Benzodiazepines in Urine on Raptor Biphenyl by LC-MS/MS

	Peaks	t_R (min)	Q1	Q3 Quantifier	Q3 Qualifier
1.	7-Aminoclonazepam	0.86	286.0	222.2	250.3
2.	7-Aminoflunitrazepam	1.15	284.2	135.2	240.2
3.	Flurazepam	1.29	388.1	315.1	288.2
4.	Midazolam	1.41	326.1	291.2	244.2
5.	N-Desmethylflunitrazepam	1.76	300.0	254.1	225.2
6.	Lorazepam	1.79	321.0	275.1	302.9
7.	Oxazepam	1.86	287.1	241.0	104.1
8.	Clonazepam	1.88	315.9	270.2	214.1
9.	Nitrazepam	1.88	282.1	236.1	207.2
10.	Desalkylflurazepam	1.93	289.0	140.1	165.1

	Peaks	t_R (min)	Q1	Q3 Quantifier	Q3 Qualifier
11.	2-Hydroxyethylflurazepam	2.00	333.1	246.1	166.0
12.	Nordiazepam	2.07	271.0	140.0	208.0
13.	Clobazam	2.13	301.1	259.1	224.0
14.	Estazolam	2.18	294.9	267.1	241.0
15.	Triazolam	2.19	343.1	308.2	315.0
16.	Flunitrazepam	2.19	314.4	239.1	211.0
17.	Temazepam	2.23	301.2	255.0	177.1
18.	Alprazolam	2.27	309.4	281.1	205.0
19.	Diazepam-d5 (IS)	2.45	290.0	154.1	-
20.	Diazepam	2.47	284.8	193.1	222.2

Column

Raptor Biphenyl (cat.# 9309A5E)

Dimensions:

50 mm x 3.0 mm ID

Particle Size:

2.7 µm

Guard Column:

Raptor Biphenyl EXP 5 mm, 3.0 mm ID, 2.7 µm (cat.# 9309A0253)

Temp.:

40 °C

Standard/Sample

Benzodiazepine custom mix

Conc.:

A 50 ng/mL standard was prepared in urine. 50 µL of 50 ng/mL urine standard was diluted with 950 µL of water prior to analysis (20x dilution).

Inj. Vol.:

20 µL

Mobile Phase

2 mM Ammonium formate and 0.2% formic acid in water

0.2% Formic acid in methanol

Time (min)	Flow (mL/min)	%A	%B
0.00	0.8	50	50
2.50	0.8	5	95
3.00	0.8	5	95
3.01	0.8	50	50
4.50	0.8	50	50

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