Can Sports Cars Increase Testosterone?

According to published reports by Hiscox UK in 2008,  just listening to the sound of a sports car increased salivary testosterone levels in male and female participants.

Another study, by Gad Saad et al, measured a testosterone increase in men who were given the opportunity to drive a $150,000 Porsche 911 Carrera Cabriolet for an hour. When participants drove a decade-old Toyota Corolla, a slight decline in testosterone levels was observed. According to the author, this implies conspicuous consumption can reflect social status and therefore influence hormone levels.

I found the idea interesting and amusing that driving a sports car (or even hearing one) can actually increase testosterone.  A recent report of testosterone levels in the general male U.S. population suggests that, contrary to conventional wisdom, testosterone levels do not decline precipitously after the age of 30. In fact, there appears to be a visible (though not statistically significant) increase in testosterone levels in men in their 50s and 60s.

These data, coupled with the high prevalence of testosterone replacement therapy in the U.S., make it clear that accurate and precise testosterone measurement is very much in need when establishing population-based reference intervals and recommendations for testosterone replacement therapy.

As an employee of Waters, I began to consider the various methods used to measure testosterone levels, as measuring an increase would require highly sensitive analytical methods.  It’s probably not surprising that measuring hormone levels in saliva and other matrices can be challenging. Traditionally, ligand binding assays (LBAs) have been employed to measure free testosterone.

Although LBAs have been around for years, they have struggled to measure trace testosterone levels in saliva, particularly in women and children who can have lower ranges between 35 and 50 ng/dL compared to 600 and 700 ng/dL in men.  Also, the variability and inaccuracy of LBAs often compromise analytical testing reliability, mainly because antibodies used in LBAs can pick up active and inactive forms of testosterone.

Advances in liquid chromatography and mass spectrometry (LC-MS) have led to highly sensitive research applications that have advantages over LBAs. LC-MS is highly sensitive, allows for multiplexing (multiple hormones), and does not confuse active and inactive molecules, which opens the gates for easier biomarker quantification.

According to a 2014 publication by BG Keevil et al: “Salivary testosterone (Sal-T) measurements have not gained general acceptance in the clinical and research communities… largely due to uncertainties over the accuracy and reliability of [LBA] Testosterone (T) assays… related to difficulties in obtaining antibodies for the development of suitably sensitive and specific T immunoassays…. Recent studies have described… LC-MS/MS analysis of Sal-T in males but the assays required either derivatization or large sample volumes to improve sensitivity…. To date, there have been no studies using LC-MS/MS to measure Sal-T in both males and females.”

BG Keevil et al reportedly published the first study using LC-MS to systematically validate Sal-T measurement in both male and female adults, using a Waters ACQUITY UPLC System and the Xevo TQ-S Mass Spectrometer  (LLOQ 5 pmol/L).

Other advantages of using LC-MS are the detection of multiple hormones in a sample and increased analytical sensitivity.

Innovations in microflow LC, such as with the ionKey/MS System, offer increases in analytical sensitivity and solvent savings.  For example, R.M. Buttler et al simultaneously measured salivary testosterone, and rostenedione and DHEA in the 0.10 nmol/L range using a Waters TQ-S.

By leveraging the new ionKey/MS,  they yielded a 100- to 400-fold increase in on-column analytical sensitivity while at the same time decreasing solvent usage by 150 fold, as compared to standard flow methods. This allowed for simplification of the steroid extraction procedure, which in turn streamlined the sample preparation and reduced per-sample cost.

Regardless of the factors that influence hormone levels, accurate quantification is essential in clinical research, and LC-MS is a highly sensitive and accurate approach to measurement. New innovations such as the ionKey/MS System and high performance mass spectrometry will continue to expand the possibilities for detection.

And who knows? Maybe testosterone replacement therapy can be supplemented by a recording of the engine sounds of a Lamborghini?

Read up on the ability to quantify steroid compounds in human serum using LC-MS:

• Application note:  Multiplexed Analysis of Steroid Hormones Using ionKey/MS
• Poster:  Development of a High-sensitivity Micro LC-MS Method for Estradiol Clinical Research

Learn more about Waters Analytical Research Solutions for Health Science Research by visiting our website at legacy-stage.waters.com/translation.