100 ng/mL Benzodiazepines in Hydrolyzed Urine on Ultra Biphenyl by LC-MS/MS

PeakstR (min)MRM1MRM2MRM3
1.7-Aminoclonazepam0.59286.0/250.3286.0/222.2286.0/121.0
2.7-Aminoflunitrazepam0.77284.2/135.2284.2/240.2284.2/226.1
3.Bromazepam1.10315.9/182.0315.9/209.1315.9/260.9
4.N-Desmethylflunitrazepam1.71300.0/254.1300.0/225.2300.0/198.0
5.Nitrazepam1.69282.1/236.1282.1/207.2282.1/180.0
6.Lorazepam1.71321.0/302.9321.0/275.1321.0/229.1
7.2-Hydroxyethylflurazepam1.79333.1/246.1333.1/166.0333.1/109.1
8.Nordiazepam1.78271.0/140.0271.0/208.0271.0/225.9
9.Estazolam1.88294.9/267.1294.9/241.0294.9/138.0
10.Desalkylflurazepam1.90289.0/140.0289.0/165.1289.0/226.0
11.Clonazepam1.92315.9/270.2315.9/214.1315.9/207.1
12.Alprazolam1.99309.4/281.1309.4/205.0309.4/274.1
13.Midazolam1.26326.1/291.2326.1/244.2326.1/209.0
14.Triazolam2.06343.1/315.0343.1/308.2343.1/239.2
15.Temazepam2.13301.2/283.1301.2/255.1301.2/228.1
16.Flurazepam1.34388.1/315.1388.1/288.2388.1/317.1
17.Flunitrazepam2.24314.4/211.0314.4/239.1314.4/268.1
18.Clobazam2.26301.1/259.1301.1/224.0301.1/181.1
19.Diazepam-D5 (IS)2.42290.0/154.1----
20.Diazepam2.45284.8/193.1284.8/222.2284.8/257.1
image
LC_CF0525
ColumnUltra Biphenyl (cat.# 9109552)
Dimensions:50 mm x 2.1 mm ID
Particle Size:5 µm
Pore Size:100 Å
Temp.:40 °C
Sample
Diluent:Starting mobile phase + 30 ng/mL IS
Conc.:100 ng/mL sample was diluted 20x prior to injection
Inj. Vol.:30 µL
Mobile Phase
A:Water + 2 mM ammonium formate + 0.2% formic acid
B:Acetonitrile + 10% water + 2 mM ammonium formate + 0.2% formic acid
Time (min)Flow (mL/min)%A%B
0.000.67030
3.00.64060
3.10.6595
3.50.6595
3.60.67030
4.5stop
DetectorAB SCIEX API 4000 MS/MS
Model #:API 4000
Ion Source:TurboIonSpray®
Ion Mode:ESI+
Ion Spray Voltage:2 kV
Curtain Gas:40 psi (275.8 kPa)
Gas 1:40 psi (275.8 kPa)
Gas 2:40 psi (275.8 kPa)
Source Temp.:600 °C
Mode:MRM
InstrumentApplied Biosystems/MDS Sciex LC-MS/MS System
NotesMS/MS instrument was operated in scheduled MRM mode.
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