This application note describes the use of the Agilent 1100 Series LC system for the analysis of β-blockers in blood and other biological fluids using online and offline sample preparation methods.
The document details a method for analyzing beta-blocking drugs (pindolol, timolol, metoprolol, and propranolol) in biological fluids such as urine, plasma, and tissue. It addresses the challenges of sample preparation, which is often labor-intensive, by evaluating various techniques including protein precipitation and liquid/liquid extraction.
| System Used | Agilent 1100 Series LC system |
|---|---|
| Detectors Used | UV, fluorescence and mass spectrometry detection |
| Pump Type (Standard) | Agilent 1100 Series binary pump |
| Autosampler Type (Standard) | Agilent 1100 Series well-plate autosampler |
| Column Thermostat Type (Standard) | Agilent 1100 Series column thermostat |
| Detector Type (Standard) | Agilent 1100 Series diode-array detector |
| Mass Spectrometer (Standard) | Agilent LC/MSD quadrupole mass spectrometer with ESI source |
| Detector Type (Capillary) | Agilent 1100 Series diode-array detector equipped with a 500-nL cell |
| Mass Spectrometer (Capillary) | Agilent XCT Ion Trap mass spectrometer with ESI source |
| Column (Cleaning) | 30 x 4.6 mm ZORBAX XDB C-18, 5-pm |
| Column (Gel Filtration) | 100 x 2.1 mm Poly LC INC Polyhydroxyethyl A, 3 pm, 500A |
| Column (MARS) | 4.6 x 50 mm, Hv-6 |
| Column (Analytical Capillary) | 100 x 0.8:mm ZORBAX SB C-8, 3.5-4pm |
| Column (Analytical Narrow-Bore) | 75 x 0.32mm ZORBAX SB C-8, 3.5-4pm |
| Column (Analytical Capillary) | 50 x 0.3:mm ZORBAX SB C-8, 3.5-4pm |
| Column (Analytical Narrow-Bore) | 30 x 2.l4mm ZORBAX SB C-8 |
| Column (Analytical Capillary) | 50 x 0.3 mm ZORBAX SB C-8 |
| Mobile Phase A (Analytical) | water+ 0.05 % FA, acetonirile + 0.045 % FA |
| Flow Rate (Analytical) | 0.01 mL/min |
| Injection Volume (Analytical) | 1 pL with needle wash in flush port |
| Source Type (MSD) | ESI |
| Gas Temperature (MSD) | 350 °C |
| Drying Gas (MSD) | 5 Limin |
| Nebulizer Pressure (MSD) | 15 psig |
| Veap Voltage (MSD) | 4000 V positive |
The LOD is between 200 and 400 pg for a spiked urine sample when using the MSD.
Recovery rates for urine are between 70 and 90 %.