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Salivary Hormone Testing

Saliva testing is an easy and noninvasive way of assessing your patients hormone balancing needs, and is proving to be the most reliable medium for measuring hormone levels. Unlike serum tests, saliva testing represents only hormones actively delivered to receptors in the body. Clinically, it is far more relevant to test these bioavailable hormones and provide an accurate reflection of the body’s active hormone levels.

Saliva Test Tubes

Appreciating the reliability of saliva testing is based on understanding the difference between steroid hormones in saliva and in serum. This difference is based on whether or not the hormones are bound to proteins in the medium used for testing. The majority of hormones exist in one of two forms: free (5%) or protein bound (95%). Only the free hormones are biologically active, or bio-available, and available for delivery to receptors in the body. Protein bound hormones do not fit the receptors and are considered non-bioavailable. When blood is filtered through the salivary glands, the bound hormone components are too large to pass through the cell membranes. Only the unbound hormones pass through and into the saliva. Saliva testing measures the bioavailable hormone - the clinically relevant portion delivered to the receptors in the tissues of the body.

Salivary hormone levels are expected to be much lower than serum levels, as only the unbound hormones are being measured. When healthcare providers measure serum hormone levels and prescribe hormone replacement therapy based on those results, patients are often overdosed. If the patients are then tested using saliva, the results are extraordinarily high and create confusion resulting from a lack of correlation between the two methods.

This discrepancy becomes especially important when monitoring topical, or transdermal, hormone therapy. Studies show that this method of delivery results in increased tissue hormone levels (thus measurable in saliva), but no parallel increase in serum levels. Therefore, serum testing cannot be used to monitor topical hormone therapy.

Saliva measures the "unbound" biologically active or free hormone levels in the body:

When blood is filtered through the salivary glands, the bound hormone components are too large to pass through the cell membranes of the salivary glands. Only the unbound hormones pass through and into the saliva. What is measured in the saliva is considered the "free", or bioavailable hormone, that which will be delivered to the receptors in the tissues of the body.

Serum measures the "protein bound" biologically inactive hormone levels in the body:

In order for steroid hormones to be detected in serum, they must be bound to circulating proteins. In this bound state, they are unable to fit into receptors in the body, and therefore will not be delivered to tissues. They are considered inactive, or non-bioavailable.

Only saliva testing measures topically dosed hormones:

The discrepancy between free and protein bound hormones becomes especially important when monitoring topical, or transdermal, hormone therapy. Studies show that this method of delivery results in increased tissue hormone levels (thus measurable in saliva), but no parallel increase in serum levels. Therefore, serum testing cannot be used to monitor topical hormone therapy.

Hormones are powerful molecules essential for maintaining physical and mental health. We frequently think of estrogen as being a female hormone, and testosterone as being a male hormone. But men AND women make both, plus several more that need to be in balance for optimum health. An imbalance of any one hormone can throw your physical and mental health out of balance, causing aggravating and even serious health problems.

One size does not fit all when it comes to hormones! For decades western medicine has prescribed hormone replacement therapy as if everyone needed the same thing and the same amount. Nothing could be further from the truth. Your hormones are like your fingerprints and in order to achieve optimal health, you need to know what your specific imbalances are.

There are several ways to test for hormones (saliva, serum and urine), but the state-of-the-art method is through saliva. This is because only the active portions of hormones are measured and it is these portions that determine how individuals feel. So if your patient is seeking bio-identical hormone replacement (BHRT), youll need to know active hormone levels. In addition, if using a topical (transdermal) hormone preparation for treatment, saliva testing is the most accurate tool to measure and monitor hormone status.

Both men and women experience changes in hormone levels with age.  Sometimes those changes result in unpleasant symptoms that demand attention.   Often, the changes are more subtle - yet there is still an impact on overall health.  Hormone testing is applicable for:

  • Men and women concerned with changing hormone levels as a result of age.
  • Cycling women experiencing PMS symptoms, perhaps related to a hormonal imbalance.
  • Peri- and post-menopausal women concerned with their estradiol and progesterone levels for replacement considerations.
  • Those wishing to monitor their hormone levels following replacement therapy (oral, sublingual or topical), and subsequently regulate their supplement levels.
  • Anyone with symptoms involving fatigue, insomnia, stress, immunity problems, blood sugar problems, and an overweight body should be tested for cortisol levels as well as "sex" hormones.

Men and women of any age who are having symptoms of hormone imbalances should test for all hormones that may be associated with their symptoms. Men and women over the age of forty may want to do a baseline test. Frequently, imbalances will be detectable for a time period before symptoms gain attention.

The major sex hormones to assess are estradiol, progesterone and testosterone. Estrone and estriol are also important sex hormones to consider testing. The main adrenal hormones are DHEA and cortisol. These seven hormones will provide crucial information about deficiencies, excesses and daily patterns, which then result in a specifically tailored treatment approach and one far more beneficial than the old "shotgun" approach. Below is a brief description of each of these hormones:


There are three forms made by the body: estrone, estradiol and estriol. The form used in past hormone replacement therapies is estradiol, often in the form of concentrated pregnant mare’s urine (premarin). It is a proliferative (causes growth) hormone that grows the lining of the uterus. It is also a known cancer-causing hormone: breast and endometrial (uterine) in women and prostate gland in men. It will treat menopausal symptoms like hot flashes, insomnia and memory-loss. With the bio-identical formulas estriol is matched with estradiol (biest) to provide protective effects and additional estrogenic benefits. The other major protector in keeping estradiol from running amok is progesterone. Estrone and estriol are also useful hormones to test.


Called the anti-estrogen because it balances estradiol’s proliferative effects. It is considered preventive for breast and prostate cancers as well as osteoporosis. In addition, too little progesterone promotes depression, irritability, increased inflammation, irregular menses, breast tenderness, urinary frequency and prostate gland enlargement (BPH).


An anabolic hormone (builds tissue) that is essential for men and women. The proper level of testosterone is necessary for bone health, muscle strength, stamina, sex drive and performance, heart function and mental focus.


An important adrenal gland hormone, which is essential for energy production and blood sugar balance. DHEA is a precursor to other hormones, mainly testosterone.


Your waking day hormone (highest in the morning and lowest at night). It is necessary for energy production, blood sugar metabolism, anti-inflammatory effects and stress response.


Classified as a hormone but also acting as a neurotransmitter, melatonin is primarily secreted during the dark (at night) with levels dropping dramatically after exposure to bright light. Melatonin is involved with circadian rhythms, and most commonly associated with sleep patterns and sleep disturbances.

Some of the common imbalances identified through hormone testing include estrogen dominance, estrogen deficiency, progesterone deficiency, androgen (testosterone and DHEA) excesses or deficiencies, adrenal dysfunction and adrenal fatigue.

Pooled Tube Provides More Accurate Sex Hormone Levels

Sex hormone (E2, Pg, T, DHEA) levels fluctuate throughout the day as well as throughout the month while other hormones, like cortisol, have a predictable diurnal rhythm. As a result, many lab measurements will give falsely elevated or depressed values if only evaluating one sampling for sex hormones.

The graph below shows the measured salivary estradiol levels (in pg/ml) of two females during the luteal phase, with samples taken every 10 minutes. As you can see, the estrogen levels are fluctuating continuously and dramatically. Imagine if you had only measured one sampling… say in the morning? It might alter your assessment and lead you to incorrect treatment choices.

Diurnal Changes of Saliva Estradiol in a Femal During Luteal Phase

A Better Reflection of Your Patient’s Hormonal Status

When saliva kits are received by the lab, a fifth tube of saliva is created by pooling a measured sample from each of the four submitted tubes. This fifth pooled tube is mixed thoroughly to provide homogenization and becomes the source from which estradiol, progesterone, testosterone and DHEA are measured. The pooling of these multiple samples throughout the day enables us to provide a much better reflection of each patient’s hormonal status. It is essentially an average of the four submitted samples and more accurately reflects the physiologic hormone levels.

Cortisol Levels Are Measured From Four Timed Samples

Cortisol levels are measured from the original four saliva tubes to provide an assessment of the natural diurnal rhythm of cortisol. Cortisol should be highest in the morning, and gradually taper off throughout the day. Knowing the time of day that a sample was collected is critical to accurate interpretation of the cortisol levels.

Melatonin is Measured From Three Timed Samples

Like cortisol, melatonin is measured directly from the AM, evening and PM sample. Melatonin levels follow a diurnal rhythm with highest levels produced at night, when sleep should be occurring, and the lowest levels should be during daytime.

To read more about the validity of saliva testing you may review these references.
We Cater to the Healthcare Practitioner...
  • Sample collection tubes are small; only 1 ml of saliva is required from the patient. Other laboratories require up to 7.5 ml for the AM sample alone.
  • Fast turnaround times.
  • The four samples collected throughout the day are pooled, giving you an average of the values for estrone, estradiol, estriol, progesterone, testosterone and DHEA. While these hormones do not have diurnal rhythms, their values fluctuate greatly throughout the day. Therefore, an average of these hormones is considerably more relevant than a single AM value.
  • Collection tubes are color coordinated to simplify the timed sample collection process. Additionally, straws are provided for sample collection, which reduces the risk of specimen contamination.
  • Professional/Traditional pricing is available.
  • Labrix offers secure website access for practitioners to retrieve their patient’s results (www.labrix.com or www.doctorsdata.com).
  • Labrix and Doctor's Data work directly with healthcare practitioners only, and will not discuss test results directly with patients. Patients are referred back to the practitioner for consultation and follow-up.
  • Provider education and patient outreach resources are available to help educate your patients about the importance of saliva hormone testing and hormone balancing.