The Reigning Underdog
Saliva also has its problems. Unfortunately, hormone saliva tests are not covered by insurance companies. So patients have to pay out-of-pocket, approximately $50 per test. Saliva hormone testing is also a new technology. It's been used only in the last 20 years and, therefore, not widely accepted by the medical community. Saliva testing also is not readily available in many laboratories. Furthermore, there's room for human error when gathering the saliva sample. Food or blood can easily contaminate the specimen.

The good news is that saliva collection is noninvasive, painless, relatively inexpensive and convenient for the patient. Saliva hormone testing, in fact, may increase accuracy by directly measuring biologically active or free hormones, which can be gathered by the patient without the assistance of a professional. Patients should take five samples of their saliva throughout the day to obtain an average of free hormones in the body.

When comparing saliva and serum methods, published studies have shown that a saliva sample is much more accurate than a serum sample.⁷ It is a reliable marker of testosterone bioavailability, principally for very low levels.⁸ (Serum tests are often unreliable in this area.) Saliva testing also can measure hormones such as testosterone in women or children and estrogen in men.⁹

In one study, 50 men, 55 women and 11 children were measured for serum testosterone by immunoassays and isotope-dilution gas chromatography spectrometry. None of the serum samples from the women and children were sufficiently reliable, in whom low (less than 1.7 nmol/L) and very low (0.17nmol/L) levels of testosterone are common.¹⁰ With proper education about the suitable methods of collecting and storing samples, we can ensure the accuracy and reliability of saliva testing. Here are seven common mistakes to avoid and improve accuracy.

Mistake #1. Overlooking factors that influence collected samples
Be aware of significant, natural ebbs and flows of hormone production, such as during the menstrual cycle. For example, high estrogen levels occur during ovulation, and high progesterone levels circulate through the body around day 21 of the menstrual cycle. When measuring hormones, we should always take these factors into consideration.

Mistake #2. Not obtaining all necessary samples
Hormones are produced in high levels of pulsatility and squirts on regular intervals, usually more in the morning and dropping throughout the day. Our clinical experience shows that five test samples give us an average of about 80% accuracy, while single sample collections at random reflects only a 20% or lower rate of accuracy. The first sample should be collected in the morning 1 hour after waking, with the remaining four samples collected every 3 hours after the last sample is collected. For hormone testing accuracy, all five samples must be gathered 3 hours apart.

Mistake #3. Using incorrect sample gathering utensils
Cotton swabs should never be used when collecting a sample. Cotton salivettes, which are balls of cotton with cotton strings attached, result in lower cortisol values. Cotton probably contains substances that interact with cortisol and bind hormones. This makes the cortisol unavailable for measuring. Spitting directly into the container using a small straw is preferred.

Mistake #4. Outside contamination in saliva sample
Before collecting saliva samples, patients must remember to wash their hands thoroughly with soap and water and use disposable gloves. Possible contamination will affect the accuracy of the test.

Mistake #5. Food contamination in saliva sample
Food contamination in the sample may cause inaccuracy. To avoid this, patients should rinse their mouths with clear water right after eating or drinking, and should not eat or drink 1 hour before a saliva sample collection. Rinsing the mouth with clear water 15 minutes before a saliva sample is acceptable.

Mistake #6. Blood contamination in saliva sample
Blood in the saliva sample, from bleeding gums or mouth sores, can contaminate the sample. The rejection point of contamination is when blood leaves a red tinge in the saliva specimen. To avoid blood in the saliva, patients should not brush or floss their teeth before collecting a saliva sample.

Mistake #7. Improper sample collection containers
A saliva sample can be soiled by improper collection containers, the most common of all contamination mistakes. Because saliva hormones are small nonpolar molecules and have a static electric charge, they tend to stick to plastic materials that have the opposite electric charge, such as polyethylene. The best plastic materials seems to be ultra-pure polypropylene and polystyrene, which do not bind the saliva hormone.

Polyethylene tubes and stoppers highly absorb free hormones, especially progesterone. They can absorb up to 87% of progesterone on the walls of the container. Glass seems to be best because it's completely absorption free. But it's not always practical because it can break during transport.

When comparing saliva and serum testing methods, we see that both have pros and cons. Saliva testing is easy and convenient. Multiple samples are gathered, which improves the chances of accuracy. Furthermore, saliva testing directly measures free hormone levels. It's also more accurate when measuring very low levels of free hormones.¹¹ Accurately measuring free hormone levels is essential for diagnosing, planning and maintaining proper treatment, as well as balancing hormones for total health.

Paul Ling Tai is a trained podiatric medical physician and board-certified surgeon with expertise in herbal compound engineering, research and development. He holds nine patents. He has served under two Michigan state governors and was recognized with a vice chairman's position on the Michigan Board of Podiatric Medical Licensing and as chief examiner for new podiatric physicians. He is a professor of integrative medicine at NYCPM, New York, a member of the International Hormone Society and the author of Cordyceps Miracles and 8 Powerful Secrets to Anti-Aging.

Disclosure: Tai indicates that he is CEO of Health Secrets USA, which provides salivary hormone testing.