This application brief describes a solid phase extraction method for the analysis of benzodiazepines in serum using an Agilent AccuBOND SPE ODS (C18) 6 mL/500 mg cartridge.
The document provides a step-by-step procedure for preparing an SPE cartridge and processing serum samples to extract benzodiazepines. The process involves conditioning the cartridge with methanol and water, loading the sample mixed with sodium acetate, rinsing, and eluting with acetone before concentration and HPLC injection.
The guide specifies the required equipment, including AccuBOND ODS cartridges and vacuum manifolds, along with necessary reagents. It also includes chromatographic conditions for analysis, such as column specifications, mobile phase composition, flow rate, temperature, and detector settings.
| Cartridge Type | AccuBOND ODS (C18) 6 mL/ 500 mg |
|---|---|
| Column Type | C8, 15cm x 4.6 mm I.D. |
| Mobile Phase Composition | Acetonitrile/ Methanol/ 0.005M KH2PO4 (15/ 30/ 55) |
| Flow Rate | 2 mL/min |
| Column Temperature | 40°C |
| Detector Wavelength | 245 nm |
The document recommends preparing a 0.1 M solution of sodium acetate in water.
Vacuum should be left on for 30 seconds after all of the water has passed through the cartridge.
The residue should be concentrated using gentle heat below 45°C.