Saliva testing: Why it should be standard in the diagnosis of female hormone imbalance

For decades, female hormone levels were measured using blood tests.1 But blood sampling is invasive, expensive and is, at times, logistically difficult. Hence, there was a shift towards more convenient and inexpensive sample types like saliva.

Saliva-based tests are highly precise and painless. They are a reliable and proven method for evaluating female hormone levels. 2,3 This article elaborates on some of the challenges associated with saliva testing and its acceptance into the market.

Saliva testing: Why it should be standard in the diagnosis of female hormone imbalance

Image Credit: Tecan

Saliva-based diagnostics are currently employed to measure various physiological conditions, beyond sex hormone imbalances, like depression and stress, and sleep disorders.4-6 They are also used in sports medicine and occupational health.

Measuring hormone imbalance: The saliva testing workflow

As soon as the symptoms of a potential hormone imbalance have been identified, the medical practitioner would initially evaluate the level of specific hormones.

Normally blood samples are drawn, and serum-based tests are carried out. The search for less invasive and inexpensive techniques led to saliva-based testing.

Saliva testing initially showed variability in results owing to the lack of standardization of sample collection, transport and testing. However, SOPs are in place for everything from the collection devices to the laboratory tests.

The platforms for evaluating and analysing female hormone levels in saliva could be either mass spectrometry-based, for example, LC-MS/MS, or enzyme immunoassay (EIA, e.g., ELISA).

After the analysis of the samples and the comparison of the levels with normal ranges, results can be shared with the clinician, who later prescribes treatment like hormone replacement therapy (HRT).

Can saliva testing give the answers needed?

Saliva reveals the bioavailable, unbound hormone level (Figure 1). This might be only 2–5% of the total, however, it precisely represents the amount available to the body, in comparison to serum levels.7

When considering treatment options, it is the bioavailable level that is of the utmost importance. Using saliva ascertains that only active hormone is measured as saliva includes the free steroid fraction, whereas blood has total steroids.

Figure 1. Steroids in blood and saliva: the case for saliva testing. Image Credit: Tecan

Is saliva hormone testing easy to do?

Saliva is easy to self-collect using a standard device. Self-collection can be done as frequently as required to establish a hormonal profile — even if it is more than once daily.

When compared to the collection of blood samples in a clinic, this is much more convenient and takes up less time. The sample is later transported, stored and subjugated to rigorous, established SOPs in saliva testing laboratories.

Is saliva testing cost-efficient?

Saliva sampling is affordable compared to blood sampling, and the assays employed for hormone measurement — specifically EIA-based assays — are economically viable.

Saliva collection is 48% less expensive than blood collection when comparing costs that are unique to individual methods.8

So why is not everyone doing saliva hormone testing?

Compared to the health and wellbeing area, the overall market acceptance of saliva testing in the clinical community is lower and hence tests for female hormone imbalance are still serum-based.

However, complementary (or wellbeing) practitioners are pleased to perform saliva testing. However, they cannot prescribe, whereas medical practitioners who can prescribe, commonly choose to use blood.

A major reason for this is that a more holistic approach to health and wellbeing in the 21st century has seen an upsurge in complementary or alternative medicine.

Numerous online tests emerged with DNA and hormone tests leading the charge.9 Yet, a few of these tests lack a scientific foundation, let alone standardization.10

For saliva-based testing to be acknowledged in the clinic, the researchers with the empirical evidence that saliva-based testing works should share their knowledge and help a conservative audience who prefers to stay with the proven serum-based techniques to comprehend that saliva is a realistic and proven alternative.

Moreover, patients with internet access will first try to get an idea of their symptoms online, performing background research before consulting the clinician with internet-driven questions and expectations.

In such circumstances, if a medical practitioner has difficulty conveying the relative pros and cons of a certain technology that is not accepted in the clinic, they are more than likely to adhere to existing SOPs— and in this case, taking blood samples rather than saliva samples.

Saliva diagnostics has a long way to go before it becomes the “go-to” technology for certain traditional blood-based tests. However, the flexibility, user-friendliness and an increasing volume of scientific literature are all set to trigger this acceptance.

In an uncertain world of increasing healthcare costs that expects the development of holistic and personalized treatments, the market growth of a simple and inexpensive technique such as saliva testing is inevitable.


  1. Sufi, S. B., Donaldson, A., Gandy, S. C., Jeffcoate, S. L., Chearskul, S., Goh, H., Hazra, D., Romero, C., & Wang, H. Z. (1985). Multicenter evaluation of assays for estradiol and progesterone in saliva. Clinical chemistry, 31(1), 101–103. PubMed ID: DOI:
  2. Landman, A. D., Sanford, L. M., Howland, B. E., Dawes, C., & Pritchard, E. T. (1976). Testosterone in human saliva. Experientia, 32(7), 940–941. PubMed ID: DOI:
  3. Pujari, M., Bahirwani, S., Balaji, P., Kaul, R., Shah, B., Daryani, D., & Iqbal, S. (2012). Oral fluid nanosensor test: saliva as a diagnostic tool for oral health. Journal of the California Dental Association, 40(9), 733–736. PubMed ID :
  4. Bellagambi, F.G., Lomonaco, T., Salvo, P., Vivaldi F., Hangouet, M., Ghimenti, S., Biagini, D., Di Francesco, F. & Fuoco, R. & Errachid, A. (2020). Saliva sampling: Methods and devices. An overview. Trends in Analytical Chemistry. Elsevier. Volume 124, March 2020, 115781 Elsevier ID: DOI:
  5. Rahman, S. A., Kayumov, L., Tchmoutina, E. A., & Shapiro, C. M. (2009). Clinical efficacy of dim light melatonin onset testing in diagnosing delayed sleep phase syndrome. Sleep medicine, 10(5), 549–555. PubMed ID: DOI:
  6. Chojnowska, S., Ptaszyńska-Sarosiek, I., Kępka, A., Knaś, M., & Waszkiewicz, N. (2021). Salivary Biomarkers of Stress, Anxiety and Depression. Journal of clinical medicine, 10(3), 517. PubMed ID: DOI:
  7. Lewis J. G. (2006). Steroid analysis in saliva: an overview. The Clinical biochemist. Reviews, 27(3), 139–146. PubMed ID:
  8. DNA from saliva vs. blood – who wins the cost battle? Accessed 29 September 2021.
  10. Dr. Gonshor, personal observations.

About Tecan

Tecan is a leading global provider of automated laboratory instruments and solutions. Their systems and components help people working in clinical diagnostics, basic and translational research and drug discovery bring their science to life.

In particular, they develop, produce, market and support automated workflow solutions that empower laboratories to achieve more. Their Cavro branded instrument components are chosen by leading instrumentation suppliers across multiple disciplines.

They work side by side with a range of clients, including diagnostic laboratories, pharmaceutical and biotechnology companies and university research centers. Their expertise extends to developing and manufacturing OEM instruments and components, marketed by their partner companies. Whatever the project – large or small, simple or complex – helping their clients to achieve their goals comes first.

They hold a leading position in all the sectors they work in and have changed the way things are done in research and development labs around the world. In diagnostics, for instance, they have raised the bar when it comes to the reproducibility and throughput of testing.

In under four decades Tecan has grown from a Swiss family business to a brand that is well established on the global stage of life sciences. From pioneering days on a farm to the leading role our business assumes today – empowering research, diagnostics and many applied markets around the world

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Last updated: Feb 16, 2022 at 6:16 AM


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