The purity and identity
of all compounds
was verified using a Shimadzu 2010 LCMS system, consisting of a LC-20AD
binary solvent pumps, a DGU-20A degasser unit, a CTO-20A column oven,
and a SIL-20A HT auto sampler. A Shimadzu SPD-M20A diode array detector
was used for detections. A full spectra range of 190–600 nm
was obtained during analysis. Chromatographic separations were obtained
using a Phenomenex Luna C18 analytical column (5 μm, 50 ×
4.6 mm i.d.) The column was protected by a Phenomenex C18 column guard
(5 μm, 4 × 3.0 mm i.d.). All equipment was controlled and
integrated by Shimadzu LCMS solutions software version 3. Mobile phases
for LCMS analysis were HPLC grade or LCMS grade obtained from Sigma-Aldrich
and Fisher Scientific. The mobile phases consisted of a mixture LCMS
grade Acetonitrile/water (both with 0.1% formic acid for a pH of 2.7).
The initial setting for analysis was set at 5% acetonitrile (v/v),
then was linearly increased to 95% acetonitrile over 6 min. The gradient
was then held at 95% acetonitrile for 2 min and then linearly decreased
to 5% over 0.10 min and held until stop for an additional 1.90 min.
The total run time was equal to 12 min. The total flow rate was set
to 0.5 mL/min. The column oven and flow cell temperature for the diode
array detector was set at 30 °C. The auto sampler temperature
was held at 15 °C. 5uL was injected for analysis.
of all compounds
was verified using a Shimadzu 2010 LCMS system, consisting of a LC-20AD
binary solvent pumps, a DGU-20A degasser unit, a CTO-20A column oven,
and a SIL-20A HT auto sampler. A Shimadzu SPD-M20A diode array detector
was used for detections. A full spectra range of 190–600 nm
was obtained during analysis. Chromatographic separations were obtained
using a Phenomenex Luna C18 analytical column (5 μm, 50 ×
4.6 mm i.d.) The column was protected by a Phenomenex C18 column guard
(5 μm, 4 × 3.0 mm i.d.). All equipment was controlled and
integrated by Shimadzu LCMS solutions software version 3. Mobile phases
for LCMS analysis were HPLC grade or LCMS grade obtained from Sigma-Aldrich
and Fisher Scientific. The mobile phases consisted of a mixture LCMS
grade Acetonitrile/water (both with 0.1% formic acid for a pH of 2.7).
The initial setting for analysis was set at 5% acetonitrile (v/v),
then was linearly increased to 95% acetonitrile over 6 min. The gradient
was then held at 95% acetonitrile for 2 min and then linearly decreased
to 5% over 0.10 min and held until stop for an additional 1.90 min.
The total run time was equal to 12 min. The total flow rate was set
to 0.5 mL/min. The column oven and flow cell temperature for the diode
array detector was set at 30 °C. The auto sampler temperature
was held at 15 °C. 5uL was injected for analysis.