HPLC analysis was carried out using an Agilent 1100 series system coupled with a photodiode array detector. UV and FT-IR spectra were obtained employing a Perkin Elmer Lambda25 and a Jasco FT/IR-4200 instrument, respectively. All NMR spectra were recorded on Bruker Avance III 400 and 600 spectrometers. Spectra were referenced to the residual solvent signal of acetone-d6 with resonances at δH/C 2.04/29.8. HR-ESI-TOF-MS data were recorded on a Bruker maXis 4G mass spectrometer. ESI-LC/MS experiments were carried out using an Agilent 1200 series system coupled with an ESI spectrometer (LC/MSD Ultra Trap System XCT 6330).
Ultra trap system xct 6330
Manufactured by Agilent Technologies
Sourced in Germany
The Ultra Trap System XCT 6330 is a laboratory instrument designed for the removal of contaminants from gas samples. It features a trapping system that effectively captures and retains a wide range of impurities, ensuring the purity of the gas sample for downstream analytical processes.
Automatically generated - may contain errors
Lab products found in correlation
1200 series hplc, by Agilent Technologies (2 mentions)
1100 series system, by Agilent Technologies (1 mentions)
1200 series system, by Agilent Technologies (1 mentions)
Ft ir 4200, by Jasco (1 mentions)
Lambda25, by Jasco (1 mentions)
Maxis 4g mass spectrometer, by Bruker (1 mentions)
6300 series trap control version 6, by Bruker (1 mentions)
Nucleosil 100 c18 column, by Dr. Maisch (1 mentions)
4 protocols using ultra trap system xct 6330
1
Comprehensive Analytical Characterization
2
Extraction and LC-MS Analysis of Cystargolides
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For the extraction of cystargolide, 750 μl of cell culture was thoroughly mixed with the same volume of ethyl acetate (EtOAc), followed by separation of the phases by centrifugation at 1500g for 5 min. The organic phase was collected and evaporated to dryness in a vacuum concentrator. The extracts were dissolved in acetonitrile (MeCN) and subjected to LC–MS analysis. The routine analysis of culture extracts by LC–MS and MS/MS was performed on a LC/MSD Ultra Trap System XCT6330 (Agilent 1200 series; Agilent Technologies). Samples (2.5 μl) were injected onto a reverse phase column (Nucleosil 100-3 C18, particle size 3 μm, pore size 100 Å, 100 × 2 mm fitted with a precolumn 10 × 2 mm) at a flow rate of 0.4 ml/min and a linear gradient of solvent B from t0 = 0% to t15 = 100% (solvent A: water with 0.1% formic acid, solvent B: MeCN with 0.06% formic acid) for cystargolide A and B detection. Ionization and mass analysis was performed by ESI (positive and negative ionization) in Ultra Scan mode with a capillary voltage of 3.5 kV and a drying gas temperature of 350 °C.
3
Stability Analysis of 12IGF Peptide
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To confirm the stability of 12IGF under standard assay conditions (i.e., to exclude proteolysis of 12IGF by ClpP), HPLC-MS analysis was performed on an Agilent 1200 HPLC series equipped with a temperature-controlled sampler and coupled to an Ultra Trap System XCT 6330 (Agilent, Waldbronn, Germany). 5 μL of sample (1 μM SaClpP, 46 μM 12IGF in SaClpP activity buffer) were injected at given time points into a Reprosil-Gold 300 C-18 5 μm column (10 × 2 mm precolumn and 100 × 2 mm separation column, Dr. Maisch HPLC GmbH, Ammerbuch, Germany). Separation was conducted with a linear gradient elution over 55 min from 0% B to 100% B at a flow rate of 400 μL/min. Eluents A and B were composed of 0.1% formic acid in water and 0.06% formic acid in methanol, respectively. Absorption spectra were recorded at wavelengths 220 nm, 260 nm, 280 nm, 360 nm, and 435 nm, while 12IGF showed the strongest signal at 220 nm. Electronspray ionization (ESI) was performed in ultra-scan mode (positive and negative, alternating) with a capillary voltage of 3.5 kV and a drying gas temperature of 350 °C. Data analysis was performed with the software 6300 Series Trap Control Version 6.1 (Bruker Corporation, Billerica, MA, USA).
4
Quantification of brasilicardin congeners
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HPLC/MS analysis was performed with an Agilent 1200 HPLC series coupled with an Ultra Trap System XCT 6330 (Agilent, Waldbronn, Germany). Samples (2.5 μl) were injected on a 3 μm Nucleosil 100 C18 column (100 × 2 mm, fitted with a precolumn 10 × 2 mm, Dr. Maisch HPLC GmbH, Ammerbuch, Germany) and separated with eluent A 0.1% formic acid in water and eluent B 0.06% formic acid in acetonitrile by gradient elution (20%–50% B over 10 min followed by 50%–100% B over 5 min) at a flow rate of 400 μl/min. Detection was carried out at 220, 240, 300, 360 and 435 nm. Electrospray ionization in ultra scan mode (positive and negative, alternating) was done with a capillary voltage of 3.5 kV and a drying gas temperature of 350 °C [26] . Detection of m/z values was conducted with Agilent DataAnalysis for 6300 Series Ion Trap LC/MS Version 3.4 (Bruker Daltonik GmbH, Billerica, USA). MS2 and MS3 analyses were performed in positive mode under the same conditions. To determine the concentration of brasilicardin congeners produced by A. japonicum::pPS1, isolated BraC and BraC-agl ranging from 1 to 1000 mg/l in 10 dilution steps were used to generate a standard curve in HPLC/MS. The concentration of brasilicardin congeners was then determined by analysing the peak height of the MS spectrum in regards to the standard curve.
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