1,4-Benzodiazepines in Urine Analyzed with ANP – AppNote

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ARTICLES, APPNOTES, FAQ (9292)
- AppNotes Library (1888)
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May 1, 2014
Bill Ciccone

AppNotes ANP/HILIC Methods / AppNotes Cogent Diamond Hydride / AppNotes Library / AppNotes Metabolites / ARTICLES, APPNOTES, FAQ

1,4-Benzodiazepine compounds from Urine Samples were analyzed 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 50mm
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.: 1µL
Flow rate: 0.4 ml / minute
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 0.78mL of acetonitrile, 200µL of 2-propanol, 20µL of ammonia.
—-After the elution, the sample was dried under N2 gas and dissolved in 100µL of 50:50 methanol / DI H2O / 0.1% formic acid.

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