Researchers are increasingly turning to saliva testing because it allows samples to be collected in a
convenient, non-invasive manner, and on a repeated basis. Anyone planning to use saliva testing must be
aware, however, that it is important to follow proper collection and handling procedures in order to insure
that the highest quality data is obtained. We offer the following advice based on our extensive experience
with saliva collection and testing.
Knowledge about saliva testing is rapidly growing and being revised, however, and there are still areas
where knowledge is limited, or where differences of opinion have not been resolved. We try to assist our
customers by providing them with up-to-date advice, but ultimately it is each customer’s responsibility to
make decisions about the best collection methods to use. We advise consulting the literature on the
analytes to be measured, and when the available literature is thin we strongly recommend a pilot study.
Preliminary Considerations
Variability of Salivary Analytes
Levels of many analytes in saliva do not remain static. Many steroid hormones are released into the
bloodstream in short bursts or pulses.(1) Because most steroids diffuse easily into saliva these variations
are also reflected relatively quickly in salivary concentrations. Salivary levels of some steroids, and other
analytes like α-amylase, also vary according to regular diurnal cycles.(2,3) Additionally, levels of some
salivary analytes are affected by stress or other stimuli.(4,5) It is therefore important to understand the
underlying physiology so as to be able to develop the
best strategies for sample collection. Several
factors may be of importance depending on the analyte of interest and the nature of the study:
• The diurnal cycle of the analyte must be understood. In most cases, sample collection should be made
at standardized times.
• The response and recovery characteristics of each analyte should be understood so that sample
collections are timed to properly capture responses.
• For analytes with pulsatile behavior we recommend collecting a minimum of three samples over a two
hour period. Equal volumes from each of the samples should be pooled to create one sample that
physically averages the fluctuations over that time period.
• Studies have shown that concentrations of SIgA and DHEA-S are affected by saliva flow rates.(6,7)
Ongoing research may eventually find other analytes that are also affected. For these analytes it is
necessary to record the saliva flow rate in order to express the assay results as a function of time.
3.Effect of Mouth Location on Salivary Analyte Content
Saliva is not a simple fluid with a consistent composition. Each of the major and minor salivary glands
has a different cellular makeup, and consequently there can be some differences in the composition of
saliva produced in each location. Levels of the steroid hormones measured by our kits are not affected by
the type of saliva collected, but a few analytes, such as SIgA and α-amylase, do show differences from
one gland to another.(8,9) For these analytes, researchers must be aware of the potential that absorbent
devices have to collect specific glandular saliva rather than whole, or mixed, saliva. Consistent placement
of the absorbent devices is important, and we offer recommendations below.
Research Participant Preparation and Documentation
In order to avoid the possibility of contaminating substances in the saliva that could interfere with the
immunoassay, we recommend the following precautions for research participants who will be donating
saliva:
• Avoid alcohol for 12 hours before sample collection.
• Do not eat a major meal within 60 minutes of sample collection.
• Avoid dairy products for 20 minutes before sample collection.
• Avoid foods with high sugar or acidity, or high caffeine content, immediately before sample collection,
since they may compromise the assay by lowering saliva pH and increasing bacterial growth.(12,13)
• Rinse mouth with water to remove food residue before sample collection, and swallow to increase
hydration.
• Wait at least 10 minutes after rinsing before collecting saliva to avoid sample dilution.
• Document consumption of alcohol, caffeine, nicotine, and prescription and over-the-counter
medications. (14-16)
• It is also advisable to document the physical activity level of research participants and the presence of
oral diseases. (17,18)
Sample Volume and Salivary Stimulants
Modern immunoassays are generally designed to work with small sample volumes, and in most cases
stimulants should not be required to collect adequate sample. We recommend against the use of oral
stimulants when collecting saliva samples, due to the possibility of causing assay interference or
alteration of levels of some analytes.(10) If stimulants are absolutely necessary they must be used
sparingly and in a consistent manner throughout the study.(11) Customers are encouraged to contact us
concerning the necessary sample volume based on the number and type of assays to be performed prior to
sample collection.
Blood Contamination in Saliva
Contamination of saliva samples with blood can also be a concern because the levels of most analytes are
higher in the general circulation than in saliva. Blood can leak into saliva under certain conditions,
4. including poor oral health, abrasive brushing, or injury, and even an invisible amount of blood
contamination has the potential to falsely elevate salivary analyte levels. (19-21) We recommend the
following:
• Research participants should not brush their teeth within 45 minutes prior to sample collection.
• Dental work should not be performed within 48 hours prior to sample collection.
• Research participants should be screened for oral health problems or injuries.
• Saliva samples visibly contaminated with blood should be discarded and recollected.
• Samples collected from populations that have little or no dental care, or known oral health problems,
may be screened with our Blood Contamination Assay Kit (Salimetrics Item No. 1-1302; 1-1302-5).
Saliva Pipetting Advice
Saliva contains mucus, which can make accurate pipetting difficult. Salimetrics advises freezing all
saliva samples once before performing the assay, followed by vortexing after the sample is thawed. This
procedure helps break up the mucus, and it can then be centrifuged into the bottom of the tube. Any other
cellular debris or food particles that were present are also removed in this step. Remove the sample for
testing from the clear solution, avoiding the pellet in the bottom of the tube. Due to the viscosity of
saliva, greater accuracy in sample volume is obtained by aspirating slowly, so as to avoid the formation of
bubbles.
Collection Methods and Devices:
Adults and Older Children
The selection of a collection method and device will depend on the analyte(s) of interest and the age of
the research participants. The following options are available:
Passive Drool
A very cost-effective method often used by our customers is the collection of whole saliva by passive
drool into a small vial. Passive drool is highly recommended because it is approved for use with almost
all analytes, unlike absorbent devices, which can sometimes cause interference in immunoassays. It is
important to use high-quality polypropylene vials, since other vials can lead to problems with analyte
retention or the introduction of contaminants that can interfere with the immunoassay. The vials used
must also seal tightly and be able to withstand temperatures as low as -80ºC. We sell 2 ml cryovials that
meet these requirements (Salimetrics Item No. 5002.01).
Materials required
Plastic drinking straws; Scissors; Cryovials (polypropylene, 2 mL capacity); Labels
1. Have research participants rinse their mouth with water 10 minutes prior to collection. Consult the
Research Participant Preparation and Documentation section above for additional advice.
2. Cut plastic drinking straws into 2-inch (5 cm) pieces.
3. Give each research participant one straw piece and one cryovial.
Instructions for Collecting Saliva
1. Instruct research participants to allow saliva to pool in the mouth. Some find it helpful to imagine
eating their favorite food.
2. With head tilted forward, research participants should drool down the straw and collect saliva in the
cryovial. (It is normal for saliva to foam, so we advise using a vial with twice the capacity of the desired
sample volume.)
3. Repeat as often as necessary until sufficient sample is collected. One mL (excluding foam) is adequate
for most tests. Collection of samples to be analyzed for more than one analyte may require larger vials.
Note: Secretory IgA and DHEA-S concentrations in saliva are affected by saliva flow rates. We
recommend recording the amount of time necessary to collect a given volume of saliva so as to
express the analyte measured as a function of time. Contact Salimetrics for details.
4. Keep samples cold after collection (4ºC) and freeze (-20º to -80ºC) as soon as possible.
Note: Freeze-thaw cycles should be minimized for some analytes. Contact Salimetrics for further
details.
The Salimetrics Oral Swab (SOS)
Some research participants are not willing or able to drool saliva into a vial. If the saliva samples are to
be analyzed for cortisol, testosterone, α-amylase, cotinine, C-reactive protein, or SIgA, the Salimetrics
Oral Swab (SOS) (Item No. 5001.02) is an excellent alternative to passive drool because of its ease of
use. The SOS is made of a non-toxic, inert polymer shaped into a 30 x 10 mm cylinder. It is not
recommended for children under the age of six, however, due to the possibility of a choking hazard.
When saliva is collected by placing the SOS underneath the tongue on the floor of the mouth, we find that
assay results are similar to those from whole saliva collected by passive drool. Under certain
conditions, however, there is a possibility that the SOS might collect specific glandular saliva, rather than
whole saliva. This could affect assay results for analytes such as α-amylase, CRP, and SIgA.
Researchers should be aware of this potential and decide on their collection strategy accordingly. The
SOS may also be used intentionally to collect samples from the parotid duct openings in the cheeks, as
directed below.
6
The SOS should be ordered with a Swab Storage Tube (Item No. 5001.01), which consists of a capped,
conical centrifuge tube with a separate insert that snaps into place inside the tube. The insert has a small
hole in its bottom, which allows the saliva to be centrifuged out of the swab into the bottom of the conical
tube. If centrifugation is not available, saliva from the swab may be expressed into a cryovial (Item No.
5002.01) using a needle-less 5 cc plastic syringe.
Instructions for Use
Note: We recommend placing the SOS into the tube insert shortly before distribution to the research
participants.
1. Remove SOS from tube leaving tube insert in place. Place into mouth as directed below. Keep in
place for 1-2 minutes. (If collecting from the parotid glands in the cheek saliva flow will be lower, and
collection time should be extended for up to 5 minutes to ensure adequate volume.)
Recommended SOS Placement
cortisol, cotinine,
testosterone
Under tongue
@-amylase (with
other analytes)
Under tongue
α-amylase* (alone) Between cheek and gum (near
upper 2nd molar)
SIgA†, CRP† Placement may vary depending on
focus of research
* Saliva from the parotid glands has higher concentrations of α-
amylase than pooled whole saliva from under the tongue
†Concentrations may vary depending on location in the mouth
2. Return SOS into tube insert.
3. Replace cap and snap securely onto tube.
7
4. Label the exterior of the tube using computer-generated, bar-coded labels provided by Salimetrics, or
waterproof pen. Position label so that the barcode lies horizontally along the length of the swab storagetube.
Note: Use labels recommended for freezing (cryolabels), not ordinary paper labels, which will fall off.
5. If samples cannot be frozen immediately, refrigerate or keep cool using insulated container with ice
packs. If swab storage box is used, place tubes in storage box cap side up.
6. We recommend freezing samples at or below -20°C within 1-2 hours of collection.
Note: Freeze-thaw cycles should be minimized for some analytes. Contact Salimetrics for further details.
7. On the day samples are to be assayed, bring them to room temperature and then centrifuge for 15
minutes at approximately 3,000 RPM (1500 x g). After centrifugation the tube insert and swab may
be discarded, but keep the cap. Assays should be performed using only clear saliva, avoiding any sediment that may have accumulated.
8. Re-centrifuge tubes following each freeze-thaw cycle since additional precipitates may develop upon refreezing.
9. Store unused swabs in the closed zip-lock bag under dry conditions (30-60% humidity) at room temperature.
Cautions:
• Do not use the SOS for children under the age of 6.
• Investigators using saliva samples collected with the SOS for biomarkers not approved by Salimetrics
do so at their own risk.
• Consult the section on Research Participant Preparation and Documentation above, and contact us
with any questions.
• The SOS may cause temporary dryness of mucosal membranes or oral cavity.
• Use only as directed.
Note: The SOS is made from an inert material that should theoretically pose no problem to specimens
stored frozen in the device. Studies of long-term storage at temperatures of -20 degrees C or colder
have shown no change in analyte stability over a period of two years. Nevertheless, before storage
for periods longer than two years we recommend that the specimen be removed from the SOS by
centrifugation or compression.
Salimetrics:
8. Collection Methods and Devices:
Infants and Small Children
Collecting saliva from infants and children under the age of six requires special consideration due to the
potential for choking when collection devices are placed in the mouth. A number of collection methods
listed below have been successfully employed, but it should be noted that these techniques are not
appropriate for use with all analytes.
Sorbettes
The Sorbette (Salimetrics Item No. 5029.00) is a small, arrowhead-shaped absorbent device attached to a
plastic shaft. We have approved it for collection of samples to be assayed for cortisol, α-amylase, cotinine,
and SIgA.
Note: SIgA levels are influenced by the saliva flow rate, which may be difficult to estimate when
collecting with this device.
The Sorbette is especially recommended when collecting samples from infants,(22,23) and it may also be of
use for older children under the age of six, or for bed-ridden elderly research participants. Sorbettes are used
in conjunction with conical tubes (Salimetrics Item No. 5001.04).
Instructions for Use
Supplies Needed: Sorbettes, conical tubes, sample ID labels.
1. Remove one Sorbette from envelope. Close envelope immediately to protect the remaining Sorbettes
from contact with moisture.
2. Put Sorbette under the tongue and allow it to absorb the whole saliva that pools there. Do not stop
collecting when the Sorbette begins to puff up. Leave it in place for at least 60 seconds, or preferably for 90
seconds to ensure the device is saturated.
Note: Due to its small size, the Sorbette has the potential to collect localized secretions from specific
areas of the mouth, which could affect results for analytes such as SIgA and alpha-amylase. We
therefore generally recommend that the Sorbette not be moved around in the mouth.
3. Under ideal circumstances, collection for 60-90 seconds yields approximately 200-300 uL of saliva. If
a greater volume is needed for testing, two Sorbettes may be held together by their shafts at the same
time. If taping the shafts together is necessary, be advised that the tape residue could potentially affect
assay results. We advise using a low-tack, removable tape. A better procedure would be to cut off the
contaminated portion of each shaft before placing the Sorbettes in the conical tube (see diagram below).
4. Sample collection from infants: The Sorbette can be placed under the tongue for 15-30 seconds at a time
and re-introduced as needed until the Sorbette is saturated (at least 60-90 seconds total time).
9
5. Place Sorbettes into the conical tube cap with the tips facing the cap end of the tube, as seen in the
following diagram. The Sorbette position is critical for recovering absorbed saliva during centrifugation.
SORBETTE SAMPLE PREPARATION
1) Place cap on a flat surface as shown.
2) Saturate one to three Sorbettes per research participant.
3) Insert Sorbettes tip down into cap.
4) Slide conical storage tube over purple sticks and snap down securely onto cap.
5) If conical tube storage box is used, place tubes in box with cap side up.
6. Label the exterior of the tube using computer-generated, bar-coded labels provided by Salimetrics,
or waterproof pen. Position label so that the barcode lies horizontally along the length of the swab
storage tube.
Note: Use labels recommended for freezing (cryolabels), not ordinary paper labels, which will
fall off.
7. If processing samples in-house, centrifuge the conical tube or cryovial for 15 minutes at 3000-3500 rpm
to extract the saliva. Remove the Sorbette with tweezers, then recap and proceed with testing.
Salimetrics
Study:
10
Notes:
• It is crucial that the saliva collected via Sorbette does not evaporate. Make sure the cap of the tube is snug
and sample is frozen within two hours after collection.
Note: Freeze-thaw cycles should be minimized for some analytes. Contact Salimetrics for further
details.
• It is best to centrifuge the saliva out of the Sorbette before freezing to lessen the chance of evaporation.
• Sorbettes are currently offered for research use only.
• Data from Sorbettes stored for 48 hours at -60°C, 4°C, and at room temperature show no statistically
significant differences in mean or CV (expressed as a proportion) from samples collected with a cotton
collection device.
• We discourage the Sorbette collection method for adults unless passive drool options prove impractical
(e.g., dementia patients).
Cotton rope
Cotton in various forms has frequently been used for saliva collection. Unfortunately, its use may be
problematic due to an unpleasant taste, the difficulty of recovering the sample and/or analyte from the
cotton, and the fact that it causes interference with certain biomarkers, including testosterone, SIgA,
estradiol, DHEA, and progesterone.(23,24) Still, for cortisol, α-amylase, or cotinine testing with infants,
3/8” (0.95 cm) diameter braided cotton rope (Salimetrics Item No. 5016.00) is an option.
Note: the cotton must be thoroughly saturated for accurate results.(23)
Instructions
1. Cut rope into 6” (15 cm ) lengths.
2. Hold one end of the rope firmly and place the other end into the infant’s mouth. Let the cotton rope
absorb saliva for approximately two or more minutes. Remove the rope and cut off the dry portion of the
rope. Place the wet portion into a needle-less 5cc syringe and squeeze the saliva into a cryovial.
3. Alternatively, you may place the wet cotton into the Saliva Storage Tube (Item No. 5001.01) normally
used for the SOS swab and centrifuge the saliva out of the cotton.
Note: Freeze-thaw cycles should be minimized for some analytes. Contact Salimetrics for further
details.
11
Collection Methods and Devices:
Non-Human Species
Analysis of hormones and other biomarkers in saliva is increasingly being used in an effort to monitor the
health and well-being of animals kept in confined conditions such as farms and zoos, or as household
pets. Salimetrics has provided EIA kits or testing services for studies that measure salivary cortisol in a
wide range of animals. These studies most often use cotton swabs or ropes to collect the saliva. Some
species, such as deer(25) and Guinea pigs,(26) have allowed plain cotton devices to be placed in their
mouths. Studies involving primates have often enticed the animals to lick or chew cotton pads or ropes
(attached to poles) by treating them with drink crystals, sugar solutions, or other flavorings.(27,28).
Highly trained police dogs have allowed their handlers to collect saliva with plain cotton swabs,(29) but
normal pet dogs have sometimes been encouraged to chew on cotton ropes by treating them with beef
flavor. One recent study using pet dogs examines the effects that such flavorings, or the use of citric acid
to stimulate saliva flow, may have on immunoassay results.(30)
A few papers have also experimented with other saliva collection devices for use with animals. The study
of pet dogs already cited reports that hydrocellulose microsponges (Sorbettes) appear to be useful,
offering a possible alternative to cotton. A study on CRP in pig saliva has reported that the swine
readily chew on larger sponges attached to poles. (31) Saliva has even been collected from large and
dangerous animals such as the rhinoceros by using a plastic spoon to scoop up several milliliters at a time
from the lower lip.(32)
The literature also contains numerous descriptions of techniques for saliva collection from mice and rats.
These include capillary tubes, filter paper strips, plastic pipettes, and more sophisticated suction devices.
These techniques are often used in conjunction with anesthesia, and they frequently use pilocarpine or
other chemicals to stimulate saliva flow. Salimetrics does not have direct experience with such methods
and cannot advise on their use.
Sample Handling and Storage
We encourage researchers to refrigerate or freeze samples as soon as possible after collection. Many
analytes are not stable at room temperature, and keeping samples cold after collection is important.
When samples remain at room temperature for periods of time longer than a few hours there is also
opportunity for bacterial growth, which can compromise assay validity.(33) We advocate a
conservative approach and advise that all samples should be maintained at 4ºC for no longer than
several hours before freezing them at or below -20ºC (temperature of a regular household freezer).
However, freeze-thaw cycles should be minimized for some analytes. It is critical that storage
conditions are researched prior to initiation of sample collection. Contact Salimetrics for details.
On the day samples are to be assayed, bring them to room temperature, vortex, and then centrifuge
for 15 minutes at approximately 3,000 RPM (1500 x g). Assays should be performed using only clear
saliva, avoiding any sediment present in the bottom of the tube. Re-centrifuge tubes following each
freeze-thaw cycle as additional precipitates may develop upon refreezing.
12
References
1. West, C. D., Mahajan, D. K., Chavre, V. J., & Nabors, C. J. (1973). Simultaneous measurement of multiple
plasma steroids by radioimmunoassay demonstrating episodic secretion. J Clin Endocrinol Metab, 36, 1230-36.
2. Dorn, L. D., Lucke, J. F., Loucks, T. L., Berga, S. L. (2007). Salivary cortisol reflects serum cortisol: analysis
of circadian profiles. Ann Clin Biochem, 44, 281-284.
3. Nater, U. M., Rohleder, N., Schlotz, W. et al. (2007). Determinants of the diurnal course of salivary alphaamylase.
Psychoneuroendocrinology, 32, 392-401.
4. Kreiger, D. T. (1975). Rhythms of ACTH and corticosteroid secretion in health and disease and their
experimental modification. J Steroid Biochem, 6, 758-791.
5. Rohleder, N., Nater, U. M. (In press, 2009). Determinants of salivary α-amylase in humans and
methodological considerations. Psychoneuroendocrinology.
6. Kugler, J., Hess, M., and Haake, D. (1992). Secretion of salivary immunoglobulin A in relation to age, saliva
flow, mood states, secretion of albumin, cortisol, and catecholamines in saliva. J Clin Immunol, 12(1), 45-9.
7. Vining, R. F., McGinley, R. A. and Symons, R. G. (1983). Hormones in saliva: Mode of entry and consequent
implications for clinical interpretation. Clin Chem, 29(10), 1752-56.
8. Crawford, J. M., Taubman, M. A., Smith, D. J. (1975). Minor salivary glands as a major source of secretory
immunoglobin A in the human oral cavity. Science 190 (4220), 1206-9.
9. Veerman, E. C., van den Keybus, P. A., Vissink, A. Nieuw Amerongen, A. V. Human glandular salivas: Their
separate collection and analysis (1996). Eur J Oral Sci 104(4), 346-52.
10. Granger, D. A., Kivlighan, K. T., Fortunato, C., Harmon, A. G., Hibel, L. C., Schwartz, E. B., & Whembolua,
G.-L. (2007). Integration of salivary biomarkers into developmental and behaviorally-oriented research:
Problems and solutions for collecting specimens. Physiology and Behavior, 92(4), 583-90.
11. Talge, N. M., Conzella, B., Kryzer, E. M., et al. (2005). It’s not that bad: error introduced by oral stimulants in
salivary cortisol research. Dev Psychobiol, 47(4), 369-76.
12. Klein, L. C., Whetzel, C. A., Bennett, J. M., Ritter, F. E., & Granger, D. A. (2006). Effects of caffeine and
stress on salivary alpha-amylase in young men: A salivary biomarker of sympathetic activity. Presented at the
annual meeting of the American Psychosomatic Society, Denver, CO.
13. Schwartz, E., Granger, D. A., Susman, E. J., Gunnar, M., & Laird, B. (1998). Assessing salivary cortisol in
studies of child development. Child Development, 69, 1503-1513.
14. Granger, D. A., Blair, C., Willoughby, M., Kivlighan, K. T., Hibel., L. C., Weigand, L. E., & Family Life
Project Investigators (2007). Individual differences in salivary cortisol and α-amylase in mothers and their
infants: Relation to tobacco smoke exposure. Developmental Psychobiology, 49(7), 692-701.
15. Hibel, L. C., Granger, D. A., Cicchetti, D., & Rogosch, F. (2007). Salivary biomarker levels and diurnal
variation: Associations with medications prescribed to control children’s problem behavior. Child
Development, 78, 927-937.
16. Hibel, L. C., Granger, D. A., Kivlighan, K. T., Blair, C., & The Family Life Project Investigators (2006).
Individual differences in salivary cortisol: Effects of common over the counter and prescription medications in
infants and their mothers. Hormones and Behavior, 50, 293-300.
17. Kivlighan, K. T. and Granger, D. A. (2006). Salivary α-amylase response to competition: Relation to gender,
previous experience, and attitudes. Psychoneuroendocrinology, 31(6), 703-14.
18. Henskens, Y. M. C., Van den Keijbus, P. A. M., Veerman, E. C. I., et al. (1996). Protein composition of whole
and parotid saliva in healthy and periodontitis subjects: Determination of cystatins, albumin, amylase and IgA.
J Periodont Res, 31, 57-65.
19. Schwartz, E., & Granger, D. A. (2004). Transferrin enzyme immunoassay for quantitative monitoring of blood
contamination in saliva. Clinical Chemistry, 50, 654-656.
13
20. Kivlighan, K. T., Granger, D. A., Schwartz, E. B., Nelson, V., & Curran, M. (2004). Quantifying blood leakage
into the oral mucosa and its effects on the measurement of cortisol, dehydroepiandrosterone, and testosterone in
saliva. Hormones and Behavior, 46, 39-46.
21. Granger, D. A., Cicchetti, D., Rogosch, F., Hibel, L. C., Teisl, M., & Flores, E. (2007). Blood contamination in
children’s saliva: Prevalence, stability, and impact on the measurement of salivary cortisol, testosterone, and
dehydroepiandrosterone. Psychoneuroendocrinology, 32(6), 724-33.
22. de Weerth, C., Jansen, J., Vos, M. H., Maitimu, I. & Lentjes, E. G. W. M. (2007). A new device for collecting
saliva for cortisol determination. Psychoneuroendocrinology, 32, 1144-1148.
23. Harmon, A., Hibel, L. C., Rumyantseva, O., & Granger, D. A. (2007). Measuring salivary cortisol in studies of
child development: Watch out–what goes in may not come out of commonly used saliva collection devices.
Developmental Psychobiology, 49, 495-500.
24. Shirtcliff, E. A., Granger, D. A., Schwartz, E., & Curran, M. J. (2001). Use of salivary biomarkers in
biobehavioral research: Cotton based sample collection methods can interfere with salivary immunoassay
results. Psychoneuroendocrinology, 26, 165-173.
25. Millspaugh, J. J., Washburn, B. E., Milanick, M. A., et al. (2002). Non invasive techniques for stress
assessment in white-tailed deer. Wildlife Society Bulletin, 30(3), 899-907.
26. Emack, J., Kostaki, A., Walker, C.-D., et al. (2008). Chronic maternal stress affects growth behavior and
hypothalamo-pituitary-adrenal function in juvenile offspring. Hormones and Behavior, 54, 514-20.
27. Lutz, C. K., Tiefenbacher, S., Jorgensen, M. J., et al. (2000). Techniques for collecting saliva from awake,
unrestrained adult monkeys for cortisol assay. Am J Primatol, 52, 93-99.
28. Newman, J. L., Perry, J. L., Carroll, M. E. (2007). Social stimuli enhance phencyclidine (PCP) selfadministration
in rhesus monkeys. Pharmacology, Biochemistry and Behavior, 87(2), 280-88.
29. Horvath, Z., Igyártó, B.-Z., Magyar, A. & Miklósi, Á. (2007). Three different coping styles in police dogs
exposed to a short-term challenge. Hormones and Behavior, 52, 621-30.
30. Dreschel, N. A. & Granger, D. A. (2009). Methods of collection for salivary cortisol measurement in dogs.
Hormones and Behavior, 55, 163-8.
31. Gutiérrez, A. M., Martínez-Subiela, S., Eckersall, P. D., Cerón, J. J. (in press, 2008). C-reactive protein
quantification in porcine saliva: A minimally invasive test for pig health monitoring. Veterinary Journal.
32. Gómez, A., Jewell, E., Walker, S. & Brown, J. (2004). Use of salivary steroid analyses to assess ovarian cycles
in an Indian rhinoceros at the National Zoological Park. Zoo Biology, 23, 510-12.
33. Whembolua, G.-L., Granger, D. A., Kivlighan, K. T., Marguin, J. A. & Singer, S. (2006). Bacteria in the oral
mucosa and its effects on the measurement of cortisol, dehydroepiandrosterone, and testosterone in saliva.
Hormones and Behavior, 49, 478-483.
14
Approved Saliva Collection Methods
by Analyte
Adults &
Children 6 or older
Infants &
Children under 6
Analyte Passive
Drool SOS Cotton Sorbette Cotton
Rope
Alpha-amylase
Androstenedione
CgA
Cortisol
Cotinine
C-Reactive Protein
Dexamethasone
DHEA
DHEA-S
Estradiol
Estriol
Estrone
IL-6
Melatonin
Neopterin
17-OH-Progesterone
Progesterone
Secretory IgA
Testosterone
Total Protein
Transferrin
(Blood Contamination)
(Shaded cells indicate approved methods)Note: concentrations are affected by saliva flow rate Assay validated only with cotton collection
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