Saliva Testing

The Science of Saliva Testing

Estrogens (estrone, estradiol and estriol), progesterone, testosterone, DHEA-S and cortisol are routinely measured in saliva at ZRT. Why saliva? Steroid hormones in the bloodstream are mostly (95-99%) bound to carrier proteins (hormone-binding globulins, albumin), and in this form they are unavailable to target tissues. Only the unbound fraction freely diffuses into tissues, including the salivary gland. Hormone levels in saliva therefore represent the quantity of the hormone that is currently available to target tissues and actively exerting specific effects on the body. Because of this, salivary hormone levels often relate to specific symptoms of hormone excesses or deficiencies. Research at ZRT has demonstrated clear correlations between salivary hormone levels and reported symptoms. The rationale for and clinical utility of saliva testing is well documented.

The very small concentrations of salivary hormones (only 1 – 5% of the total hormone levels that include protein-bound hormone found in serum) necessitate extremely sensitive assay methods. This is a particular issue for estrogens, which are present in very minute quantities in saliva, especially in older populations such as postmenopausal women. ZRT is unique as the only commercial laboratory using extracted saliva testing for estrogens. Extraction removes contaminants that interfere with the assay and concentrates the sample, significantly improving assay sensitivity compared to the “direct” assay methods available commercially14. In fact, poor correlations between serum tests and non-extraction salivary estradiol assays have unfortunately led to some skepticism about saliva testing. Also, because of the extremely sensitive assays, it is important to avoid blood contamination of saliva as a result of oral injury, therefore tooth brushing must be avoided before collecting saliva for testing. Saliva testing may also not be appropriate for sublingual hormone users unless samples are obtained at least 36 hours after the last dose. Blood spot testing is a preferred option for these patients.


  • Saliva testing, unlike serum tests, measures the bioavailable (“free”) levels of steroid hormones, correlating with symptomatology and potential deficiency
  • Samples are collected by the patient at home, allowing convenient timing of collection especially for cortisol, which must be measured at specific times of the day or night
  • Convenience of collection allows frequent sampling, e.g., during a menstrual cycle to determine fertility problems
  • Hormone levels can be assessed during topical hormone supplementation
  • Saliva collection avoids the stress of a blood draw, which can affect levels of cortisol
  • Hormones are stable in saliva at room temperature for up to 2 weeks, allowing for worldwide shipment and convenient mailing of samples for testing
  • Saliva testing is less expensive than conventional serum testing




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