Tissue samples of muscle are extracted by Metware according to standard procedures. The sample extracts were analyzed using an LC-ESI-MS/MS system (UPLC, ExionLC AD, https://sciex.com.cn/ ; MS, QTRAP® System, https://sciex.com/ ). LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (QTRAP), QTRAP® LC-MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by Analyst 1.6.3 software (Sciex). Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period. Significantly regulated metabolites between groups were determined by variable importance in projection (VIP)≥1 and absolute Log2FC (fold change)≥1. VIP values were extracted from OPLS-DA result, which also contain score plots and permutation plots, was generated using R package MetaboAnalystR. The data was log transform (log2) and mean centering before OPLS-DA. In order to avoid overfitting, a permutation test (200 permutations) was performed.
Esi turbo ion spray interface
Manufactured by AB Sciex
Sourced in United States
The ESI Turbo Ion-Spray interface is a key component in mass spectrometry systems manufactured by AB Sciex. Its core function is to efficiently ionize and transfer analyte molecules from the liquid phase into the gas phase, enabling their subsequent detection and analysis by the mass spectrometer.
Automatically generated - may contain errors
Lab products found in correlation
Analyst 1, by AB Sciex (6 mentions)
Qtrap lc ms ms system, by AB Sciex (1 mentions)
Exionlc ad, by AB Sciex (1 mentions)
Qtrap system, by AB Sciex (1 mentions)
Qtrap, by AB Sciex (1 mentions)
1290 infinity hplc system, by Agilent Technologies (1 mentions)
Scientz 100f, by Ningbo Scientz Biotechnology (1 mentions)
Ab4500 q trap uplc ms ms system, by AB Sciex (1 mentions)
7 protocols using esi turbo ion spray interface
1
Metabolomics Analysis of Muscle Tissue
2
Optimized Triple Quadrupole Mass Spectrometry
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The LIT and triple quadrupole (QQQ) scans were obtained using a triple quadrupole-linear ion trap mass spectrometer (Q TRAP) (Sciex, Framingham, MA, USA), API 6500 Q TRAP LC/MS/MS System (Sciex), equipped with an ESI Turbo Ion-Spray interface (Sciex), operating in a positive ion mode and controlled using the analyst 1.6.3 software (Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature, 500 °C; ion spray voltage, (IS) 5500 V; ion source gas I (GSI), gas II (GSII), and curtain gas (CUR) were set at 55, 60, and 25.0 psi, respectively; the collision gas (CAD) was high. Instrument tuning and mass calibration were performed with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes. QQQ scans were acquired as multiple reaction monitoring (MRM) experiments with collision gas (nitrogen) set at 5 psi. The declustering potential (DP) and collision energy (CE) for individual MRM transitions were done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within the period.
3
Hormone Profiling of Root Samples
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Hormone profiling was conducted as described in Ziegler et al. [45 (link)] (for further information see Appendix D ). Root material was homogenized, extracted in methanol, and processed firstly using a hydrophobic solid phase extraction cartridge (Chromabond Sorbent HR-XC, Macherey-Nagel, Düren, Germany) and secondly with an anion exchange solid phase extraction cartridge (Diethylaminoethyl Sephadex (DEAE-Sephadex)). For the root exudates the anion exchange step was omitted.
Analytes were separated by an Agilent 1290 Infinity HPLC system and detected on-line by ESI-MS/MS using an API 3200 triple-quadrupole LC-MS/MS system equipped with an ESI Turbo Ion Spray interface (AB Sciex, Darmstadt, Germany). Triple quadrupole scans were acquired in the multiple reaction monitoring mode (MRM) with Q1 and Q3 set at unit resolution. Scheduled MRM was performed with a window of 90 s and a target scan time of 0.1 s. Selected MRM transitions and compound specific parameters can be found in Ziegler et al. [45 (link)].
Analytes were separated by an Agilent 1290 Infinity HPLC system and detected on-line by ESI-MS/MS using an API 3200 triple-quadrupole LC-MS/MS system equipped with an ESI Turbo Ion Spray interface (AB Sciex, Darmstadt, Germany). Triple quadrupole scans were acquired in the multiple reaction monitoring mode (MRM) with Q1 and Q3 set at unit resolution. Scheduled MRM was performed with a window of 90 s and a target scan time of 0.1 s. Selected MRM transitions and compound specific parameters can be found in Ziegler et al. [45 (link)].
4
Quantitative Metabolite Analysis by LC-MS/MS
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For metabolite detection, the AB4500 Q TRAP LC/MS/MS system with linear ion (LIT) and triple quadrupole (QQQ) sensors was used. The system was fitted with an ESI Turbo Ion-Spray interface, which was run in both the positive and negative ion modes and controlled by Analyst 1.6.3 software (AB Sciex). The ESI source operation parameters were as follows: ion source, turbo spray; source temperature, 550 ℃; ion spray voltage (IS), 5500 V (positive ion mode)/-4500 V (negative ion mode); gas I, gas II and curtain gas (CUR), 50, 60, and 25.0 psi, respectively; and collision-activated dissociation (CAD), high. In the QQQ and LIT modes, instrument tuning and mass calibration were conducted using 10 and 100 μmol/L polypropylene glycol solutions, respectively. QQQ scans were acquired as MRM experiments with the collision gas (nitrogen) set to medium. Through further declustering potential (DP) and collision energy (CE) optimization, the metamorphic potential (DP) and collision energy (CE) of a single MRM transition were determined. Depending on the metabolites eluted in each period, a specific set of MRM transitions was monitored [8 (link)].
5
Targeted Metabolomics Analysis by Q TRAP
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Linear ion trap (LIT) and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (Q TRAP), AB4500 Q TRAP UPLC/MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by the Analyst 1.6.3 software (AB Sciex, Framingham, MA, USA). The ESI source operation parameters were selected according to previous reports [27 (link)]. A specific set of multiple reaction monitoring (MRM) transitions were monitored for each period according to the metabolites eluted within this period.
6
Freeze-Drying and Mass Spectrometry Analysis
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Biological samples are freeze-dried by vacuum freeze-dryer (Scientz-100 F).The freeze-dried sample waLIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (Q TRAP), AB4500 Q TRAP UPLC/MS/MS System, equipped with an ESI Turbo Ion-Spray interface, operating in positive and negative ion mode and controlled by Analyst 1.6.3 software (AB Sciex). QQQ scans were acquired as MRM experiments with collision gas (nitrogen) set to medium. DP and CE for individual MRM transitions was done with further DP and CE optimization. A specific set of MRM transitions were monitored for each period according to the metabolites eluted within this period.
7
Targeted Metabolite Analysis by Q TRAP
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LIT and triple quadrupole (QQQ) scans were acquired on a triple quadrupole-linear ion trap mass spectrometer (Q TRAP), AB4500 QTRAP® System (AB Sciex™, Framingham, MA 01,701, USA). The system was equipped with an ESI Turbo Ion-Spray interface that operated in positive and negative ion mode and was controlled by Analyst 1.6.3 software (AB Sciex™, Framingham, MA 01,701, USA). The ESI source operation parameters were fixed as follows: ions source, turbo spray; source temperature 550 °C; ion spray voltage (IS) 5500 V (positive ion mode)/-4500 V (negative ion mode). The ion source gas I (GSI), gas II(GSII), and curtain gas (CUR) were set at 50, 60, and 25.0 psi, respectively. The collision-activated dissociation (CAD) was high. Instrument tuning and mass calibration were carried with 10 and 100 μmol/L polypropylene glycol solutions in QQQ and LIT modes, respectively. QQQ scans were acquired as MRM (multiple reaction monitoring) experiments with collision gas (nitrogen) set to medium. DP and CE for individual MRM transitions were carried with further DP and CE optimization. A specific set of MRM transitions was monitored for each period according to the metabolites eluted within the target period.
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