The IOP concentration in the influent and effluent of both bioreactors was determined by high-performance liquid chromatography (HPLC) using an Agilent Technology 1260 series chromatograph as previously described (Toral-Sánchez et al. 2016 ; Toral-Sánchez et al. 2017 (link)). Identification of biotransformation byproducts of IOP was carried out by HPLC coupled to mass spectroscopy (HPLC-MS) in a Varian ® 500-MS ion trap mass spectrometer as reported in previous work (Toral-Sánchez et al. 2017 (link)). VSS and COD was estimated according to standard methods (Eaton and Franson 2005 ). The ORP and pH of the biological system was monitored using a Thermo Scientific STARA2110 Orion Star A211 benchtop pH meter with electrode stand.
500 ms ion trap mass spectrometer
Manufactured by Agilent Technologies
Sourced in United States
The 500-MS ion trap mass spectrometer is a laboratory instrument designed for high-performance mass analysis. It utilizes ion trap technology to capture, store, and analyze ions, enabling the identification and quantification of chemical compounds. The core function of the 500-MS is to provide accurate mass measurements and detailed structural information about samples.
Automatically generated - may contain errors
Lab products found in correlation
Pursuit xrs c18 column, by Agilent Technologies (2 mentions)
1260 series chromatograph, by Agilent Technologies (1 mentions)
Pierce c18 tip, by Thermo Fisher Scientific (1 mentions)
Sequencing grade modified trypsin, by Promega (1 mentions)
300 mhz nmr spectrometer, by Agilent Technologies (1 mentions)
Ms workstation software, by Agilent Technologies (1 mentions)
Prostar 410, by Agilent Technologies (1 mentions)
Prostar 335, by Agilent Technologies (1 mentions)
6 protocols using 500 ms ion trap mass spectrometer
1
Quantification of IOP Biodegradation Byproducts
2
Proteolytic Resistance Evaluation of dUSTBβP3
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Proteolytic resistance of dUSTBβP 3 and previously reported dUSCL di-C9-KKKK-NH2 was evaluated with tryptic digest assay as previously described (Ramirez et al., 2020 (link)). The compounds were diluted with 50 mM ammonium bicarbonate (pH 7.8) and were incubated with sequencing-grade modified trypsin from Promega (United States) at a molar ratio of 1:5,000 (enzyme/compound) at 37°C for 2 h. Termination of the reaction was done by overnight freezing at −18°C, and the samples were purified and concentrated by using Pierce C18 tips (10 μL) from Thermo Scientific (United States). Stability toward trypsin was assessed by mass fragmentation analysis in positive ion mode on a Varian 500-MS ion trap mass spectrometer (United States).
3
HPLC and HPLC-MS Analysis of IOP
4
NMR Spectroscopy and Mass Spectrometry Analysis
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The synthesized compounds were structurally elucidated using a Varian 300 MHz NMR spectrometer (Palo Alto, CA, USA) using DMSO-d6 and CDCl3 at 99.9% D. Coupling constants (J) are expressed in hertz (Hz). Chemical shifts (δ) are reported in parts per million (ppm) units relative to the reference (TMS). Mass spectrometry (MS) analysis was performed on a Varian 500 MS ion trap mass spectrometer using electrospray ionization (ESI). NMR and MS spectra of compounds 3 –6 are available in the Supplementary materials .
Melting points are determined on a Gallenkamp MFB-595 melting point apparatus (London, UK) and are uncorrected.
All commercially available starting materials and solvents were used without further purification.
Melting points are determined on a Gallenkamp MFB-595 melting point apparatus (London, UK) and are uncorrected.
All commercially available starting materials and solvents were used without further purification.
5
LC-IT-MS/MS Analysis of Compounds
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Liquid chromatography-ion trap-mass spectrometry/mass spectrometry (LC-IT-MS/MS) analysis was performed using a Varian 500-MS ion trap mass spectrometer (Varian, USA), which consisted of an LC pump (Varian 212), an auto sampler (Prostar 410), and a photodiode array detector (Prostar 335). The LC system was equipped with a Varian PurSuit XRs C18 column (i.d., 100 mm × 2.0 mm; 3 µm particle size; Varian, Lake Forest, CA, USA). The initial mobile phase conditions consisted of 90% A (0.1% (v/v) formic acid in water) and 10% B (0.1% (v/v) formic acid in acetonitrile), which was maintained for 2 min, followed by an increase to 90% B for 25 min. The gradient was maintained at 90% B for 5 min, rapidly decreased to 50% B for 0.06 min, and then maintained for 5 min. The flow rate was set to 0.2 ml/min and the volume of sample injected was 10 µl. The full-scan mass spectral range was 100-1,000 m/z. The operating parameters for analyzing the samples were as follows: spray needle voltage, 5 kV; capillary voltage, 80 V; drying temperature, 300°C; drying gas pressure (nitrogen), 20 psi; nebulizer gas pressure (air), 40 psi. Tandem mass spectrometry analysis was carried out using scan-type turbo data-dependent scanning (DDS) under the same conditions. LC-IT-MS/MS data were analyzed using the MS workstation software (ver. 6.9; Varian, USA).
6
Quantitative Analysis of Metabolites by LC-ESI-MS/MS
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Liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) was performed using a Varian 500-MS ion trap mass spectrometer (Varian, Palo Alto, CA, USA), which consisted of an LC binary pump (Varian 212), a photodiode array detector (Prostar 335), and an autosampler (Prostar 410). The LC system was equipped with a Varian PurSuit XRs C18 column (100 × 2.1 mm i.d. × 3 µm particle size). Mobile phases consisted of water (A) and acetonitrile (B) with 0.1% formic acid (v/v). The initial mobile phase consisted of 90% A and 10% B, which was maintained for 2 min, followed by an increase to 90% B over 25 min, which was maintained at 90% B for 5 min, and then rapidly decreased to 10% B, which was maintained for 5 min. The flow rate was set to 0.2 ml/min and 10 µl of sample was injected. The full-scan mass spectral range was 100-1,000 m/z. The running parameters for analyzing the samples were as follows: spray needle voltage, 5 kV; capillary voltage, 80 V; drying temperature, 300°C; drying gas (nitrogen) pressure, 20 psi; nebulizer gas (air) pressure, 40 psi. Tandem mass spectrometry analysis was carried out using scan-type turbo data-dependent scanning (DDS) under the same conditions. LC-ESI-MS/MS data were analyzed using the MS workstation software (ver. 6.9; Varian, USA).
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