Contact an Account ManagerDirect Determination of Tramadol Glucuronides in Human Urine by HPLC with Fluorescence Detection
Summary
Type: SPE
Application #: 302600
Detection: HPLC
Conditions: Step 1:
Column conditioning: The cartridges were pretreated with 2,0 ml of methanol, 1,0 ml of water and 1,0 ml of water and 1,0 ml of 1,0 M ammonium sulphate adjusted to pH 9,8 with ammonia.
Column washing: with 1 ml of 1 M ammonium sulphate and 1 ml of water
Elution: with 2 ml of a mixture of 1,0 M phosphoric acid-methanol (7:3, v/v)
Step 2:
Column Conditioning: The cation-exchange cartridge was pretreated with 3 ml of methanol, 2 ml of water, and 1 ml of a mixture of 0,1 M phosphoric acid-methanol (7:3, v/v).
Sample application: The eluate (2 ml) from the C18 cartridge was passed through the cation-exchange cartridge followed by 1 ml of a mixture of 0,1 M phosphoric acid-methanol (7:3), 1 ml of 0,1 M acetic acid, and 2 ml of methanol.
Elution: The glucuronides were eluted with 2 ml of 3% ammonia in methanol.
Evaporation: The eluate was evaporated to dryness under a gentle stream of nitrogen. Fifty μl of the internal standard solution were added. After evaporation of the solvent the residue was dissolved in 1,0 ml of the mobile phase. Fifty μl were injected in the HPLC system.
Authors: Overbeck P Blaschke G
Source: Journal of Chromatography B 732 (1999) 185-192
Details
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