ZRT Laboratory (Salivary Steroids)

11-deoxycortisol is an adrenal hormone. It is the end product of 17-hydroxyprogesterone (17OHPg) through 21-hydroxylase synthesis and is the immediate precursor of cortisol.

Levels of these cortisol precursors and the enzymes that stimulate cortisol synthesis from them are typically increased when ACTH levels are increased, which can occur with Cushing syndrome, adrenal carcinoma, ACTH-producing tumors, or 11β-hydroxylase deficiency, a more rare form of CAH34 than seen with 21-hydroxylase deficiency, which constitutes > 95% of all CAH cases

The hormone 17-hydroxyprogesterone is produced by the adrenal glands. 17-OHPg is converted to cortisol, which is released in varying amounts, but at particularly high levels during times of physical or emotional stress. A cortisol deficiency can occur in certain people which can lead to an increase in 17-OH progesterone in the blood. LCMS saliva testing has allowed accurate determination of 17-OH progesterone along with other androgens, correlating with serum levels and allowing useful steroid profiling in disorders of steroid metabolism19. High levels of 17-OHPg can indicate CAH. CAH is a glandular disorder that results in the adrenal glands being unable to create enough cortisol, which may consequently increase the production of DHEA and testosterone.

7-keto DHEA (also known as 7-oxo DHEA) is a steroid produced by metabolism of DHEA.

It is not directly converted to testosterone or estrogen.

7-keto DHEA is rapidly absorbed when given as a supplement and converted to its sulfate derivative. It is commonly used to produce the metabolic effects of DHEA while avoiding metabolism into estrogens or androgens, and clinical research supports its role in benefiting metabolism and weight management. Endogenous 7-keto DHEA may have some anticortisol activity through enzyme competition which in the case of hypercortisolism may be beneficial to the adverse effects of cortisol on metabolic syndrome. Most studies on 7-keto DHEA are on improving the metabolic rate where there appears to be improvement in metabolism despite being on a low caloric diet. There is also limited information that 7-keto DHEA may act to increase levels of T3 while patients are on a caloric restricted diet.

Aldosterone is a mineralcoritcoid and a hormone. It allows the transport of sodium across the cell membrane. This is especially important in the kidney (distal tubule). Because of its function, aldosterone is important in blood pressure regulation and also for the volume of blood found in the blood vessels. Potassium is an antagonist to sodium. If potassium is high, sodium levels will be low. When potassium is found high in the plasma of the blood, the adrenals step in and synthesize aldosterone. The synthesis of aldosterone is taken care of in the body primarily by the renin-angiotensin system.

Allopregnanolone, also known as brexanolone, is a medication and a naturally produced steroid that acts on the brain. Allopregnanolone possesses a wide variety of effects, including, in no particular order, antidepressant, anxiolytic, stress-reducing, rewarding, prosocial, antiaggressive, prosexual, sedative, pro-sleep, cognitive, memory-impairment, analgesic, anesthetic, anticonvulsant, neuroprotective, and neurogenic effects.

Anastrozole is a medication that inhibits the enzyme aromatase to suppress testosterone conversion to estrogens.

Anastrozole is used in combination with other treatments for suppressing testosterone conversion to estrogens. It can be used in combination with other treatments, typically men using testosterone therapy to prevent conversion to estrogens; and in breast cancer and prostate cancer patients to inhibit endogenous estrogen production that could stimulate estrogen-sensitive tumor growth. It is most often used for hormone-receptive breast cancer.

It works by binding to the aromatase enzyme and blocking the conversion of androgens to estrogens in peripheral tissues. Off-label it is commonly used to decrease the production of estrogen in men and is also used as part of a treatment plan for women with endometriosis.

Androstenedione is secreted predominantly by the adrenal gland and production is controlled, in part, by adrenocorticotropic hormone (ACTH). It is also produced in the testes and ovaries from DHEA-S. It is a weak androgen and an intermediate in the biosynthesis of testosterone and estrone from DHEA. It has been found to have some estrogenic activity.

Androstenedione is converted to estrone by the action of aromatase in fat tissue.

Corticosterone, also known as 17-deoxycortisol, is a steroid hormone of the corticosteroid type produced in the cortex of the adrenal glands. Corticosterone has multiple effects on memory. The main effects are seen through the impact of stress on emotional memories as well as long term memory. With emotional memories, corticosterone is largely associated with fear memory recognition. Not only does corticosterone have effects on emotional memories but memory recognition and consolidation as well.

Under the direction of the hypothalamus and pituitary, and controlled by a negative feedback loop, the zona fasciculata of the adrenal cortex is stimulated to produce cortisol in response to circadian peaks and troughs in ACTH synthesis in addition to various types of stressors such as emotional/psychological, physical (injury, exercise), chemical, pathological (viruses, bacteria, etc.).

The feedback loop is commonly referred to as the hypothalamic-pituitary-adrenal (HPA) axis.

In a normal nonstressed state cortisol production is at its highest upon waking and declines steadily during the day, reaching its lowest point at bedtime.

Salivary cortisone is an inert form of cortisol, as is estrone to estradiol.

Simultaneous testing for cortisol and cortisone assists in diagnosing acquired or inherited abnormalities of 11β-hydroxy steroid dehydrogenase, affecting the cortisol to cortisone ratio.

Deficiency of 11β-HSD results in a state of mineralocorticoid excess because cortisol, not cortisone, acts as a mineralocorticoid receptor agonist.

Dehydroepiandrosterone (DHEA), a hormone produced by the adrenal glands, is the precursor for estrogens and testosterone, and is therefore normally present in significantly greater quantities than all the other steroid hormones. It is mostly found in the circulation in the form of its sulfate ester, DHEA sulfate (DHEA-S), levels of which in saliva are higher and more stable than those of DHEA. Its production is highest in the late teens to early 20s and declines gradually with age in both men and women.

Dehydroepiandrosterone (DHEA), a hormone produced by the adrenal glands, is the precursor for estrogens and testosterone, and is therefore normally present in significantly greater quantities than all the other steroid hormones. It is mostly found in the circulation in the form of its sulfate ester, DHEA sulfate (DHEA-S), levels of which in saliva are higher and more stable than those of DHEA. Its production is highest in the late teens to early 20s and declines gradually with age in both men and women.

Levels of DHEA-S reflect adrenal gland function.

Dehydroepiandrosterone (DHEA), a hormone produced by the adrenal glands, is the precursor for estrogens and testosterone, and is therefore normally present in significantly greater quantities than all the other steroid hormones.

Dihydrotestosterone is an endogenous androgen that is formed from testosterone via 5α-reductase activity in certain tissues including the prostate gland, seminal vesicles, epididymis, skin, hair follicles, liver, and brain.

DHT, relative to testosterone, is more potent as an agonist of the androgen receptor. Inhibition of 5α-reductase activity to reduce prostatic DHT levels is used to treat benign prostatic hyperplasia (BPH).

DHT has been used clinically as treatment for low testosterone levels in men. DHT is biologically important for sexual differentiation of the male genitalia during embryogenesis.

Circulating levels of DHT are low in relation to testosterone. Deficiency in 5α-reductase results in incompletely virilized males which is clinically supported by an elevated ratio of testosterone to DHT.

Estradiol is the predominant, and the most potent, circulating estrogen. Bioavailable estradiol, which represents about 2% of the total protein-bound estradiol in the bloodstream , exits the bloodstream in capillary beds and enters target cells such as the brain, breasts, uterus, bone and heart.

Estriol is the weakest of the three major naturally-occurring estrogens in women.

Estriol is a product of the metabolism of estrone and estradiol. Because of its weak estrogenic activity, estriol is sometimes preferred for intravaginal use as an alternative to systemic estrogen therapy for the treatment of urogenital atrophy in postmenopausal women.

It is also used in anti-aging skin creams as a form of topical estrogen replacement to counteract the effects of age-related estrogen loss on skin. Estriol is the major estrogen found in the maternal circulation during pregnancy; 90% of this circulating estriol is the product of metabolism of DHEA from the fetal adrenals, and so maternal estriol levels are used as an indicator of fetal health.

In non-pregnant women, estriol levels are similar in both pre- and post-menopause and are also similar to levels in men. Salivary estriol has been found to be predictive of increased risk of preterm labor in pregnant women. In nonpregnant women, it is most commonly used for monitoring of levels in women using estriol-containing supplements as part of hormone replacement therapy.

Produced by the ovaries, the estrone hormone is one of three types of estrogen, and it is one of the major hormones found in the bodies of postmenopausal women. While research into estrone function is still ongoing, since it is the least powerful of the three estrogen types, women should still understand this hormone and its known effects on the body.

- Weaker compared to Estradiol (Research says the estrogenic activity is about 4% of estradiol’s activity)

- Most abundant in menopause

- Made via aromatization in several tissues like fat and muscle

- Converts into estradiol (E2)

Estrone (E1) is also made by the ovary but in fat tissue in lesser quantities. While not as abundant in circulation as estradiol, estrone excess can still increase the risk for estrogen dominant cancers and estrogen dominant symptoms such as breast tenderness, heavy menstrual cycles, headaches, and erectile dysfunction and breast development in men just like estradiol. Estrone is commonly thought to be more abundant during menopause. 

Estradiol and estrone can interconvert into each other. 

Ethinyl estradiol is an estrogen receptor agonist commonly used in combined oral contraceptives. It is a synthetic derivative of estradiol. In the liver it stimulates the synthesis of SHBG, increasing SHBG levels by 2- to 4-fold in women, which has the effect of binding more circulating testosterone and reducing free testosterone concentrations by 40-80%. The suppression of free testosterone levels may impact female sexual desire.

Finasteride is a 5α-reductase inhibitor used to block the formation of the potent androgen DHT from its precursor, testosterone. It is used to treat benign prostatic hyperplasia as well as male pattern baldness in men and women, and excessive facial or body hair growth in women.

Serum testosterone levels increase as a result of the reduced conversion to DHT, but the increase is not usually outside the normal range

. Treatment with finasteride has been linked with sexual side effects.

Letrozole is an aromatase inhibitor used as an adjuvant treatment for hormone-dependent breast cancer. It inhibits peripheral estrogen production in fat tissues, where it prevents the conversion of testosterone into estradiol. It has similar safety and efficacy to anastrozole.

Melatonin is a hormone that is produced from the pineal gland in a circadian pattern and plays a role in the initiation of sleep.

The production and release of this hormone is connected to the time of day, ideally increasing when it is dark and decreasing when it is light. Melatonin and cortisol follow opposite circadian patterns but are not cross-regulated in a negative feedback manner.

The production of melatonin decreases with age.

Treatment with melatonin may be useful in people with circadian rhythm sleep disorders, such as delayed sleep phase disorder, jet lag, shift worker disorder, and the non-24-hour sleep-wake disorder most commonly found in totally blind individuals.

Pregnenolone sulfate is a neurosteroid that enhances the glutamate N-methyl-D-aspartate (NMDA)-receptor function and inhibits receptors for glycine, GABA, thereby regulating the excitation-inhibition balance in the CNS.

It enhances learning and memory and promotes nerve cell survival. Along with other endogenous neurosteroids, it has been implicated in the development of neuropsychiatric disorders such as schizophrenia, depression, and anxiety. Trials of the therapeutic use of pregnenolone, the precursor of pregnenolone sulfate, in schizophrenia have shown some success. Low plasma levels of pregnenolone sulfate have been observed in people with generalized social phobia.

Progesterone’s primary function during the menstrual cycle is to induce a secretory endometrium ready for implantation of a fertilized egg. Levels therefore increase during the luteal phase of the cycle after ovulation. If no implantation occurs, progesterone returns to follicular phase levels.

Progesterone’s primary function during the menstrual cycle is to induce a secretory endometrium ready for implantation of a fertilized egg.

Levels therefore increase during the luteal phase of the cycle after ovulation.

If no implantation occurs, progesterone returns to follicular phase levels. If a pregnancy results, progesterone continues to rise to very high levels and carries out a variety of functions necessary to sustain the pregnancy.

In some patients with infertility, ovulation may occur but luteal phase levels of progesterone are inadequate. Luteal phase deficiency is a result of inadequate progesterone production by the corpus luteum.

During menopause, ovarian progesterone production dwindles, resulting in postmenopausal levels similar to those seen in men.

Progesterone has wide-ranging physiological effects, including neuroprotection, maintenance of skin elasticity, and development of bone tissue. Progesterone also counteracts the proliferative effects of estrogen on the endometrium. When samples are collected after transdermal application of progesterone, saliva progesterone levels are higher than serum, indicating distribution of progesterone to tissues.

Optimal: 100-500 when E2 1.3-3.3 pg/mL

This ratio is helpful when both E2 and Pg are within range, yet the patient continues to have symptoms. It is not expected to be normal or used clinically when either E2 and/or Pg are outside of their expected ranges or if the patient does not have clinical symptoms.

Is the ratio relevant in women using hormone therapy?

With some types of hormone therapy such as topical progesterone, Pg levels in saliva are much higher than endogenous luteal phase levels, ranging from 200-3000 pg/mL at 12-24 hours after dosing, and so the ratio can appear high. However, because symptoms of both estrogen dominance and progesterone dominance can look the same, testing and assessing the ratio along with clinical symptoms can help determine the next step for treatment.    

In men, levels of testosterone begin to decline with age, usually beginning around the mid-40s. The decline in testosterone production by the testes can be more precipitous in some men than others.

Excessive weight gain, stress, lack of exercise, and many medications can further reduce testosterone levels, leading to symptoms that include low libido, irritability, depression, loss of muscle mass and strength, weight gain, erectile dysfunction, osteoporosis, and adverse changes in the blood lipid profile.

Testosterone levels in saliva are an accepted method for assessment of hypogonadism in men.

In women, high testosterone, often caused by ovarian cysts, leads to conditions such as excessive facial and body hair, acne, and oily skin and hair.

Low testosterone in postmenopausal women, seen particularly after surgical removal of the ovaries, leads to female symptoms of androgen deficiency including loss of libido, thinning skin, vaginal dryness, and loss of bone and muscle mass.

Testosterone levels in saliva are an accepted method for assessment of hypogonadism in men. In women, high testosterone, often caused by ovarian cysts, leads to conditions such as excessive facial and body hair, acne, and oily skin and hair. Low testosterone in postmenopausal women, seen particularly after surgical removal of the ovaries, leads to female symptoms of androgen deficiency including loss of libido, thinning skin, vaginal dryness, and loss of bone and muscle mass.