1,4-Benzodiazepines from Urine Samples were analyzed using a Cogent Diamond Hydride 2.o HPLC Column successfully after Solid Phase Extraction (SPE).

Four available Compounds (shown below) were well retained and separated. The procedure could be used for determination of this class of compounds in urine samples and other body fluids.

Peaks:
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)
Gradient:   

    time    %B
     0      85
     6      70
     7      20
     9      20
    10      85

Injection vol.: 1 µL
Flow rate: 0.4 ml/min
Detection: ESI – POS – Perkin Elmer AxION 2 TOF Mass Spectrometer
Samples: Extraction method: Spiked urine sample was loaded into SPE cartridge I (Clean Screen Xcel™ purchased from UCT Bristol, PA, USA) and eluted with 0.78 mL of acetonitrile, 200 microL of 2-propanol, 20 microL of ammonia. After the elution, the sample was dried under N2 gas and dissolved in 100 microL of 50% methanol / 50% DI H2O / 0.1% formic acid.

Before injection, the 10 ppm spiked sample was filtered through a 0 .45µm AQ Brand Nylon Syringe Filter (MicroSolv Tech Corp).


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