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OptiMale
Testosterone Health Check Kit
1 Kit

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Assess Your Testosterone And Other Important Male Hormones!

Based on more than 20 years of research, salivary hormone assessments have become a preferred method of measuring hormonal health. In addition to being less expensive and more practical than blood testing, salivary assessments measure the active (bioavailable) levels of hormones rather than the inactive (total or bound) levels of hormones.

Did you know?

  • Low testosterone and DHT levels can greatly effect energy, mood, physical strength and sexual function.
  • Modern lifestyle along with chemical pollution can cause elevated levels of male estrogen which can reduce libido, increase body fat and enhance your risk of cardiovascular disease and prostate disease.
  • The proper balance of DHEA and cortisol make up the bodyÆs inner defense against the pressures of stress, helping your body adapt in a healthy manner.

How is your testosterone health?

  • Are you over the age of 35?
  • Are you under a lot of stress?
  • Do you feel fatigued?
  • Are your muscles sore for a long time after a workout?
  • Have you noticed a waning sex drive?
  • Have you gained weight, particularly around your mid-section?
  • Do you feel depressed, irritable or unmotivated?
If you answered yes to two or more of these questions, you could be suffering from male hormonal imbalance.

Testosterone Health Check Details:

Before You Start:
To ensure the accuracy of your health check, please read all the instructions carefully and ensure all packing materials are present before collecting your sample.

Materials in Box:
1 collection tube with time label for saliva, 1 parafilm square to aid in saliva collection, 1 specimen time/date label, 1 resealable plastic bag, 1 absorbent pad, 1 USPS mailing label, 1 priority mail sticker, 1 return shipping box, 1 hormone assessment request form.

When to Collect Sample:
Collect your saliva sample within one hour after rising between 6 AM and 9 AM. Prior to collection, it is imperative that you keep anything out of your mouth other than water.
DO NOT EAT, DRINK, BRUSH TEETH, USE MOUTHWASH OR TAKE MEDICATIONS OR SUPPLEMENTS!

Collection Instructions:
1. Drink at least 16 ounces of water. Rinse and swish 3 times for 5 seconds with cold water. Wait 5 minutes before collecting. Do NOT drink once you've started.
2. In the tube provided, collect your saliva until 3/4 full of liquid, NOT foam. Most people take about 15 minutes to complete. In order to remove any foam, tap the covered tube on a hard surface to eliminate bubbles. If you find that you are having difficulty producing enough saliva, chew the clear parafilm while collecting. Discard the parafilm when done.
3. Tightly replace the cap to prevent leakage. On the specimen label, write your name, circle saliva and record the date and time of collection. Wrap the tube in the absorbent pad and place inside the enclosed resealable bag. FREEZE the sample for at least 3 hours.
4. Please the sealed bag along with your completed Hormone Assessment Request Form along and any addition funds in the box provided. Check saliva on the pre-paid USPS label and use it to seal the box. Apply the mailing sticker to the box. If you desire faster results, please ship sample via expedited carrier to the address on the label.

Results:
The results of you assessment will be mailed to you by the lab within 5 days of receipt.

About Salivary Testing

The knowledge that there are steroid hormones in saliva, and that they can be measured, has been around for more than 30 years. But it is only recently that technology has caught up with that information making it possible to accurately determine levels of salivary hormones. Steroid hormone levels have and still are most commonly carried out on blood samples. But blood collection has its limitations. It is invasive, expensive and difficult to time precisely.

Perhaps more importantly, blood hormone levels represent the body's total hormone content. Since most blood hormones (about 95%) are bound to specific proteins which carry them through the bloodstream they can be considered the hormone 'storage' fraction. The other 5% represent the free hormones - those available to move easily into the target organs and fulfill their functions. Saliva contains these free hormones which can be easily measured to give an accurate picture of those hormones that are readily available to the tissues.

Many studies in the scientific literature have shown that there is a strong correlation between the levels of steroid hormones in saliva and the bio-available (free) levels of steroids in the bloodstream. The amount of steroid hormone that enters the salivary ducts and then saliva, is representative of the fraction of steroids in the bloodstream that are bio-available to other tissues in the body.

The Advantages of Saliva Testing
Saliva collection is easy, can be done anywhere, any time and very inexpensive. Serum hormone testing can be stressful and inconvenient. Blood testing must be scheduled and it is more difficult to obtain samples at the desired times (e.g. early am) and multiple times through the day or late evening. There is also an extra cost involved in drawing blood, independent of the cost for the test itself.

Hormones in saliva are exceptionally stable, allowing wide latitude in collection and shipment. Samples can be stored at room temperature for at least a week without loss of activity, so samples can be shipped to the testing facility by regular mail. Blood, on the other hand, must be kept cool on ice packs, increasing costs for shipping as well as the likelihood of error due to improper handling.

Collection and Processing
Five milliliters of saliva is requested. Following collection, saliva should be either frozen or shipped to the lab within one day of collection. Saliva is stable at ambient temperature for at least three weeks and a specimen received within that time is acceptable.

Test results are then generated and mailed within five to seven working days from date of receipt of the specimen to the laboratory, often sooner.

How Can Saliva Hormone Tests Help Your Patient?
The goal of saliva hormone monitoring is to determine whether or not hormone replacement therapy or another form of intervention is appropriate. If your patient is already taking hormones, testing will allow you to adjust dosages, both optimizing benefits and minimizing risks. And since your patient may opt to receive a copy of the test results directly, they will be able to play a more active role in their own health care and are more likely to adhere to the proposed hormone replacement intervention.

Why Test for Hormones?
  • To establish a baseline before entering into a program of hormone supplementation
  • To establish if there is a deficiency of hormones or an imbalance of hormones
  • Confirmation that hormone supplementation is working
  • To help confirm a medical diagnosis that may be hormone related

Advantage of Saliva vs. Serum Testing

Closely Represents Physiological Levels

  • Saliva reflects the biologically active (free) fraction of hormones in the blood stream (unlike blood or urine, which measures the total levels)
  • Saliva testing allows for practical collection of multiple samples to assess the circadian (cyclical) rhythm of hormones such as testosterone and cortisol.
Patient Friendly

  • Saliva testing is non-invasive, simple, stress free and painless. NO NEEDLES!
  • Saliva can be convenient collected in the privacy and comfort of home.
  • Saliva testing is less expensive than blood testing.
  • Because hormones are stable in saliva for two weeks or more samples can be sent to the laboratory by regular mail without special handling
More Advantageous to the Doctor

  • Specimen collection supplies can be provided to the practice in a timely manner.
  • Technical and clinical support from MedLean
  • Easy-to-read test results, graphs and ranges
  • Rapid turnaround time of patient results
  • Comprehensive professional guide for test result interpretation and technical information
  • Availability of patient educational material

Comparison of Salivary Testosterone versus Serum Free and Total Testosterone Levels in Men with Symptoms of Low Testosterone

Subject Symptom Saliva Free Testosterone Total Testosterone
97-478 pmol/L 35-195 pg/ml 200-835 ng/dl
34 year old power lifter Irritable, fatigued, joint pain, sleep difficulties 246 137 465
57 year old teacher Low energy, reduced sex drive and weight gain. 205 126 499
45 year old business man Fatigue, apathy, loss of muscle, low sex drive 163 84 443
40 year old professional skater Apathy, fatigue, loss of stamina, agility and strength 150 XX 309
52 year old computer executive Feeling older and less robust, lower libido, mentally sluggish, loss of assertiveness. 93 15 778
54 year old mechanic; history of anabolic steroid use Sensitivity to DHEA, mild gynecomastia, prostate enlargement 38 10 818


All of these men had significant relief of their symptoms after therapy that restored high normal levels of free testosterone.

Ranges of Testosterone in Saliva and Serum

pg/ml GSDL Aeron DiagnosTech Smith Kline Free Test. SmithKline Total Test. ng/dl
Men 20-135 30-140 30-150 35-195 200-825
Woman 10-40 15-40 10-35 1-21 <60


Interpretation of Saliva Hormone Ranges
Laboratory values can only be fully interpreted in the context of the complete clinical picture. Specific diagnoses cannot be made on the basis of laboratory values alone. In addition, for diurnally variable hormones or supplementation, the timing of collection of the specimen is critical. Normal ranges most be calibrated relative to a specific time of day (early AM) and to specific times relative to dosing of supplementation (trough levels). The normal ranges may not be valid for different collection regimens (i.e. PM collection or to capture peak levels).

Derivation of Ranges
Ranges are determined by testing both volunteers representing designated populations and drawing from an extensive patient database. Each range is based on as many as 800 analyses. These salivary hormone ranges capture 90% of normal female and male values leaving 5% below and 5% above the normal range. Correlation of these ranges with those in the literature for saliva steroid hormones is excellent.

Unsupplemented salivary hormone ranges represent normal physiologic levels for the groups of patients described (i.e. premenopausal, postmenopausal, specific age groups and gender for androgens). These ranges do not necessarily represent optimal levels for health since physiologically normal levels can be risk factors for pathologic processes in certain groups (postmenopausal low levels of estrogen put women at risk for bone fracture due to increased bone turnover).

Age-based testosterone hormone ranges are guidelines for targeting levels that give and sustain optimal health. Ranges normally given for older populations are representative of normal, unsupplemented hormone levels and not necessarily those which will provide maximum well being. Titration to more youthful target ranges is an important therapy issue to consider.

Supplemented salivary hormone ranges are derived from groups of people on usual dose ranges of standard medications. Thus, these ranges are expected rather than optimal ranges as delineated by clinical endpoints. Dose levels tend to be high rather than low and physiologic effect (i.e. relief of symptoms, decrease in bone turnover rates) can be expected from the lower end of expected dose ranges for supplementation in most patients.

Individual and Physiological Variability
Some ranges may seem broad but this variation aptly demonstrates the wide individuality of hormone levels. As to be expected, women have higher levels of the estrogens and progesterone than men, and cycling women have higher levels than menopausal women. Men, in general, have higher DHEA and testosterone levels than women, and in both sexes, the level of each drops dramatically with age. Healthy state cortisol levels remain constant throughout life, regardless of sex or age.

Time of Collection
The ranges represent saliva hormone levels from specimens collected in the early morning unless otherwise noted. Since testosterone, DHEA and cortisol express diurnal variation with levels highest just after waking, it is extremely important that patients indicate the time of collection for an effective hormonal evaluation. Saliva hormone ranges were developed with early AM collections (between 6-8 AM), and they may not be valid for collections taken outside this window of time. For patients taking supplementation, there is obvious pharmacokinetic variation relative to the time of dosing. If peak levels are desired, one may adjust the timing of collection to coincide with peak supplemented hormonal levels. However, Clinical Laboratory ranges are not adjusted to interpret peak levels.

Hormone Replacement Therapy (HRT) Pharmacokinetics

Delivery Systems

Oral HRT: Taken as tablets or capsules, oral HRT levels follow a cyclic pattern following ingestion. Oral doses, on average, result in peak levels about 2-6 hours following dosage, falling to the lowest, trough levels between 8-12 hrs.

Transdermal HRT: Hormones can be delivered transdermally or transmucosally (mouth, vagina or rectum) with a cream or gel. Saliva levels of hormones delivered transdermally (such as progesterone which is readily available over the counter in creams) are high relative to physiologic levels because of the following (see full explanation below):

Hormones are more fully absorbed through the skin than from the gut and do not experience the 'first liver pass' which breaks down over 80% of orally delivered hormones. Hormone-containing creams are applied in large amounts (as opposed to patches) and enter the system in a very short time.

When hormones are applied to the skin, we see very significant increases in salivary levels of that hormone, but often we see very little change in the serum level. This is because they are already in a biologically active form and quickly carried throughout the body by the blood’s fatty components such as chylomicrons or red blood cell membranes. They can be detected through the saliva within a few hours of applying the cream. Serum testing is a valid way to test hormones if they are produced within the body endogenously, or if you were taking hormones orally. However when hormones are applied to the surface of the skin, there is very little increase in serum levels but there's a remarkable increase in saliva levels. We believe the reason for this is that when hormones enter through the skin and enter the bloodstream, they're carried by red blood cells to target sites where they're delivered. When blood is collected, the red blood cells are spun down and removed, thrown away, and hormones tested in serum. Therefore, serum is not representative of the bioavailable fractions of hormones in the bloodstream in transdermal delivery. When using a transdermal cream, it's best to not use a cream for 12 to 24 hours before collecting the saliva specimen.

Patch HRT: HRT patches provide a continuous, steady-state delivery and are applied every 3-7 days for woman and daily for men. Patch-delivered hormones usually reach a peak by 24 hours followed by continuous levels for the duration of the recommended patch interval.

They deliver very small amounts of hormones slowly, thus do not overwhelm the binding capacity of the binding globulins. As expected, patches do not show a transdermal cream pattern but give levels that look like endogenous production or oral replacement at about 2 % of serum values.

Sublingual HRT: Sublingual delivery is similar to transdermal and the above explanation applies. However, caution during sample collection must be taken since occasionally the mouth will not clear the preparation and the saliva will be directly contaminated with hormone. If this is a problem, a PM collection prior to the next dose allows the mouth to clear during the day with increased oral activity. This collection modification will solve the problem in most cases.

Scientific Abstracts References

1) Testosterone Concentrations in Human Seminal Plasma and Saliva and its Correlation with Non-Protein-Bound and Total Testosterone Levels in Serum; Sannikka TH, Terho P, Suominen J, et al. Int J Androl 6: 319-330, 1983

A sensitive, specific and precise non-chromatographic method for the radioimmunoassay of testosterone in human seminal plasma and saliva from adult and pubertal males is described, and the values compared to total and non-protein-bound testosterone levels in serum. There was a significant correlation between salivary and serum-free levels of testosterone (r=0.75, P<0.001, n=67) whilst the correlation of serum levels of total testosterone with free as well as with salivary testosterone levels was weaker (r=0.63 and 0.64, respectively). The salivary and serum levels of free testosterone showed better correlation with the stage of puberty than did the serum levels of total testosterone. Further evidence for a correlation between salivary and serum levels of free testosterone was obtained following oral administration of testosterone undecanoate, at this treatment increased the mean concentration of serum total testosterone after 3 h by 82%, but increased salivary and serum levels of free testosterone by only 30% and 20%, respectively. The coefficient of correlation between serum levels of total testosterone and seminal plasma testosterone was 0.73 (P<0.001), whilst the correlation between levels of serum-free testosterone with both salivary and seminal plasma levels of testosterone was statistically non-significant. Our observations on salivary testosterone are in accordance with the diffusion of non-protein-bound steroids into peripheral tissues, and consequently into their secretions. This model, however, does not appear to be applicable to the sex accessory glands.

2) Reliability of Salivary Testosterone Measurements: A Multicenter Evaluation; Dabbs JM, et al. Clin Chem 41: 1581, 1995

The reliability of salivary testosterone assays was evaluated by nine laboratories in four countries. Each laboratory used its own RIA procedures to assay samples from a set of 100 male and 100 female subjects. Agreement among the laboratories on mean scores was within the range reported by Read (Ann NY Acad Sci 1993;694:161-76). Overall agreement on individual scores, as indicated by the intraclass correlation coefficient computed within subjects across laboratories, was r=0.87 for men and r=0.78 for women. Mean agreement between each laboratory and the combined set of all other laboratories (via Fisher’s Z-transformation) was r=0.61 for men and r=0.58 for women. We take these latter values to be the best estimates of the average reliability of laboratories in their ordering of individual samples.

3) Salivary Testosterone Measurements: Reliability Across Hours, Days, and Weeks; Dabbs JM Jr. Physiology and Behavior 48: 83-86, 1990

Salivary testosterone measurements would appear to be useful in behavioral research, where subjects are often reluctant to provide serum samples. The usefulness of salivary measurements depends upon their reliability, however, which was the focus of the present investigation. In four studies, 270 male and 175 female subjects collected saliva samples at times ranging from 30 min to 8 weeks apart. Subjects collected samples on at least two days, at time of awakening, midmorning, late afternoon, and late evening. Mean reliability was r=.64 across two days and r=.52 across seven-eight weeks. Menstrual cycle effects were negligible. Reliability can be increased by using more than one measurement, and it is probably desirable to combine measurements taken several weeks apart. Salivary assays offer a practical way of measuring testosterone in free-ranging subjects outside the laboratory.

4) Direct Radioimmunoassay (RIA) of Salivary Testosterone: Correlation with Free and Total Serum Testosterone; Vittek J, L’Hommedieu DG, Gordon GG, et al. Life Sci 37: 711-716, 1985

Simple and sensitive direct RIA for determination of salivary testosterone was developed by using RSL NOSOLVEX TM 9125 1) kit produced by Radioassay System Laboratories (Carson, California). In addition, a relationship between salivary and serum free and total testosterone concentrations was studied in randomly selected 45 healthy subjects, 5 females on oral contraceptive pills and 28 hypertensive patients on various treatment regimens. The lowest weight of testosterone detectable by our modified method was equivalent to 1 pg/ml of saliva, taking into account analytical variability. Intra- and interassay coefficients of variation were 5.09+/-2.7% and 8.2+/-5.9%, respectively.

Statistically significant correlations were found between salivary and serum free testosterone (r=0.97) and salivary and serum total testosterone concentrations (r=0.70-0.87). The exception to this was a group of hypertensive females in which no correlation (r=0.14) between salivary and total serum testosterone was found. It is also of interest that, while salivary testosterone was significantly increased in subjects taking oral contraceptives and most of the hypertensive patients, the total serum testosterone concentrations was in normal range. Our findings suggest that determination of salivary testosterone is a reliable method to detect changes in the concentration of available biologically active hormone in the circulation.

5) Salivary Testosterone in Men: Further Evidence of a Direct Correlation with Free Serum Testosterone; Wang C, Plymate S, Nieschlag E, et al. J Clin Endocrinol Metab 53: 1021-1024, 1981 An excellent correlation was found between salivary testosterone (T) and serum T concentrations, as measured by RIA. Using polyacrylamide gel electrophoresis, we have demonstrated that sex steroid-binding globulin could not be identified in the saliva of men with serum sex steroid-binding globulin. After exogenous T administration, saliva and serum T rose abruptly and in parallel. Salivary T concentrations in male patients with thryotoxicosis were similar to those in normal males, whereas the serum T and sex steroid-binding globulin values were significantly higher in the hyperthyroid patients. This study demonstrates that salivary T levels may be used a an index of free serum T.

6) Bioavailable Testosterone in Salivary Glands; Pardridge W and Demers LM. Clin Chem 37: 139-140, 1991

The determination of testosterone in the clinical evaluation of hyper- and hypoandrogenic states has historically involved the measurement of total and free testosterone in the circulation. The free testosterone has been determined as the absolute free concentration, which normally represents only 2-3% of the total testosterone. Testosterone circulates bound primarily to sex-hormone-binding globulin (SHBG) and albumin, with a only a very small and insignificant fraction bound to cortisol-binding globulin (CBG). Under normal circumstances, ~44% of the testosterone in the male is SHBG-bound, while 54% is bound to albumin. In females, the figures are 78% SHBG-bound and 20% albumin-bound. In recent years the concept of bioavailable testosterone has emerges, in which the testosterone fraction bound to albumin is considered to be as readily available to the tissues as is absolute free testosterone; the non-SHBG-bound fraction, which includes both the free and albumin-bound fraction, appears to represent the biologically active form of the hormone. A few laboratories have also used the term "free and weakly bound" to indicate the non-SHBG-bound fraction as the free fraction along with the total testosterone concentration.

Recent evidence suggests that steroid measurements in saliva may be a useful, noninvasive approach to the assessment of free hormone concentrations. The salivary concentrate represents a filtered fraction of the hormone and is independent of flow rate. The study by Swinkels et al. (1) in this issue addresses the bioavailable testosterone concept in saliva and examines the issue of non-SHBE-bound testosterone vs absolute free testosterone in this context.

7) Comparison of the Testosterone-to-Cortisol Ratio Values Obtained from Hormonal Assays in Saliva and Serum; Obminski Z. J Sports Med Phys Fitness 37: 50-55, 1997

Testosterone (T) and cortisol (C) were determined in serum and saliva, sampled simultaneously, from triathletes and karate athletes, in order to determine the T:C ratios in those body fluids and the relationship between them, as well as to assess the salivary T:C ratio as a measure of the so-called anabolic-catabolic index. Mean salivary T:C (value (1.67+/-0.85) was nearly 3-fold lower than that obtained for serum (4.87+/-1.86). Salivary and serum values were strongly correlated with one another (r=0.874, P<0.001) but the relationship depended on the range of cortisol concentrations in serum, the slope of the saliva - serum regression line being significantly lower for serum cortisol concentrations over 600 nmol/l than for concentrations below that value (0.305 and 0.380, P<0.05, respectively). It has been concluded that the salivary T:C ratio, based on values reflecting the levels of biologically active fractions of T and C in circulation, is a better measure of metabolic equilibrium conditioned by those hormones that the corresponding ratio obtained from total concentrations in serum.

8) The Diagnostic Uses of Saliva; Mandel ID. J Oral Pathol 19: 119-125, 1990

It is becoming increasingly apparent to investigators and clinicians in a variety of disciplines that saliva has many diagnostic uses and is especially valuable in the young, the old and infirm and in large scale screening and epidemiologic studies. The highly sensitive test procedures that are now commonplace makes it practical to quantitate, despite very low concentrations, a large number of hormones and drugs in saliva. Indeed, all steroids of diagnostic significance in routine clinical endocrinology can now be readily measured in saliva. Drug monitoring can include abusive as well as therapeutic agents. The concordance between anti HIV antibodies in saliva and serum has stimulated application to various other antiviral antibodies as well as to viral antigens per se. Saliva has found use as a diagnostic aid in an increasing number of clinical situations and in systemic diseases that can affect salivary gland function and composition such as Sjögren’s syndrome, cystic fibrosis and diseases of the adrenal cortex. The list keeps growing.

9) Saliva as a Diagnostic Fluid; Malamud D. Br Med J 305: 207-208, 1992

Interest has been increasing in non-invasive diagnostic testing. Some of this attention stems from the AIDS epidemic, which has provided a new rationale for haemophobia, while other factors include new developments in home based diagnostic tests, a demand for samples collected in the home or workplace, and the close linkage of biotechnology to diagnostic tests.

10) Direct Solid-Phase Enzymoimmunoassay of Testosterone Saliva; Howard K, Kane N, Madden A, et al. Clin Chem 35: 2044-2047, 1989

This competitive, solid-phase enzymeimmunoassay for testosterone in saliva is carried out on microtiter plates and involves no chromatographic or extraction steps. With an overnight incubation the detection limit of the assay is 230 fg per well (16.1 pmol/L). There was a good correlation (correlation coefficient 0.95) between testosterone concentrations measured with and without prior extraction of saliva samples containing a range of testosterone concentrations (200-1000 pmol/L) gave within- and between-assay coefficients of variation of 5.5-13.2%. The analytical procedure is simple and closely resembles already published procedures for the determination of progesterone and estrone (with extraction) in saliva. One person can assay 200 samples in 24 h and the assay is suitable for reproductive and sports medical application, particularly for projects involving serial sampling and yielding large numbers of samples.

11) Menstrual Variation in Salivary Testosterone Among Regularly Cycling Women; Campbell BC and Ellison PT. Horm Res 37: 132-136, 1992

To determine menstrual variation in salivary testosterone daily saliva samples were collected from 20 regularly cycling women. Results indicate that the menstrual profile of salivary testosterone for both ovulatory and anovulatory cycles exhibits local peaks during the follicular phase and at midcycle, as well as a luteal trough. However, the testosterone profile for anovulatory cycles exhibited a later midcycle peak than that for ovulatory cycles, as well as significantly higher average testosterone levels. These results extend the observation of a midcycle peak in serum testosterone to saliva and suggest the existence of a follicular peak in unbound testosterone coincident with the early androgen production of a cohort of developing follicles.

12) Hormones in Saliva: Mode of Entry and Consequent Implication for Clinical Interpretation; Vining RF, McGinley RA and Symon RG. Clin Chem 29: 1752-1756, 1983

Assay of hormones in saliva would be more convenient than assay in blood, but there is no information on the route by which hormones enter saliva, information that would provide insight into the clinical value of such assays. We have examined the mode of entry of various hormones into saliva. The results suggest that unconjugated steroid enter saliva by diffusing through the cells of the salivary glands and that their concentration in saliva does not depend on the rate of saliva production. Conjugated steroids enter saliva via "ultrafiltration" through the tight junctions between the acinar cells, and their concentration in saliva is highly flow-rate dependent. Thyroxin and choriogonadotropin enter saliva via the ultrafiltration route or by contamination of the saliva by plasma or gingival fluid. We conclude that the salivary concentration on unconjugated steroids may usefully reflect the concentration of free (nonprotein-bound) steroids in plasma. Conversely, the concentration of conjugated steroids, thyroxin, and protein hormones such as choriogonadotropin in saliva probably does not reflect their concentration in plasma in any clinically useful way.

13)Salivary Steroid Assays for Assessing Variation in Endocrine Activity; Riad-Fahmy D, Read GF and Walker RF. J Steroid Biochem 19: 265-272, 1983

Salivary sampling regimens are non-invasive, and therefore facilitate dynamic tests of hormone function and assessment of biological rhythms. Concentrations of neutral steroids in saliva are independent of flow rate and appear to reflect the non-protein-bound, ‘free’ fraction. Comparison of replicate determinations of quality control pools with determinations of samples collected at 2 min intervals allows the significance of short-term fluctuations in cortisol and testosterone secretory activity to be estimated. Samples collected at 15 min intervals provide a convenient way to estimate circadian rhythms, particularly in young children. Determination of salivary progesterone concentrations in samples collected by women daily, over extended periods of time, provides a valuable means of assessing ovarian function. Such assays may be used to monitor ovulation-induction therapy.

14) Steroid assays in saliva: a method to detect plasmatic contaminations; G. Lac, N. Lac and A. Robert. Archives Internationales de Physiologie, de Biochimie et de Biophysique, 101, 257-262, 1993.

Plasmatic and salivary levels of six steroid hormones in adult males and females are given and compared to the data of the literature. These steroids are: cortisol (F), dehydroepiandrosterone (DHA) and its sulphate (DHAS), androstenedione (A), testosterone (T) and 11$OH androstenedione (OHA). The salivary assay of the last compound is an original.

The correlations between salivary and plasmatic values are presented and confirm that this method is a reliable alternative for hormonal investigations. From these data and from those of the literature, the salivary versus plasmatic ratio are calculated. From the fact that high concentrations of DHAS in saliva generally stem from blood contamination, we derive a method to estimate the amount of this contamination and its impact on other steroids measured on the same saliva sample.

15) Salivary Testosterone Measurements: Collecting, Storing, and Mailing Saliva Samples; Dabbs JM Jr. Physiology and Behavior 49: 815-817, 1991

Salivary testosterone measurements can be especially useful in field studies, but reliable ways of collecting and handling samples need to be established. Using cotton dental rolls to collect saliva leads to inflated testosterone scores. Sugar free gum can be used satisfactorily to stimulate saliva among both male and female subjects. Leaving unpreserved saliva samples at room temperature for 2 weeks or mailing them unrefrigerated is satisfactory for male subjects but leads to inflated scores for female subjects.

16) The Determination of Bio-available Testosterone; Wheeler MJ. Ann Clin Biochem 37: 50-55, 1997

17) Assessment of Testicular Function by the Radioimmunoassay of Testosterone in Saliva; Walker RF, Wilson DW, Read GF and Riad-Fahmy D. Int J Androl 3: 105-120, 1980

A sensitive RIA (500 fg/tube) has been developed for assay of testosterone in male saliva and extensively validated. In normal male saliva samples, morning concentrations (368+/-167 pmol/l) were significantly higher than evening samples (212+/-132 pmol/l). The circadian rhythm was confirmed by SOSINOR analysis. Levels of testosterone in saliva, in response to HCG stimulation, accurately reflected the increase observed in matched plasma samples. Synacthen administration, although increasing circulating cortisol levels, caused no significant change in plasma and salivary testosterone concentrations. Prostatic cancer patients on diethylstilboestrol therapy had low salivary (47-122 pmol/l) and plasma (1.0-2.8 nmol/l) testosterone concentrations.

Correct assessment of testicular function following stimulation and treatment regimens requires multiple sampling. Since saliva samples are easily collected by non-invasive techniques they represent an attractive alternative to plasma for evaluation of androgenicity.

18) Analysis of Testosterone and Dehydroepiandrosterone in Saliva by Gas Chromatography-Mass Spectrometry; Gaskell SJ, Pike AW, and Griffiths K. Steroids 36: 219-228, 1980

Testosterone and 3ß-hydroxyandrost-5-en-17-one (dehydroepiandrosterone) have been identified in human parotid fluid and saliva by gas chromatography-mass spectrometry/selected ion monitoring analyses of the t-butyldimethylsilyl ether and methyl oxime, t-butyldimethylsilyl ether derivatives. High specificity of analysis has been achieved by the use of high mass spectrometric resolution or by the monitoring of metastable peaks. Quantitative analysis indicate concentrations of both unconjugated testosterone and unconjugated dehydroepiandrosterone in the range 200-800 pmol/l in the saliva and parotid fluid of the normal males examined. These represent 1.5-7.5% of the concentrations of the steroids in blood plasma taken from the same subjects.

19) Bioavailable Testosterone and Depressed Mood in Older Men: The Rancho Bernardo Study; E Barrett-Connor, DG Von Muhlen and D Kritz-Silverstein. J Clin Endocrinol Metab 84: 573-577, 1999

A cross-sectional population-based study examined the association between endogenous sex hormones and depressed mood in community-swelling older men. Participants included 856 men, ages 50-89 yr, who attended a clinic visit between 1984-87. Total and bioavailable testosterone, total and bioavailable estradiol, and dihydrotestosterone levels were measured by radioimmunoassay in an endocrinology research laboratory. Depressed mood was assessed with the Beck Depression Inventory (BDI). Levels of bioavailable testosterone and bioavailable estradiol decreased with age, but total testosterone, dihydrotestosterone, and total estradiol did not. BDI scores increased with age. Low bioavailable testosterone levels and high BDI scores were associated with weight loss and lack of physical activity, but not with cigarette smoking or alcohol intake. By linear regression or quartile analysis the BDI score was significantly and inversely associated with bioavailable testosterone (bothP’s=0.007), independent of age, weight change, and physical acitivity; similar associations were seen for dihydrotestosterone (P=0.048 and P=0.09, respectively). Bioavailable testosterone levels were 17% lower for the 25 men with categorically defined depression than levels observed in all other men (P=0.01). Neither total nor bioavailable estradiol was associated with depressed mood. These results suggest that testosterone treatment might improve depressed mood in older men who have low levels of bioavailable testosterone. A clinical trial is necessary to test this hypothesis.

20) The Measurement of Hormones in Saliva: Possibilities and Pitfalls; Vining RF, McGinley RA. J Steroid Biochem 27: 81-94, 1987

The easy stress-free, non-invasive nature of saliva collection makes it one of the most accessible body fluids and it is potentially of value in studying normal human physiology as well as pathology. Measurements of salivary hormone levels will usually only be of value if they reflect the plasma level of the hormone and the relationship between the saliva and plasma levels of many hormones have been studied by a number of groups. The measurement of the salivary level is a valuable clinical tool for some hormones (e.g. cortisol, oestriol, progesterone), is of little value for others (e.g. cortisone, dehydroepiandrosterone sulphate, thyroxine, pituitary hormones) and for many others the saliva/plasma relationship is not yet sufficiently understood to assess the value of the salivary measurement. As well as reviewing the state of our knowledge of the salivary concentration of many hormones this review outlines a number of "rules of thumb" concerning the presence of hormones in saliva, their saliva/plasma relationship and the potential usefulness of assays of their salivary concentration.

These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease.



 

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