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1,4-Benzodiazepines in Urine Analyzed by LC-MS using SPE.

Reference Number: AA-02318 Created: 05/01/2014 01:58 PM Last Updated: 09/18/2020 01:55 PM

Tamemazepam, Diazepam, Nordiazepam & Midazolam

Peak: 1. Temazepam 301.0739 m/z [M+H]+
         2. Diazepam 285.0790 [M+H]+
         3. Nordiazepam 271.0633 [M+H]+

         4. Midazolam 326.0855 [M+H]+

Method Conditions
Column: Cogent Diamond Hydride 2.0™, 2.2µm, 120Å
Catalog No.: 70200-05P-2 
Dimensions: 2.1 x 50 mm
Solvents: A: DI  H20  /  0 .1% formic  Acid (v/v)
B: Acetonitrile/ 0.1% formic Acid (v/v)
time (min.)     %B
   0       85
  6       70
  7       20
  9       20
        10      85

Injection vol.: 1 µL 
Flow rate: 0.4 ml/min 
ESI – POS  Perki Elme AxIO 2 TOF mass spectrometer
Extraction method: Spike urine  sampl was loaded into SPcartridge I (Clean Screen Xcel purchased from UCT BristolPA, USA) and eluted with 0.78 mL of acetonitrile, 200 microL of 2-propanol, 20 microL of ammonia. After the elution, thsampl wadried unde N2 gas and dissolved in 100 microof 50% methanol / 50% DI H2O / 0.1% formic acid. 

      Before injection, the 10 ppm spiked sample was filtered through a 0 .45µm nylon syringe filter (MicroSolv Tech Corp).

Discussion: The Cogent Diamond Hydride 2.o¯ column was successfully used in the analysis of 1,4-benzodiazepines in urine samples after SPE extraction. Four available compounds were well retained and separated. The procedure could be used for determination of this class of compounds in urine  samples  and other body fluids.



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