of the experimental apparatus have been previously described. 12, 39, 40 Briefly, monomeric and dimeric Trp clusters were produced by electrospray ionization (ESI) in negative mode of ca. 100 μM solutions of L-tryptophan in methanol/water (50/50 vol%) containing ca. 5 μM of NaCl. Ltryptophan and NaCl were obtained from Sigma Aldrich (Oakville, ON, Canada) and used without further purification. The nascent clusters produced were transferred to a Bruker Esquire 3000+ ion trap mass spectrometer, where they were mass selected and subsequently irradiated by the tunable output of the FEL. IRMPD spectra were acquired by irradiating the trapped ions for 400 ms at 5 cm -1 intervals in the 850 -1900 cm -1 range. The IR-FEL output was delivered to the trapped ions as a series of 9 ms macropulses emitted with a repetition rate of 25 Hz. Each macropulse consisted of 600 picosecond micropulses. The average IR power for these experiments varied approximately quadratically as a function of photon wavenumber between 800-1200 mW, with the peak power being delivered near the midpoint of the spectrum. Vibrational spectra were generated by recording the fragmentation efficiency of the Trp clusters as a function of FEL wavenumber.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!