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Why Use Saliva Testing

Why is saliva used for hormone testing?

Saliva is a biological specimen that is easy to obtain, does not require special treatment after collection and can be transported at room temperature. For these simple reasons, saliva has rapidly gained in popularity for diagnostic investigations by medical practitioners but also for use in the home.

The endocrine organs (for example the adrenal gland) produce and secrete hormones into the blood stream that travel around the body to their site of action (for example the ovaries). In the blood however, the hormones are mostly bound to binding proteins which prevent the hormone from activating its receptor at the target tissue. So when a hormone is attached to its binding protein it is biologically inactive.

Measurements of steroid hormones in blood can give a false representation of the level of biologically active hormone because it does not distinguish between bound and unbound fractions. In saliva, only the free, unbound hormone is present, therefore representing the level of active hormone and giving a truer picture of hormone function.

Another key advantage of saliva testing is its practicality for tracking the secretion pattern of hormones that are released in pulses during the day – these hormones should show distinct daily patterns.

Typical examples of pulsatile hormones are the adrenal hormone cortisol, which rises in the morning and falls at night, and the pineal hormone melatonin, which is high at night but low in the morning. TestoChecker provide multiple tubes for convenient sample collection at predefined times of day.

Saliva hormone testing is also recommended for monitoring hormone levels following hormone replacement therapy (HRT) that is administered topically (e.g. in a cream, gel or patch) and can be used to help determine the correct dose of hormone supplementation, in consultation with a medical specialist.

How accurate is hormone saliva testing?

Very! Aside from the practical aspects of saliva hormone testing, the tests are widely accepted in the medical profession to provide an accurate and reliable measure of hormone levels at the time of collection.

Once the saliva specimen arrives at the laboratory, it undergoes rigorous testing that is both sensitive and highly specific to the hormone being tested. The advanced technology can detect very low levels of a given hormone and employs antibody binding methods that lead to only the specific hormone of interest being measured (avoiding false positives). The laboratory applies strict quality control checks ensuring that their test results are high quality and accurate.

The popularity of saliva hormone testing has led to the establishment of well-defined hormone reference ranges in healthy individuals, meaning that hormone results can be compared against the most relevant typical range.  Factors such as sex, age and menopausal status all affect normal hormone ranges, and can be adjusted for, providing an accurate assessment of hormone profile relative to expected norms.

What is the best time of day to use the TestoChecker saliva kit?

Generally, it is recommended to use the saliva testing kit in the morning before eating. It is best to assess hormone levels after a period of fasting; however, it is okay to drink water before the test. Hormone levels tend to peak in the morning after wakening so this is the best time to measure them.

If comparing the level of a specific hormone on different days it is good practice to carry out the sampling at the same time of day each time.

Exceptions are in place for multipoint hormone measurements when the same hormone is measured 2–4 times over the course on one day. For example, an adrenal function test measuring levels of daily cortisol secretion have morning, afternoon and evening timepoints. A cortisol awakening response (CAR) test has 3 timepoints in the morning. A melatonin test has early morning and late-evening collection points.

What forms of androgens do you test?

The body produces multiple forms of androgens that serve important physiological roles in both men and women. At TestoChecker two major types of androgens can be tested:


DHEA-S (dehydroepiandrosterone-sulfate) and DHEA (dehydroepiandrosterone) are androgens that are made in the adrenal glands. The sulfate (-S) group on DHEA helps to stabilise and store DHEA in the blood until it is needed. Measuring DHEA-S in saliva is therefore a reliable measure DHEA production and an indicator of adrenal function in both men and women. DHEA is the precursor of multiple steroid hormones including the sex hormones, estrogen and testosterone. Therefore, DHEA is an important hormone for sexual function, but is also related to bone health, mood, metabolism and maintenance of energy levels.

Testosterone test

Testosterone is made in the testes and the ovaries, from precursors made in the adrenal gland (DHEA). Therefore, poor adrenal function can lead to lowered testosterone production. Testosterone has potent masculinising actions, which is why it is predominantly made in men (but smaller amounts are needed in women). Testosterone influences sex drive, fertility and mood. It also boosts muscle growth and bone strength and contributes to energy and fitness.

What forms of estrogens do you test?

The body produces multiple forms of estrogens that serve important physiological roles in both men and women. Estrogens are made in multiple tissues including the reproductive organs (ovaries and testes), but also liver, fat, adrenal glands and the brain. At TestoChecker three major types of estrogens can be tested:

Estrone (E1) hormone test

Estrone is a weak form of estrogen mainly produced in fat cells and a small amount in the reproductive organs. Estrone is produced in both sexes and can act in its own right or be converted into estradiol (E2). During menopause, when estradiol levels fall sharply, estrone can compensate somewhat for the decline in estradiol.

Estradiol (E2) hormone test

Estradiol is the most potent acting and abundant form of estrogen that is predominantly produced by the ovaries and testes (smaller amounts in men). Estradiol is converted from testosterone and plays an important role in building and maintaining bone mineral density and strength. Estradiol is responsible for fat deposition and has roles in cardiovascular health. In women, estradiol regulates reproductive function including puberty and breast development, and has an essential role in the menstrual cycle, peaking prior to ovulation.

Estriol (E3) hormone test

Estriol is the weakest form of estrogen and has a role during pregnancy. Estriol is mostly made in the placenta and is the most abundant form of estrogen during pregnancy. Estriol may contribute to morning sickness and block the action of other forms of estrogen including estradiol (E2).

When is the best time of month to measure the reproductive hormones?

In men, the reproductive hormones can be measured at any time; however, in women it gets a bit more complicated!

I have regular menstrual cycles

The steroid hormones estradiol and progesterone regulate the menstrual cycle and therefore rise and fall at different phases. A typical menstrual cycle completes every 28 days.

Estradiol levels are extremely variable across the course of the menstrual cycle. Estradiol levels are at their lowest during menstruation (day 1-7) and then increase during the remaining follicular phase (8-14 days), peaking just prior to ovulation at day 14. Estradiol levels fall sharply post-ovulation but peak again during the luteal phase around day 21 (at the same time as the rise in progesterone) before declining once more prior to menstruation. Estradiol tests can be performed throughout the cycle but levels should be referenced against established typical ranges for the specific phase of the cycle in which testing was done.

Progesterone levels peak in the luteal phase following ovulation. A progesterone test should be performed at approximately day 21 of the cycle (21 days after the beginning of the last period) to capture peak secretion of this hormone.

Testosterone (and other androgen) levels do not vary much over the menstrual cycle but may increase slightly around ovulation.

Charts used to help you determine the ideal test date are provided with the TestoChecker Fertility Hormone Balance Test Kits.

I am using hormonal birth control

If using hormonal birth control, such as oral contraceptives, then the sex hormones will be suppressed to prevent ovulation. This means that the timing of hormone testing can occur at any time. If oral contraceptives are ceased it is recommended to wait until the second cycle before testing sex hormone levels.  

I am not cycling due to menopause, hysterectomy or amenorrhea (absence of cycles in reproductive-aged women)

Testing can occur at any time of month. If using topical (applied to the skin) HRT or supplementation, saliva testing should not occur on the same day as this may give false results.

Why measure cortisol levels?

Cortisol is a glucocorticoid hormone produced by the adrenal glands, commonly identified as the body’s ‘stress hormone’.  In times of physical or emotional challenge, cortisol is pumped into the body providing energy, boosting the immune system, controlling blood pressure and regulating metabolism (as well as multiple other actions). We need cortisol to function, but too much or too little cortisol can be detrimental.

A cortisol test can address concerns about adrenal function. In cases where insufficient adrenal hormones are produced, this may indicate adrenal fatigue. Adrenal fatigue can occur if the adrenal gland is overstimulated by factors (including stress, exercise, obesity and trauma) for a very long time.

Cortisol is secreted in a circadian rhythm – it rises in the morning to energise us for the day ahead, and lowers at nighttime as we prepare for rest and sleep. Multipoint comprehensive cortisol tests provide valuable information about overall adrenal function. TestoChecker obtain saliva samples collected in the morning, afternoon, evening and nighttime to generate a comprehensive report on the daily cortisol secretion pattern.

TestoChecker also offer a cortisol awakening response (CAR) test which assesses how the body prepares for the events of the day. Cortisol levels typically rise sharply 30 minutes after awakening and then decline to baseline levels in the next hour. The extent of the CAR can indicate how well the body is primed and ready for the day to come.