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ANALYTICAL EVALUATION OF FIVE ORAL FLUID
DRUG TESTING DEVICES
Cristina Isalberti, Sylvie Van Stechelman, Sara-Ann Legrand, Trudy Van der Linden, Alain Verstraete
Department of Clinical Chemistry, Microbiology and Immunology, Ghent University, De Pintelaan 185, 9000 Ghent, Belgium
INTRODUCTION
TESTED DEVICES
Varian OraLab ® 6 - Consists of a collection stick and an expresser
vial. Testing starts with oral fluid collection and results can be read in
10 - 15 minutes.
The correlation between drug presence in blood and oral fluid and the ease of sample collection make oral
fluid an ideal matrix for roadside drug tests aimed at detecting “driving under the influence” [1, 2].
The possibility of using the result of oral fluid tests to detect impaired drivers was introduced in the Belgian
Traffic Legislation in July 2009 [3].
A reliable rapid on-site oral fluid testing device, that can be used routinely in police controls, is the basis for
implementing the law.
Dräger DrugTest ® 5000 - Comprises the Dräger
DrugTest 5000 Analyzer and a test kit. The test kit consists
of a test cassette with an oral fluid collector. Once the test
cassette and the cartridge are placed into the analyzer
results are shown within a few minutes.
Cozart ® DDS 806 - Comprises a collector swab, a buffer bottle, a
disposable test cartridge and a handheld instrument for result
interpretation. Two drugs can be tested in 90 seconds, and 5/6 drug
classes in 5 minutes.
AIM
The purpose of this study was to evaluate the performance of five oral fluid testing devices (Varian OraLab ®
6, Dräger DrugTest® 5000, Cozart® DDS 806, Mavand RapidSTAT®, Innovacon OrAlert) that may be used in
roadside drug tests and assess their suitability for detecting cocaine, opiates, cannabinoids and amphetamine
at the cut-offs set by the Belgian legislation.
Mavand RapidSTAT ® - Consists of a collector stick, a buffer
bottle and the test panel. The test results can be read either
straight from the test device or by inserting the device into a
mobile reader. The total time needed for testing is 7-12
minutes.
Short descriptions of the tested devices are reported in the box to the right. Detailed descriptions, including
classes of substances detected, cut-offs and operating procedures, can be found in Deliverable 3.2.2 on the
DRUID project official website [4].
Innovacon OrAlert - Consists of a collector stick and of a test
device. An oral fluid sample is collected during the testing and can be
sent to laboratory for confirmation. The total test time is approximately
10 minutes.
METHODS
More than 760 oral fluid samples were collected from volunteers either at drug addiction treatment centres or during roadside sessions.
At the time of collection volunteers provided two oral fluid samples. One was tested on-site with one of the selected devices, while the second sample was used for confirmation
analysis by ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS) after liquid-liquid extraction (conditions described below).
Sensitivity, specificity, accuracy and prevalence for cocaine, opiates, cannabinoids and amphetamine were calculated for each device applying Belgian legal confirmation cut-offs (THC
10 mg/mL; amphetamine 25 ng/mL; MDMA 25 ng/mL; free morphine or 6-AM 5 ng/mL; cocaine or benzoylecgonine 10 ng/mL).
UPLC-MS/MS CONFIRMATION ANALYSIS
The oral fluid samples for confirmation analysis were collected using StatSureTM Saliva Sampler TM device. This consists of an absorptive pad with a volume indicator that turns blue
when 1 mL of oral fluid has been collected, and a plastic tube containing 1 mL of buffer solution. To account for any possible variability in the volume of oral fluid collected, drug
concentrations obtained after UPLC-MS/MS analysis were corrected on the basis of the average weight of an empty StatSureTM device and the weight of the StatSureTM device after
sample collection.
For the liquid-liquid extraction 20 mL of 200 ng/mL isotope-labelled internal standards solution and 200 mL of NH4HCO3 (0.2 M, pH 9.3) were added to 400 mL of sample from the
StatSureTM Saliva Sampler. Samples were then extracted with 1.25 mL of heptane/ethyl acetate (1:4). After 15 minutes on a shaker and centrifugation, the organic phase was
transferred into a clean test tube and evaporated to dryness at room temperature. Samples were reconstituted in 100 mL methanol/water (50:50) and transferred into a vial for UPLCMS/MS analysis. Calibration curves (range 0.5-200 ng/mL) and quality controls, made by spiking target compounds in drug free oral fluid samples collected using StatSureTM Saliva
Sampler TM device, were extracted alongside each batch of samples.
UPLC-MS/MS analysis were performed on an AcquityTM ultra performance liquid chromatograph (Waters) equipped with an Acquity UPLC BEH C18 column (1.7µm, 2.1 x 100mm) and
a Vanguard BEH C18 pre-column (1.7µm, 2.1 x 5mm). Detection was performed using a Waters Quattro Premier XE tandem mass spectrometer operating in MRM mode. UPLC and
MS conditions are reported in the tables below.
UPLC gradient elution
Target
Q1
6-acetylmorphine
6-acetylmorphine-D3
Amphetamine
Amphetamine-D5
Benzoylecgonine
Benzoylecgonine-D3
Cocaine
Cocaine-D3
Codeine
Codeine-D3
MDA
328.1
331.1
136.1
141.0
290.1
293.1
304.1
307.1
300.1
303.1
180.0
Collision
RT Cone
Q2
energy
(min) (V)
(eV)
152.1 3.61
47
61
164.9 3.58
45
37
119.1 3.43
15
9
92.9 3.37
17
27
168.0 2.64
33
19
171.0 2.65
33
19
182.1 4.66
31
19
185.0 4.64
31
19
165.0 3.85
41
43
215.0 3.83
45
25
105.0 3.28
15
21
Target
Q1
MDA-D5
MDEA
MDEA-D5
MDMA
MDMA-D5
Metamphetamine
Methamphetamine-D5
Morphine
Morphine-D3
THC
THC-D3
185.0
208.1
213.1
194.1
199.1
150.0
155.0
286.1
289.1
315.2
318.1
Collision
RT Cone
Q2
energy
(min) (V)
(eV)
110.0 3.23
17
21
163.0 3.79
23
13
162.9 3.69
21
13
163.0 3.43
21
13
135.2 3.34
21
21
91.0 3.61
21
17
120.9 3.55
19
11
152.1 3.13
45
53
164.9 3.10
43
37
193.1 5.44
31
21
196.0 5.43
31
25
General parameters for mass spectrometry
ES + Source Capillary Voltage
0.8 kV
Extractor Voltage
4V
RF Lens
0.0 V
Source Temperature
140°C
Desolvation Temperature 450°C
Desolvation Gas Flow
1,000 L/h
Cone Gas Flow
50 L/h
Analyser
Collision Gas Flow
0,15 mL/min
Time
Flow
%A
%B
(min)
(mL/min)
0.00
0.40
90.0
10.0
1.00
0.40
90.0
10.0
1.50
0.40
60.0
40.0
2.99
0.40
33.8
66.2
3.00
0.25
33.8
66.2
3.80
0.25
25.0
75.0
4.30
0.40
5.0
95.0
5.30
0.40
5.0
95.0
5.50
0.40
90.0
10.0
7.00
0.40
90.0
10.0
A = NH4HCO3 (2mM, pH 9.3); B = MeOH
Column temperature = 60°C
Volume injected = 25 mL
RESULTS
CONCLUSIONS
Specificity, sensitivity, accuracy and drug prevalence for each device are shown in the table below.
In July 2009 the possibility of using oral fluid drug testing devices to
target impaired drivers was introduced in the Belgian legislation.
DEVICE
Varian
OraLab ®
6
Dräger
DrugTest ®
5000
Cozart ®
DDS 806
Mavand
RapidSTAT
®
Innovacon
OrAlert
TARGET
substance
Cocaine
Opiates
THC
Amph
Cocaine
Opiates
THC
Amph
Cocaine
Opiates
THC
Amph
Cocaine
Opiates
THC
Amph
Cocaine
Opiates
THC
Amph
True
pos
19
85
18
19
6
75
20
6
1
49
11
4
3
62
13
1
7
64
3
1
False
pos
0
2
2
0
1
3
5
0
1
0
0
1
3
2
12
4
0
2
0
10
True
neg
195
120
159
216
124
45
107
129
129
66
99
131
120
52
91
123
96
20
97
97
False
neg
35
42
70
14
6
14
5
2
7
23
28
2
7
17
17
5
7
24
10
2
N. of
tests
249
249
249
249
137
137
137
137
138
138
138
138
133
133
133
133
110
110
110
110
Sens.
(%)
35.2
66.9
20.5
57.6
50.0
84.3
80.0
75.0
12.5
68.1
28.2
66.7
30.0
78.5
43.3
16.7
50.0
72.7
23.1
-
Spec.
(%)
100
98.4
98.8
100
99.2
93.8
95.5
100
99.2
100
100
99.2
97.6
96.3
88.3
96.9
100
90.9
100
90.7
Acc.
(%)
85.9
82.3
71.1
94.4
94.9
87.6
92.7
98.5
94.2
83.3
79.7
97.8
92.5
85.7
78.2
93.2
93.6
76.4
90.9
89.1
Prev.
(%)
21.7
51.0
35.3
13.3
8.8
65.0
18.2
5.8
5.8
52.2
28.3
4.3
7.5
59.4
22.6
4.5
12.7
80.0
11.8
2.7
The aim of this study was to evaluate the reliability of 5 oral fluid
testing devices to determine whether any of them may be suitable to
detect drug-impaired drivers during roadside police controls. The
evaluation was carried out taking the Belgian law confirmation cut-offs
for cocaine, opiates, cannabinoids and amphetamine in consideration.
All tested devices showed good specificity for all drug classes.
Sensitivity and accuracy were very different among devices and drug
classes. In particular, when applying Belgian low cut-offs, all devices
showed unsatisfactory sensitivity for cocaine; the highest being equal
to 50% for both Dräger DrugTest ® 5000 and Mavand RapidSTAT
devices.
Considering that cannabis, followed by amphetamines and ecstasy, is
the most prevalent drug among impaired drivers in Belgium [5], only
Dräger DrugTest ® 5000 appeared to be sensitive enough to be used
during roadside police controls.
REFERENCES
[1] Aps JK, Martens LV (2005) Forensic Sci Int 150(2-3): 119-131 - [2] Verstraete AG (2005) Forensic Sci Int 150: 143-150 - [3]
Belgisch Staatsblad/Moniteur Belge (15.09.2009 – Ed. 2): 62185-62189 - [4] www.druid-project.eu - [5] Raes E, Verstraete AG
(2005) J Anal Toxicology 29: 632-636
DISCLAIMER - This abstract has been produced under the project “Driving Under the Influence of Drugs, Alcohol and Medicines” (DRUID) financed by the European Community within the framework of the EU 6th Framework Program. This abstract reflects only
the authors’ view. The European Community is not liable for any use that may be made of the information contained therein.