AMP Line Intensity vs Time n10

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COC Line Intensity vs Time (n=10)

COC Line Intensity vs Time (n=10)

2 min -■—3 min -^—4 min -♦—5 min

Neg Cut-off 3x

Fig. 2. Line intensity studies. Neg = negative; 3x = three times the cutoff concentration of that particular drug. The intensity of each line is assigned an arbitrary number from 1 to 10, with the number 10 showing the darkest line and 1 representing the total absence of a line. AMP, amphetamine; COC, cocaine.

Table 2

Precision Study on the Oratect®

Drug concentration

Drug concentration

Table 2

Precision Study on the Oratect®

as % cut-off

0%

25%

50%

75%

300%

COC

20-

20-

20-

18-/2+

20+

OPI

20-

20-

20-

18-/2+

20+

THC

20-

20-

18-/2+

18-/2+

1-/19+

AMP

20-

20-

20-

18-/2+

20+

MET

20-

20-

20-

18-/2+

20+

PCP

20-

20-

20-

18-/2+

20+

BZO

20-

20-

20-

18-/2+

20+

- indicates negative result and + indicates positive result.

COC, cocaine; OPI, opiates; THC, tetrahydrocannabinol; AMP, amphetamines; MET, methamphetamines; PCP, phencyclidine; BZO, benzodiazepines.

- indicates negative result and + indicates positive result.

COC, cocaine; OPI, opiates; THC, tetrahydrocannabinol; AMP, amphetamines; MET, methamphetamines; PCP, phencyclidine; BZO, benzodiazepines.

results are summarized in Table 4. None of the subjects tested positive for PCP. Subjects who tested positive by Oratect for COC and OPI were likely to be tested positive by the urine drug screen. Oratect appeared to detect AMP better than urine test, and all subjects tested MET positive by Oratect were tested positive by Monitect. Confirmation of Oratect results by GC-MS ranged from 85 to 100% for COC, MET, AMP, and OPI.

All the volunteer subjects tested at the drug rehabilitation program appeared to have a job, and the tests were undertaken in the afternoon. It was suggested that this situation allowed little time for the test subject to smoke a marijuana cigarette immediately prior to being tested. Because the Oratect device tests for THC residue in the mouth cavity and the concentration of THC decreases rapidly after marijuana cigarette smoking (7), concentrations of THC might be too low to be detectable. The four samples that were detected positive by Oratect were all confirmed positive by GC-MS.

3.3. Correlation of Oratect Opiate Test With Oral-Fluid GC-MS and Urine Drug Screen

In another study (4), 10 normal human subjects were each administered one dose of Prometh with Codeine Cough Syrup (Alpharma USPD, Inc., Baltimore, MD) containing 10 mg of codeine phosphate, and their oral fluids and urine were analyzed for opiate after 1 h, 4 h, and 6 h. The urine samples were tested with Monitect PC11 Multiple Urine Drug Screen Test, while the oral-fluid samples were analyzed with three methods: Oratect screening test, Oratect confirmation test by GC-MS, and a laboratory-based test—Intercept oral-fluids

Table 3

Comparison of Enzyme-Linked Immunosorbent Assay (ELISA) and Oratect® Test Results

COC OPI PCP MET AMP

Table 3

Comparison of Enzyme-Linked Immunosorbent Assay (ELISA) and Oratect® Test Results

COC OPI PCP MET AMP

Drug level as multiple of C.O.

C.O. =

20 ng/mL

C.O. =

20 ng/mL

C.O. =

4 ng/mL

C.O. =

50 ng/mL

C.O. =

50 ng/mL

Aliquot #

ELISA

Oratect

ELISA

Oratect

ELISA

Oratect

ELISA

Oratect

ELISA

Oratect

1-11

6.OX to 1.5X

109-44

All +

79-35

All +

20-8

All +

171-75

All +

139-77

All +

12

1.3X

30

+»+

18

+»+

5

47

51

+»+

—A

13

1.2X

25

24

+»+

5

-

45

+»+

45

No

14

1.1X

40

+»+

17

4

+»+

58

43

+»+

15

1.0X

40

28

+»+

4

-

40

+»+

52

+»+

16

0.8X

57

14

4

36

+»+

30

17-26

0.7X to OX

23-0

All -

9-0

All -

3-0

All -

29-0

All -

24-0

All-

Oratect results are in duplicates. — indicates negative result, and + indicates positive result.

The ELISA results are the mean values of duplicate determinations and are expressed in ng/mL.

C.O., cut-off; COC, cocaine; OPI, opiates; PCP, phencyclidine; MET, methamphetamines; AMP, amphetamines.

Oratect results are in duplicates. — indicates negative result, and + indicates positive result.

The ELISA results are the mean values of duplicate determinations and are expressed in ng/mL.

C.O., cut-off; COC, cocaine; OPI, opiates; PCP, phencyclidine; MET, methamphetamines; AMP, amphetamines.

Table 4

Correlation Study of Urine Drug Screen and Oratect®

Table 4

Correlation Study of Urine Drug Screen and Oratect®

#

#

#

# Monitect®

Monitect

% Oratect

Drug

Monitect

Oratect

Both

positive/Oratect

negative/Oratect

GC/MS

detected

positive

positive

positive

negative

positive

confirmed

COC

36

34

32

4

2

90

MET

12

8

8

4

0

85

AMP

0

4

0

0

4

100

OPI

32

34

32

0

2

87

THC

28

4

4

24

0

100

PCP

0

0

0

0

0

N/A

GC-MS, gas chromatography-mass spectrometry; COC, cocaine; MET, methamphetamines; AMP, amphetamines; OPI, opiates; THC, tetrahydrocannabinol; PCP, phencyclidine.

GC-MS, gas chromatography-mass spectrometry; COC, cocaine; MET, methamphetamines; AMP, amphetamines; OPI, opiates; THC, tetrahydrocannabinol; PCP, phencyclidine.

confirmation test. The results as shown in Table 5 suggest that the Oratect device can detect the presence of codeine up to 6 h after the administration of a single 10-mg dose of codeine. It also confirms that GC-MS confirmatory testing using oral fluids collected with the Oratect device is a viable procedure and provides a correlation coefficient of 0.96 with the Intercept method.

4. Extension of Oratect Opiate Detection Window

When the Oratect opiate C.O. concentration was lowered to 10 ng/mL (6), the results (as shown in Table 6) suggested that the detection window can be extended twofold to 12 h. In this experiment, five healthy, normal subjects were each administered one dose of Prometh. Oratect test (with 10 ng/mL opiate C.O.) and Monitect urine drug screen were undertaken at 1,6, 8, 10, 14, and 16 h after the administration of the dose.

5. Detection of THC Over Time

Two men (subjects A and B) and one woman (subject C) were tested for THC after each of them smoked a single marijuana cigarette. Oratect drug screens and Intercept collections were performed at half-hour intervals. Both the Oratect collection pads and the Intecept devices were sent to Scientific Testing Laboratory for GC-MS analysis. The results as shown in Table 7 suggest that the Oratect can detect the presence of THC up to 2 h after use. Moreover, the Oractect GC-MS results correlate well with the Intercept GC-MS data.

6. Drug Interference Study

Drugs and metabolites that may potentially cross-react with the Oratect test were evaluated (8), and the results are shown in Tables 8 and 9. Each

Correlation of Oratect® Opiate Test With Oral-Fluid Gas Chromatography (GC)-Mass Spectrometry (MS) and Urine Drug Screen

1 h 4 h 6 h

Subj #

U

O

G

I

U

O

G

I

U

O

G

I

1

+

+

872

1084

+

+

30

28

+

+/-

18

18

2

+

+

192

200

+

+

20

34

+

+

ND

20

3

+

+

34

42

+

+

20

24

+

+/-

ND

ND

4

+

+

172

138

+

+

36

38

+

+

ND

ND

5

+

+

218

338

+

+

168

244

+

+

32

28

6

+

+

100

148

+

+

22

24

+

+

ND

18

7

+

+

104

154

+

+

24

42

+

+/-

16

20

8

+

+

64

78

+

+

28

26

+

+

16

22

9

+

+

84

106

+

+

38

60

+

+

30

28

10

+

+

196

554

+

+

50

84

+

-

12

38

U = Monitect® urine results; O = Oratect results; G = GC-MS confirmatory results using oral fluids collected with the Oratect device; I = GC-MS results using oral fluids collected with the Intercept device; ND = no drug detected.

U = Monitect® urine results; O = Oratect results; G = GC-MS confirmatory results using oral fluids collected with the Oratect device; I = GC-MS results using oral fluids collected with the Intercept device; ND = no drug detected.

Table 6

Oratect® Opiate Detection Window With Cut-Off at 10 ng/mL

Table 6

Oratect® Opiate Detection Window With Cut-Off at 10 ng/mL

#

U

O

U

O

U

O

U

O

U

O

U

O U O

1

+

+

+

+

+

+

+

+

+

+

+

— — -

2

+

+

+

+

+

+

+

+

+

+

+

+/- - -

3

+

+

+

+

+

+

+

+

+

+

+

+/- - -

4

+

+

+

+

+

+

+

+

+

+

+

- - -

5

+

+

+

+

+

+

+

+

+

+

+

+ - -

U = urine result; O = Oratect result.

U = urine result; O = Oratect result.

substance was dissolved in pooled normal negative saliva. Table 8 lists the drugs that do not interfere with Oratect results even at 100 ^g/mL concentration. Interfering compounds with concentrations that would cause a positive result are summarized in Table 9.

7. Potential Interference From Nondrug Sources

Several studies have evaluated the potential interference from nondrug sources.

Table 7

Detection of Tetrahydrocannabinol Over Time

Detection of Tetrahydrocannabinol Over Time

Table 7

Subject A

Subject B

Subject C

0 h

Oratect®

-

-

0.5 h

Oratect

+

+

+

Oratect GC-MS

494

512

393

Intercept™

253

322

211

1 h

Oratect

+

+

+

Oratect GC-MS

94.5

100

91

Intercept

NP

NP

NP

1.5 h

Oratect

+

+

+

Oratect GC-MS

48

65

50

Intercept

50

72

46

2 h

Oratect

+/-

+

+/-

Oratect GC-MS

34

44

33

Intercept

31

42

29

2.5 h

Oratect

-

+/-

-

Oratect GC-MS

NP

25

NP

Intercept

NP

28

NP

- indicates negative result; + indicates positive result; +/- indicates a borderline result; NP, test not performed; GC-MS, gas chromatography-mass spectrometry.

- indicates negative result; + indicates positive result; +/- indicates a borderline result; NP, test not performed; GC-MS, gas chromatography-mass spectrometry.

Table 8

Compounds That Do Not Cross-React at 100 |g/mL

Table 8

Compounds That Do Not Cross-React at 100 |g/mL

Acetaminophen

1-ephedrine

Acetylsalicyclic acid

Imipramine

Amobarbital

Lidocaine

Aspartame

Methadone

Buprenorphine

Pentobarbital

Caffeine

Phenobarbital

Chlorpromazine

d,1-phenylpropanolamine

Chloroquine

Propoxyphene

Desipramine

d,1-pseudoephedrine

Dextromethorphan

Secobarbital

7.1. Ethnic Meals

Thirteen volunteers from three ethnic groups including Caucasians, Asians, and Hispanics were tested with Oratect drug screens 2 h after the consumption of a meal typical of their ethnicities. This was repeated for three

Table 9

Interfering Compounds

ng/mL

Cocaine

Benzoylecgonine

20

Ecgonine Methylester

>5000

Anhydroecgonine

>5000

Methamphetamines

MDMA

200

Amphetamine

>5000

Ephredrine

>5000

Pseudoephredrine

>5000

MDA

>5000

Tetrahydrocannabinol

11-nor-A9-Tetrahydrocannabinol-9-carboxylic acid

20

A8-Tetrahydrocannabinol

1000

Amphetamine

MDA

100

Methmphetamine

>5,000

Ephredrine

>5,000

Pseudoephredrine

>5,000

MDMA

>5,000

Opiates

Codeine

50

6-Acetyl morphine

50

Ethylmorphine

50

Hydrocodone

100

Oxycodone

10,000

Phencyclidine

Phencyclidine

4

Benzodiazepines

Temazepam

10

Triazolam

15

Oxazepam

20

Diazepam

25

Benzodiazepines

Nitrazepam

30

Nordiazepam

40

Clobazam

100

Clonazepam

100

Flunitrazepam

150

Chlordiazepoxide

200

Prazepam

700

MDMA, methylenedioxymethamphetamine; MDA, methylenedioxyamphetamine.

MDMA, methylenedioxymethamphetamine; MDA, methylenedioxyamphetamine.

different days. Each meal consisted of dishes favored by the particular individual. A total of 36 test devices were used and all gave negative results, suggesting that food intake does not give false-positive results.

7.2. Cosmetic and Hygienic Products

Eight volunteers were tested with Oratect 30 min after brushing their teeth. They were again tested 30 min after applying lipstick and 30 min after smoking a cigarette. All 24 tests showed negative results, demonstrating that toothpaste, lipstick, and cigarettes do not interfere with the Oratect results.

7.3. Beverages

Oral fluids from drug-free volunteers were collected and pooled. Aliquots of the pooled oral fluids were each spiked with 10% v/v of the following beverages: (a) Lipton® tea; (b) black coffee; (c) Pepsi® Cola soft drink; (d) Dr. Pepper® soft drink; and (e) reconstituted Minute Maid® orange juice. When these spiked samples were applied to Oratect drug-screen devices, the test results indicated that none of the beverages studied gave false-positive results.

7.4. Food Ingredients

Aliquots of drug-free oral fluids were spiked with: (a) 10% v/v of 1 mg/mL solution of citric acid; (b) sugar with the final concentration of 1 mg/mL; (c) table salt with the final concentration of 1 mg/mL; and (d) monosodium glutamate with the final concentration of 1 mg/mL. When these spiked samples were applied to Oratect drug-screen devices, the results were all negative.

8. OratectPlus™: Expansion to Test for Alcohol

By incorporating into the Oratect device a dry reagent pad that accommodates the enzymatic reaction of alcohol oxidase and peroxidase, simultaneous determination of abused drugs and alcohol can be undertaken. This new configuration (Fig. 3), called the OratectPlus™ (9), uses the same procedure as the regular Oratect device except that the alcohol pad is read 2 min after the pink-purple flow appears in the test windows. The reported cut-off level was 0.04% blood-alcohol concentration, and the OratectPlus test results correlated well with an onsite alcohol test—AlcoScreen™ (Chematics, Inc. North Webster, IN).

9. Conclusions

The Oratect device has a simple design and an easy procedure for collection, testing, and confirmation sampling. Data presented here suggest that it

Fig. 3. OratectPlus® detecting abused drug and alcohol simultaneously.

can perform well and is a viable alternative to urine drug screen. To further the utility of the Oratect device platform, an alcohol pad has been incorporated so that drugs and alcohol can be detected simultaneously with the device. Such a development would further expand the use of Oratect.

References

1. Huestis MA and Cone EJ. Alternative testing matrices, in Drug Abuse Handbook (Karch SB, ed), CRC, Boca Raton, FL: 1998; 799-813.

2. Caplan Y and Goldberger B. Alternative specimens for workplace drug testing. J Anal Toxicol 2001;25:396-399.

3. Wong R. The current status of drug testing in the U.S. workforce. American Clinical Laboratory 2002;21, 21-23.

4. Mandatory Guidelines for Federal Workplace Drug Testing Programs, April 13, 2004 (69 FR 19644).

5. Wong B, Wong R, Fan P, and Tran M. Detection of abused drugs in oral fluid by an on-site one-step drug screen—Oratect™. Clin Chem 2003;49:A125.

6. Wong R, Wong B, and Tran C. Opiate time course study with Oratect™. Presented at the 41st International Meeting of the International Association of Forensic Toxi-cologists, Melbourne, November 2003.

7. Huestis MA and Cone EJ. Relationship of 89-tetrahydrocannabinol concentrations in oral fluid and plasma after controlled administration of smoked cannabis. J Anal Toxicol 2004;28:394-399.

8. Wong R, Wong B, Tran C, and Fan P. Detection of benzodiazepines by a one step onsite oral fluid rapid drug screen—Oratect™. Presented at the 41st International Meeting of the International Association of Forensic Toxicologists, Melbourne, 2003.

9. Wong R, Sook J, and Zolteck R. Alcohol testing by an onsite one-step oral fluid alcohol and drug combination test device—OratectPlus™. Presented at TIAFT/ SOFT Joint Congress, Washington, DC, September 2004.

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