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Compound
Precursor
Ion
m/z
Quantifier
Ion
m/z
Qualifier Ion
m/z
Morphine
286.1
152.1
185.1
Morphine-
d
3
289.1
152.1
185.1
Oxymorphone
302.1
227.1
198.1
Oxymorphone-
d
3
305.1
230.1
201.1
Hydromorphone
286.1
185.0
157.1
Hydromorphone-
d
6
292.1
185.1
157.1
Codeine
300.2
152.1
165.1
Codeine-
d
3
303.1
152.1
215.1
Oxycodone
316.2
241.1
256.1
Oxycodone-
d
3
319.2
244.1
259.1
Hydrocodone
300.1
199.1
171.1
Hydrocodone-
d
3
303.1
199.1
171.1
Table 1. SRM transitions for opiates method
Compound
Precursor
Ion
m/z
Quantifier
Ion
m/z
Qualifier
Ion
m/z
7-Aminoclonazepam
286.1
222.1
250.1
7-Aminoclonazepam-
d
4
290.1
226.1
254.1
7-Aminonitrazepam
252.1
121.1
224.1
7-Aminoflunitrazepam
284.1
135.1
227.1
7-Aminoflunitrazepam-
d
7
291.2
138.2
230.1
α
-Hydroxytriazolam
359.0
331.0
239.0
Lorazepam
321.0
275.0
229.0
α
-Hydroxyalprazolam
325.0
297.1
229.0
α
-Hydroxyalprazolam-
d
5
330.1
302.1
216.1
Oxazepam
287.1
241.1
221.1
Oxazepam-
d
5
292.1
246.1
104.1
2-Hydroxyethylflurazepam 333.1
109.1
109.1
2-Hydroxyethylflurazepam-
d
4
337.1
215.1
113.1
Desalkyflurazepam
289.0
140.0
226.1
Desalkyflurazepam-
d
4
293.1
140.0
230.1
Temazepam
301.1
255.1
177.0
Temazepam-
d
5
306.1
260.1
177.0
Nordiazepam
271.1
140.1
208.1
Nordiazepam-
d
5
276.1
213.1
140.0
Alprazolam
309.1
281.1
205.1
Alprazolam-
d
5
314.1
286.1
210.1
Diazepam
285.1
193.1
154.0
Diazepam-
d
5
290.1
198.1
154.1
Midazolam
326.1
291.1
249.1
Midazolam-
d
4
330.1
295.1
253.1
Table 2. SRM transitions for benzodiazepines method
2
Experimental
Sample Preparation
Tables 1 and 2 contain the lists of opiates and
benzodiazepines analyzed. Sample preparation consisted
of glucuronide hydrolysis followed by dilution. For each
sample, a 200 µL aliquot of urine was spiked with 10 µL
of internal standards solution and 100 µL of
b
-glucuronidaze enzyme in an ammonium acetate buffer
(pH = 5.0). The samples were incubated at 60 °C for
2 hours. A 200 µL aliquot of methanol was added to each
sample to stop enzymatic reaction. Samples were cooled,
centrifuged, and diluted 20 times with deionized water.
Then, 20 µL of sample was injected into the liquid
chromatograph-mass spectrometer (LC-MS) system.
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
Prelude SPLC Multiplexing - Same Method on Both Channels
Injection 1
Injection 2
Channel 1
Injection 3
6 Opiates
(10 ng/mL each)
6 Opiates
0.0
1.0
2.0
3.0
4.0
5.0
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
Injection 1
Injection 2
Channel 2
Acquisition of relevent MS data
6 Opiates
6 Opiates
Figure 3. Analysis of six opiates (10 ng/mL) using both channels in multiplexed mode