L-Erg and S-Met-L-Erg determination in the urines of the test experiment was carried out as previously described in Ref. [26 (link)]. The analysis of L-Erg and S-Met-L-Erg in the urines of low and high-dose treatment was performed on an Agilent 1290 Infinity II UHPLC system (Agilent Technologies, Santa Clara, USA) coupled to a 6500 QTRAP mass spectrometer equipped with Ion Drive Turbo V ion source LC-MS/MS system (Sciex, Framingham, MA, USA). L-Erg and S-Met-L-Erg concentration was normalized by urinary creatinine, determined with the Creatinine Assay Kit (Sigma-Aldrich, MA, USA).
Ion drive turbo 5
Manufactured by AB Sciex
Sourced in United States, Germany
The Ion Drive Turbo V is a high-performance ion source for mass spectrometry applications. It utilizes a turbomolecular pump to generate a high vacuum, enabling efficient ion generation and transfer into the mass spectrometer.
Automatically generated - may contain errors
3 protocols using ion drive turbo 5
1
Quantification of L-Erg and S-Met-L-Erg in Urine
2
UHPLC-MS/MS Metabolite Analysis
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The supernatant of each sample (MRS, ACS, and BCS) was mixed with an extraction solution. After centrifugation, UHPLC-MS/MS analysis was performed. The UHPLC separation was processed using an EXIONLC System (Sciex), a SCIEX 6500 QTRAP plus triple quadrupole mass spectrometer (Sciex) equipped with an IonDrive Turbo V electrospray ionization interface. SCIEX Analyst Work Station_v1.6.3 and Sciex MultiQuant_v3.0.3 were employed for MRM data acquisition and processing.
3
Simultaneous Analysis of Vitamins and Nucleotides
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MS analysis was performed in the MRM‐scan mode on a Q‐Trap 6500+ (Sciex, Darmstadt, Germany) equipped with an Ion Drive Turbo V electrospray ionization probe. In both the positive and negative ionization modes, the source temperature was set to 550°C, the curtain gas was set to 30, the nebulizer gas (GS1) and the auxiliary gas (GS2) were both set to 45, and the collision activation dissociation (CAD) gas was set to medium. Nitrogen was used as both the curtain gas and the CAD gas. In the positive ionization mode, the spray voltage was set to 5.5 kV, the declustering potential to 90, and the entrance and exit potentials to 10. In the negative ionization mode, the spray voltage was set to −4.5 kV, and the declustering potential to −60 and the entrance and exit potentials were both set to −10. Both the LC and the MS were controlled using Analyst software version 1.7.
The MRM transitions used for analysis of the vitamins were as follows: Folate 442/295.1, Riboflavin 377/243 and FAD 786/348 all analysed in the positive ionization mode, and, for FMN 455/213 analysed in the negative mode. The MRM transitions used for the analysis of the nucleotides were as follows: IMP 349/137, UMP 325/97, GMP 364/154, AMP 348/136 all in the positive ionization mode.
Data analysis was performed using the Sciex OS software. A Gaussian smoothing of 1 point was applied to all peaks.
The MRM transitions used for analysis of the vitamins were as follows: Folate 442/295.1, Riboflavin 377/243 and FAD 786/348 all analysed in the positive ionization mode, and, for FMN 455/213 analysed in the negative mode. The MRM transitions used for the analysis of the nucleotides were as follows: IMP 349/137, UMP 325/97, GMP 364/154, AMP 348/136 all in the positive ionization mode.
Data analysis was performed using the Sciex OS software. A Gaussian smoothing of 1 point was applied to all peaks.
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