TABLE III

Typical Operating Parameters, LC/UV/MS Analyses

Operating ParameterOperating value
ColumnAgilent Zorbax Eclipse Plus C18, 100 × 3.0 mm, 3.5 μm particles
Column Temperature40–50 °C
Mobile Phase ComponentsA = 10 mM ammonium acetate, B = acetonitrile
Mobile Phase GradientTime (min)% B
0.05.0
8.4100.0
35.0100.0
36.05.0
39.05.0
Mobile Stage Flow Rate0.8 mL/min
Injection Size10–50 μL
Detection, UV205–300 nm; spectra recorded at λ = 210, 220, 230, 250, and 270 nm; 205 nm for variable wavelength detector
Detection, MSAPI-ES or APCI, positive ion and negative ion (mass range 80–1500)
Instrumentation Used

  1. Agilent 1100 LC (vacuum degasser, binary pump, heated column compartment)/Model ATG1315A diode array detector/1100 mass selective detector (MSD). Agilent Chemstation rev A.10.02 software.

  2. Agilent 1100 LC (vacuum degasser, binary pump, heated column compartment, variable wavelength detector)/Applied Biosystems (Foster City, CA) Mariner TOF-MS (APCI mode). Agilent Chemstation rev A.10.02 and Applied Biosystems Data Explorer version 4.0.0.1 software.

  3. Agilent LCs (binary pump, autosampler, heated column compartment)/Thermo Finnigan (Waltham, MA) LCQ Deca XP Max ion trap MS or Thermo Finnigan LTQ FT Ultra MS or Waters Q-TOF Premier TOF-MS. Thermo Finnigan Xcalibur and Waters MassLynx software.

Sample PreparationNone, direct injection