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MS Conditions
Instrument:
LXQ ion trap mass spectrometer
Ionization:
ESI, Thermo Scientific Ion Max source
Capillary temperature:
275 ˚C
Spray voltage:
5.0 kV
Sheath gas:
30
Aux gas:
8
Data acquisition mode:
Polarity switching scan dependent experiment
Microscans:
1
WideBand Activation
™
:
On
Stepped Normalized
Collision Energy:
35% ± 10%
Method Validation and Results:
The method was prequalified by processing and analyzing
urine samples spiked with 10 randomly selected
compounds in concentrations of 10 ng/mL, 100 ng/mL
and 1000 ng/mL. Table 2 lists the concentration at which
each analyte in the toxicology screen for urine samples is
identified. The presence of an analyte at 10, 100 or 1000
ng/mL implies that the limit of detection is likely below
that value. Of the 275 compounds analyzed, 70% were
detected at 10 ng/mL, 20% at 100 ng/mL, 8% at 1000
ng/mL and 2% were detected at a concentration above
1000 ng/mL.
Chromatography
HPLC separation was performed with a Thermo Scientific
Accela pump using a Thermo Scientific Hypersil GOLD
PFP column (50 x 2.1 mm; 5 µm particles). Flow rate was
set to 200 µL/min. The gradient is summarized in Table 1
(solvent A = water/0.1% formic acid/10 mM ammonium
formate, solvent B = acteonitrile/0.1% formic acid).
Injection volume was 10 µL.
Table 1. Thirteen-minute LC method
Time (minutes)
%A
%B
0
95
5
0.5
95
5
5.5
5
95
8.5
5
95
8.6
5
95
13
95
5
11-Hydroxy-delta-9-THC
N
N
>1000
11-nor-9-carboxy-Delta-9-THC
N
N
P
2-Bromo-Alpha-Ergocryptine
P
P
P
2-Hydroxyethylflunitrazepam
N
P
P
3-Hydroxystanozolol
N
N
>1000
4-Hydroxynordiazepam
N
P
P
6-Acetylcodeine
P
P
P
6-Acetylmorphine (6-MAM)
P
P
P
7-Amino-Clonazepam
P
P
P
7-Amino-Flunitrozepam
P
P
P
Acebutolol
P
P
P
a-Hydroxy-Alprazolam
P
P
P
a-Hydroxy-Triazolam
P
P
P
Albuterol
P
P
P
alpha-Hydroxymidazolam
N
P
P
Alprazolam
P
P
P
Alprenolol
P
P
P
Aminorex
N
P
P
Amiodarone
P
P
P
Amitriptyline
P
P
P
Amlodipine
N
N
P
Amobarbital
P
P
P
Amoxapine
P
P
P
Amphetamine
P
P
P
Anhydroecgonine MethylEster
N
P
P
Antipyrine
N
N
>1000
Apomorphine
N
N
>1000
LXQ – 13 min method
Concentration Tested (ng/mL)
Compound
10
100
1000
All barbiturates require an APCI source for detection. P=Drug present.
N
=Drug not present.
Table 2. Results for spiked urine samples in toxicology screen by LC-MS/MS