USES & APPLICATION OF
SALIVA FOR ROUTINE
DRUG TESTING
R. Sam Niedbala, Ph.D.
STC Technologies, Inc. Bethlehem, PA
BACKGROUND
SALIVA / ORAL FLUID AS A
TESTING MEDIUM
SALIVARY MOLECULES - Multifunctional
Saliva as a Diagnostic Fluid, Ann. N.Y. Acad. Sci. Vol. 694, 1993
WHAT IS ORAL FLUID?
 Whole Saliva: Combination of 3 glandular
fluids, some minor glands
Parotid: Serous Fluids
Sublingual: Mucous
Submandibular: Produces Both
 Mucosal Transudate: Fluid from Blood
Passive Transport: Through Crevicular Space
STC Technologies, Inc.
Capillary
Bed
Salivary
Gland
SALIVA FORMATION SCHEMATIC
Saliva as a Diagnostic Fluid, Ann. N.Y. Acad. Sci. Vol. 694, 1993
Potential Analytes For Saliva Testing
17-Hydroxyprogesterone
Acetaminophen
Amitriptyline
Amobarbital
Androstenedione
CA 19-9
Caffeine
Carbamazepine
Cholesterol
Cocaine
Corticosteroid Binding
Globulin
Cortisol
DHEA Binding Glycoprotein
Diazepam
Digoxin
Epidermal Growth Factor
Estradiol
Ethanol
Growth Hormone Release
Inhibiting Factor
HIV Antibodies
IgA
Insulin-Like Growth Factor I
Insulin-Like Growth Factor II
Lithium
Source: The Genesis Report/Dx,1991
Methaqualone
Nortryptyline
Phenobarbital
Phenytoin
Primidone
Progesterone
Secobarbital
Sex Hormone Binding
Globulin
Testosterone
Theophylline
Transforming Growth
Factor
Alpha
ORAL FLUID DOA PROCEDURE
COLLECTION
ARRIVE AT LAB
Test for IgG
IgG < 0.5 ng/L
IgG > 0.5 ng/L
Report Inadequate
Specimen
Screen EIA
Initial Positive
N egative
Confirmation
Report
N egative
Report Positive
Methods To Obtain Cleaner Saliva Samples
 Crevicular fluid -- filter paper strips between tooth
and gums
 Transudate fluid -- pad against gum and cheek
 Permeable membranes -- filter out high molecular
weight components of saliva
 Microaspiration -- very fine tipped aspirators for
crevicular fluid
 Salivary gland-induced -- “milking”
Source: The Genesis Report/Dx, 1991
Epitope, Inc. -- OraSure
ORAL FLUID COLLECTION
Saliva Flow Rates
Unstimulated: 0.32 mL/Minute Daily 1-1.5 L
Collector Dictates Final Volume
For Intercept
STEP 1
STEP 2
STEP 3
Saturated collector
Fluid is removed
Place Collector
is placed in fluid, sealed,
and analyzed
in Mouth
sent to lab
Dilution: 1:2 - 1:3 on average
Final units are based on ng/mL of saliva per device.
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SCREENING TECHNOLOGY
URINE VERSUS ORAL FLUID TARGETS
URINE
 Amphetamines
 Cocaine
 Opiates
 PCP
 THC





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ORAL FLUID
Amphetamines /
Methamphetamine
Cocaine / Benzoylecgonine
Codeine / Morphine /
MAM
PCP
THC / THC-COOH
HOW MICRO-PLATE EIA TESTS WORK
Add Substrate
Sample Reagent
Measure
Absorbance



YYY
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30 Minutes
Wash

30 Minutes
Stop
YYY
Y Antibody
 Conjugate
TEST ANALYTES + NIDA 5








Alcohol
Amphetamines / Methamphetamine
Barbiturates
Benzodiazepines
Cannabinoids (THC)
Cocaine
Opiates
Phencyclidine (PCP)
TYPICAL RESULTS
TECHNOLOGY:
IMMUNOASSAY CUTOFFS
DRUG CLASS
URINE
(ng/mL)
AMPHETS.
1000
COCAINE
MARIJUANA
OPIATES
PCP
300
50
300 (or 2000)
25
ORAL FLUID
(ng/mL)
SALIVA
(ng/mL)
AMPHET: 100 AMPHET: 400
METH: 40
METH 160
10
40
1
4
10
40
1
4
SALIVA CUT-OFFS CALCULATED BY MULTIPLYING ORAL FLUID BY A
FACTOR OF 4.
TECHNOLOGY:
CONFIRMATION CUTOFFS
DRUG
URINE
(ng/mL)
ORAL FLUID
(ng/mL)
SALIVA
(ng/mL)
AMPHET
METH
COCAINE
THC
OPIATES
PCP
500
500
150
15
300 (or 2000)
25
75
40
10
0.5
10
0.5
300
160
40
2
40
2
SALIVA CUT-OFF CALCULATED BY MULTIPLYING ORAL FLUID
CUT-OFF BY A FACTOR OF 4
ASSAY SUMMARIES -- VERSUS GC/MS
ORAL
FLUID
Cannabinoids Cocaine Methamphetamines Opiates
Sensitivity
100
97
100
100
Specificity
86
86
100
87.5
Urine
Cannabinoids
Cocaine Methamphetamines Opiates
Sensitivity
92
96
97.5
87
Specificity
94.4
100
92.7
100
COMPARISON TO URINE:
PERCENT POSITIVES
AMPS
COCAINE
THC
OPIATES
PCP
ORAL
FLUID
DOT
NONDOT
X: DOT
X: NONDOT
0.16
0.35
0.34
0.10
0.00
0.22
0.65
1.40
0.23
0.04
0.37
1.32
3.98
0.24
0.03
0.40
0.24
0.18
0.24
0.09
0.06
0.18
1.39
0.06
0.03
THC STUDY 1
TIME
URINE
ORAL FLUID
0
2
4
6
8
10
24
3/10
5/10
6/10
9/10
8/10
7/10
6/9
4/10
10/10
10/10
9/9
7/9
6/8
3/9
CUT-OFFS:
URINE 15 ng/mL
ORAL FLUID 1 ng/mL
THC STUDY 2
TIME
URINE
ORAL FLUID
0
0.5
1
2
4
24
7/10
7/8
9/9
10/10
10/10
10/10
0/10
10/10
9/10
9/10
7/10
2/10
CUT-OFFS:
URINE 15 ng/mL
ORAL FLUID 1 ng/mL
Window of Detection -- ORAL FLUID/URINE
SUBJECT #1 THC
ORAL FLUID (ng/mL)
20
110
Subject 1 - GC/MS/MS 100
90
Subject 1 - EIA +/80
70
60
50
40
30
20
Cutoff
GC/MS/MS 10
0
+
18
16
14
12
+
10
8
6
4
+
2
0
+
0
1
2
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URINE (ng/mL)
4
8
Hours
16
- - 24
48
72
+
Subject 1 - EIA
+/-
+
Cutoff EIA
- - -
- 0
1
2
4
8
Hours
16
24
48
72
Window of Detection -- ORAL FLUID/URINE
SUBJECT #2 THC
ORAL FLUID (ng/mL)
45
110
Subject 2 - GC/MS/MS 100
Subject 2 - EIA+/90
80
70
60
50
40
30
20
10
0
+
40
35
30
25
15
10
0
+ ++ ++ + + + +
Cutoff EIA
+
20
5
URINE (ng/mL)
+ +
Cutoff
-GC/MS/MS
0
1
2
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4
8
Hours
16
+ - +
24 48 72
Subject 2 - EIA
+/-
0
1
2
4
8
Hours
16
24
48
72
Window of Detection -- ORAL FLUID/URINE
SUBJECT #3 THC
ORAL FLUID (ng/mL)
50
45
110
Subject 3 - GC/MS/MS 100
Subject 3 - EIA+/90
80
70
60
50
40
30
20
10
0
+
40
35
30
25
20
15
10
5
0
+ +
Cutoff
- GC/MS/MS + - - - 0
1
2
4
8
16 24 48 72
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Hours
URINE (ng/mL)
Subject 3 - EIA
+/-
Cutoff EIA
-
0
1
- - -
2
4
8
Hours
16
24
48
72
COCAINE -- Typical Data
Screening Data (N = 500)
Positive Negative %Positive
Oral Fluid
50
450
10.0
Urine
62
438
12.4
70
Oral Fluid
+Oral Fluid/-Urine
+Urine/-Oral Fluid
Urine
60
50
40
30
20
10
0
Oral Fluid
+Oral Fluid/-Urine +Urine/-Oral Fluid
Urine
OPIATES -- Typical Data
Screening Data (N = 500)
Positive Negative %Positive
Oral Fluid
33
467
6.6
Urine
35
465
7.0
35
Oral Fluid
+Oral Fluid/-Urine
+Urine/-Oral Fluid
Urine
30
25
20
15
10
5
0
Oral Fluid
+Oral Fluid/-Urine
+Urine/-Oral Fluid
Urine
TIME OF DETECTION?
Literature Summary -Drugs of Abuse in Saliva/Urine
Drug Class
Cannabinoids
Cocaine
Phencyclidine
Opioids
Saliva Analyte

 THC

 THC

 THC

 THC

 THC

 THC

 THC
Cocaine
Cocaine
Phencyclidine
Morphine
Codeine
Hydromorphone
Methadone
Saliva Conc.
(ng/mL)
5-200
30-250
6-330
5-100
qual.
qual.
Saliva Time
of D etection
~ 7h
<4h
4-6 h
<8h
14 h
<6h
Urine Time of
D etection
5d
10 d
20 d
0.5
1-10
2-600
5-10 d
12-24 h
N /A
1-4 d
qual.
0.6
120
0.6
0.3-1
200
3-4 h
24 h
3h
36 h
4-10 h
24 h
8d
8-30 d
2d
Literature Summary -Drugs of Abuse in Saliva/Urine (Continued)
Drug Class
Saliva Analyte Saliva Conc.
(ng/mL)
Barbiturates
Amobarbital
200
Amobarbital
100
Hexobarbital
100
Phenobarbital
-8000
Methaqualone Methaqualone
20
300
Diazepines
Diazepam
2
Diazepam
700
Diazepam
20
Diazepam
2
Nordiazepam
692
N-Desmethyld
1100
Amphetamines Amphetamine
20-40
Saliva Time
of Detection
50 h
50 h
12 h
24 h
> 24 h
3h
50 h
<5h
24 h
8h
24 h
<5h
50 h
Urine Time
of Detection
1d
2 - 3 wk
3d
2d
INTERFERENTS
TYPICAL LAB -- URINE DATA
FROM 2.1.99 TO 4.30.99:
315,743 SPECIMENS TESTED
531 NITRITES (0.17%)
15 URINAIDS (0.01%)
129 PYRIDINE (0.02%)
196 SUBSTITUTED (0.06%)
TOTAL 871 SPECIMENS (0.28%)
AND 11,857 (3.8%) DILUTE SPECIMENS
Poppy Seed Effects -- Oral Fluid Vs. Urine
SUBJECT # 1
Time
Time
24 hrs.
8 hrs.
4 hrs.
24 hrs.
8 hrs.
4 hrs.
2 hrs.
1 hr.
0
0.25 hr.
0.5
2 hrs.
Signal
1
0 hr.
Signal
1.5
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
1 hr.
2
0.25 hr.
Urine
0 hr.
Oral Fluid
Poppy Seed Effects -- Oral Fluid Vs. Urine
SUBJECT # 2
Urine
2
Time
1
Time
24 hrs.
8 hrs.
4 hrs.
2 hrs.
1 hr.
0
0.25 hr.
0.5
0 hr.
24 hrs.
8 hrs.
4 hrs.
2 hrs.
1 hr.
Signal
1.5
0.25 hr.
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
0 hr.
Signal
Oral Fluid
Oral Fluid Results Following Poppy Seed Ingestion
Subjects consumed 40 grams of poppy seeds and then collected oral fluid and
urine specimens from 0.25 hours to 24 hours.
Subject
Time
(hours)
0.25
8-right
1
2
0.25
8-left
1
2
0.25
3-right
1
2
Reporting Limit
RSN:CK 9/8/99 DTABovr4.doc-193
Appendix
Codeine
Morphine
6-MAM
0
0
0
0
0
0
0
0
0
2 ng/mL
0
0
0
0
0
0
14
6
0
5 ng/mL
0
0
0
0
0
0
0
0
0
5 ng/mL
Oral Fluid Exposed to Poppy Seeds
Intercept™ Drugs of Abuse Collection Devices were spiked
with poppy seeds and stored at room temperature for 24 hours.
The oral fluid was then collected and analyzed by GC/MS.
Poppy Seeds
1
2
5
10
20
40 (20 mg)
40 mg
80 mg
Reporting
Limit
RSN:CK 9/8/99 DTABovr4.doc-193
Appendix
Codeine
0
0
0
0
13
0
15
22
2 ng/mL
Morphine
0
0
0
40
57
77
279
653
5 ng/mL
6-MAM
0
0
0
0
0
0
0
0
5 ng/mL
OPIATES
(ng/mL Oral Fluid)
Patient
Codeine Morphine 6-MAM Heroin
998
84
240
143
172
4
234
0
1005
2
1116
0
198
164
852
39
106
188
413
130
563
89
Reporting 2 ng/mL
Limit
RSN:CK 9/8/99 DTABovr4.doc-193
Appendix
333
246
77
14
98
14
472
123
345
419
233
5 ng/mL
119
0
166
0
11
0
5
0
28
0
0
0
304
0
91
0
149
0
382
33
248
0
5 ng/mL 5 ng/mL
Codeine/ 6-MAM/
Morphine Morphine
0.25
0.36
0.58
0.67
0.05
0.14
0.36
0.02
0.29
0.35
0.32
0.54
0.31
0.38
0.64
0.74
0.43
0.91
1.06
0
Time
24 hrs.
8 hrs.
4 hrs.
2 hrs.
1 hr.
0.25 hr.
0 hr.
0.5
0
Time
24 hrs.
1
8 hrs.
1.5
4 hrs.
2
2 hrs.
Oral Fluid
1 hr.
2
0.25 hr.
1.5
Signal
2.5
0 hr.
Signal
Hemp Oil Effects -- Oral Fluid Vs. Urine
Urine
1
0.5
CONCLUSIONS
New technology for screening &
confirmation make the use of oral
fluid possible.
Oral fluid provides overall similar
results as compared to urine.
STC Technologies, Inc.