Mass spectrometric analysis was performed on an ion trap mass spectrometer equipped with an electrospray ion source ESI (LCQ-DECA, Thermo Fischer Scientific, San Jose, CA, USA). The mass spectrometer was coupled online with a and autosampler (Thermo Fischer Scientific, San Jose, CA, USA) and a LC-pump (Surveyor MS Pump, Thermo Fischer Scientific, San Jose, CA, USA). Samples were dissolved in methanol ((25 µg/µL) and 5µL were loaded onto a Waters Symmetry RP-C18 column (150 mm × 1 mm i.d., 100 Å, 3.5 µm). Separation was achieved thermosetting the column at 25 °C with a linear gradient of H2O + 1% FA and ACN + 1% FA at 50 μL/min. Elution was performed, increasing solvent B from 5% to 15% in 25 min, 25% in 40 min, 30% in 45 min, and 55% in 55 min. Full scan mass spectra were acquired in negative ion mode in the m/z range 150–2000. ESI ion source operated with 220 °C capillary temperature, 30 a.u. sheath gas, −3.5 kV source voltage and −18 V capillary voltage. Mass spectrometric analysis was performed by the data-dependent method with normalized collision energy of 30 a.u. and activation Q set as 0.250. Mass calibration was achieved with a standard mixture of caffeine (Mr 194.1 Da), MRFA peptide (Mr 524.6 Da), and Ultramark (Mr 1621 Da). Data acquisition and data analyses were performed with the Xcalibur v. 1.3 Software.
Symmetry rp c18 column
Manufactured by Waters Corporation
Sourced in United States
The Symmetry RP-C18 column is a chromatography column designed for reverse-phase liquid chromatography (RPLC) applications. It features a C18 stationary phase that provides high-performance separation of a wide range of non-polar and moderately polar analytes.
Automatically generated - may contain errors
Lab products found in correlation
Thermo xcalibur chemstation, by Thermo Fisher Scientific (4 mentions)
Lcq deca, by Thermo Fisher Scientific (1 mentions)
Surveyor ms pump, by Thermo Fisher Scientific (1 mentions)
Tsq quantum access max mass spectrometer, by Thermo Fisher Scientific (1 mentions)
2996 photodiode array detector, by Waters Corporation (1 mentions)
600 hplc pump, by Waters Corporation (1 mentions)
Tsq quantum access max, by Thermo Fisher Scientific (1 mentions)
Thermo accela 600 series hplc system, by Thermo Fisher Scientific (1 mentions)
8 protocols using symmetry rp c18 column
1
Mass Spectrometric Analysis of Compounds
2
HPLC-MS Analysis of WUE-A4 Compounds
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HPLC-UV/ESI-MS/MS was carried out to characterize the components in WUE-A4 before and after ultrafiltration by using a Thermo Accela 600 series HPLC connected with a TSQ Quantum Access MAX mass spectrometer (Thermo Fisher Scientific, San Jose, CA, USA). A Waters Symmetry RP-C18 column (4.6 × 250 mm, 5 µm) was used to perform chromatographic analysis at 30 °C, and the mobile phase consisted of H2O with 0.1% formic acid (A) and ACN (B). The optimized HPLC elution procedures were as follows: 0–15 min, 17% B; 15–40 min, 17–30% B, 40–42 min: 30–56% B. The flow rate was 0.8 mL/min, the injection volume was 10 µL, and the HPLC-UV chromatograms were detected at a wavelength of 254 nm. The negative ion modes were applied to obtained ESI-MS/MS data. Moreover, the parameters of instrument were set as follows: the vaporizer temperature was 350 °C, the capillary temperature was 250 °C, the spray voltage was 3000 V, the cone voltage energy was 40 V, the collision energy was 10 V, the sheath gas pressure was 40 psi, the aux gas pressure was 10 psi, the drying gas flow rate was 6.0 L/min, and the mass range was set from 50 to 1100 (m/z) in the full-scan mode. Finally, the Thermo Xcalibur ChemStation (Thermo Fisher Scientific) was used for data acquisition and analysis.
3
Analysis of Ficus glumosa Metabolites
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The analysis of Ficus glumosa was conducted using a Thermo Accela HPLC 600 connected with a mass spectrometer, TSQ-QuantumTM Access MAX (Thermo Fischer, San Jose, California, USA). The separation of the sample was achieved by Waters Symmetry RP-C18 column, 4.6 × 250 mm, 5 µm (Milford, USA) at 30 °C. The solvents (water/acetonitrile) were composed of formic (FA) acid (0.1%) in ultrapure water (A mobile phase) and 100% ACN (B mobile phase). The HPLC elution gradient was adjusted and set, as follows: 15–17% in 0–5 min., 17% in 5–15 min., 17–23% in 15–40 min., 23–25% in 40–45 min., and 25–33% in 45–65 min. Injected volume, 10 µL, 0.8 mL/min. as the flow rate, and online monitoring of UV-chromatogram was at 280 nm. The MS settings were adjusted, as below: negative full scan and dependent-data scan mode, capillary temperature, 350 °C, vaporizer temperature at 300 °C, sheath gas pressure (N2) at 40 psi, auxiliary pressure (N2) at 10 psi, and spray voltage at 3kV and mass range at 150–1500 m/z.
4
Analytical HPLC Measurements of Samples
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Analytical HPLC measurements were run on a Waters 600 HPLC pump equipped with a Waters 2996 photodiode array detector, a 20 μL Rheodyne injector and a computer‐integrating apparatus. The column was a Waters Symmetry RP‐C18 column (4.6×150 mm, 5 μm). The mobile phase was a mixture of TFA 0.01 M/ACN/(MeOH/THF 90 : 10) 20/52/28 and the flow rate was 0.7 mL/min. The UV detector was set at a length of 254 nm.
5
HPLC-ESI-MS/MS Analysis of Compounds
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The HPLC analysis was performed on a Thermo Access 600 HPLC system (Thermo Fisher Scientific, San Jose, CA, USA). Briefly, a Waters Symmetry RP-C18 column (4.6 mm × 250 mm, 5 µm) with a guard column was served for the chromatographic separations. The mobile phases consisted of 0.1% FA – H2O (A) and ACN (B). The separation was conducted with the optimized elution conditions as follows: 0 – 40 min, 5 – 95% B. The samples (10 µL) were injected into the system with a flow rate of 0.8 mL/min at 30 ℃, and the eluent was monitored at a wavelength of 292 nm.
As for the ESI-MS/MS analysis, the mass spectrometer with electrospray ionization (ESI) was operated to obtain diverse fragment ions in the positive ion mode with the spray voltage of 3.0 kV and the cone voltage of 40.0 V; the vaporizer temperature and capillary temperature were set at 350 ℃ and 250 ℃, respectively; the sheath gas pressure and aux gas pressure were recognized as 40 psi and 10 psi, respectively; the scan range was m/z 150 Da to 1000 Da; the collision energy altered from 30 eV to 45 eV for the MS/MS analysis. All the analytical data was obtained from the professional software of Thermo Xcalibur ChemStation (Thermo Fisher Scientific).
As for the ESI-MS/MS analysis, the mass spectrometer with electrospray ionization (ESI) was operated to obtain diverse fragment ions in the positive ion mode with the spray voltage of 3.0 kV and the cone voltage of 40.0 V; the vaporizer temperature and capillary temperature were set at 350 ℃ and 250 ℃, respectively; the sheath gas pressure and aux gas pressure were recognized as 40 psi and 10 psi, respectively; the scan range was m/z 150 Da to 1000 Da; the collision energy altered from 30 eV to 45 eV for the MS/MS analysis. All the analytical data was obtained from the professional software of Thermo Xcalibur ChemStation (Thermo Fisher Scientific).
6
HPLC-MS/MS Analysis of WE-P Components
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Thermo Accela 600 series HPLC tandem TSQ Quantum Access MAX mass spectrometer (Thermo Fisher Scientific, San Jose, CA, USA) was used to identify the components in WE-P and the ultrafiltration residues [40 (link)]. Briefly, a Waters Symmetry RP-C18 column (4.6 × 250 mm, 5 µm) was used for chromatographic analysis. The mobile phase consisted of 0.1% FA-H2O (A) and ACN (B). The optimized gradient HPLC elution procedures were set as follows: 0–28 min, 30% B; 28–32 min, 30–40% B, 32–60 min: 40–45% B. The injection volume was 10 µL, the flow rate was 0.8 mL/min, the column temperature was 30 °C, and the HPLC-UV chromatograms were obtained at a detection wavelength of 230 nm. The ESI-MS/MS parameters were applied as follows: the negative ion mode was set in this system, the spray voltage was set as 3.0 kV, the cone voltage was set as 40 V, the collision energy was set as 10 V according to the results of fragments; the sheath gas (N2) pressure was set as 40 psi, the aux gas pressure was set as 10 psi, the nebulizing gas flow rate was set as 6.0 L/min, the vaporizer temperature was set as 350 °C, the capillary temperature was set as 250 °C, the mass range from 50 to 1100 (m/z) was set in the full-scan mode. Finally, data acquisition and analysis were carried out by the Thermo Xcalibur ChemStation (Thermo Fisher Scientific, San Jose, CA, USA).
7
HPLC-MS/MS Analysis of Compounds
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The HPLC-UV/ESI-MS/MS analysis was performed using a TSQ Quantum Access MAX mass spectrometer in the negative mode, which was connected with a Thermo Accela 600 series HPLC system (Thermo Fisher Scientific, San Jose, CA, USA). The chromatographic separation of samples was carried out using a Waters Symmetry RP-C18 column (250 mm × 4.6 mm, 5 μm) at 25 °C. The mobile phases consisted of 0.1% (v/v) formic acid–water (A) and acetonitrile (B). The HPLC elution procedures were optimized as follows: 0–30 min, 8–30% B; 30–40 min, 30–95% B. The injection volume of all the samples was 10 μL, the flow rate was set at 800 μL/min, and the wavelength was monitored at 280 nm. Moreover, the optimized parameters of the MS instrument were implemented as follows: capillary temperature of 350 °C; vaporizer temperature of 300 °C; spray voltage of 3,000 V; sheath gas (nitrogen, N2) of 40 psi; auxiliary gas (N2) of 10 psi; and mass range (m/z) of 100–1,500. The mass spectrum data were obtained in the full-scan and the data-dependent mode and analyzed by the Thermo Xcalibur ChemStation (Thermo Fisher Scientific).
8
Kinetics of Pt(IV) Complex Hydrolysis
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150 L of a solution of 1 (0.8 mM) in DMSO was diluted to 3.0 mL with standard physiological saline solution (0.9 %, w/v; pH = 6.2) to obtain a final concentration of 40 M of complex 1 in saline solution. The mixture was incubated at 37 °C in the dark and the hydrolysis reaction was followed by monitoring the decrease of the chromatographic peak of the starting Pt(IV) complex by HPLC-UV. Stationary phase: Waters Symmetry RP-C18 column, 5 M, 4.6 × 250 mm, 100 Å.
Mobile phase: phase A = water and phase B = Acetonitrile; isocratic elution 5% phase B for 5 min, linear gradient from 5% to 20% phase B in 6.5 min, isocratic elution 20% phase B for 5.5 min, linear gradient from 20% to 100% phase B in 10 min, isocratic elution 100% phase B for 5 min, linear gradient from 100% to 5% phase B in 1 min, isocratic elution 5% phase B for 10 min. Flow rate = 0.6 mL min 1 . UV-visible detector set at 220 nm.
Mobile phase: phase A = water and phase B = Acetonitrile; isocratic elution 5% phase B for 5 min, linear gradient from 5% to 20% phase B in 6.5 min, isocratic elution 20% phase B for 5.5 min, linear gradient from 20% to 100% phase B in 10 min, isocratic elution 100% phase B for 5 min, linear gradient from 100% to 5% phase B in 1 min, isocratic elution 5% phase B for 10 min. Flow rate = 0.6 mL min 1 . UV-visible detector set at 220 nm.
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