Enzyme immunosorbent and mass spectrometric methods for the determination of testosterone and cortisol: comparative analysis

ˑ: 

PhD M.A. Dikunets1
G.A. Dudko1
Dr. Sc.Chem. E.D. Virus1
1Federal Science Center of Physical Culture and Sport (VNIIFK), Moscow

Objective of the study was to conduct a comparative analysis of enzyme-linked immunosorbent assay (ELISA) and mass spectrometric methods for determining cortisol and testosterone in human serum.
Methods and structure of the study. The samples were analyzed on a semi-automatic photometer BTS-350 (BioSystems, Spain) using ELISA kits and an ultra-fast liquid chromatography-mass spectrometer with a triple quadrupole LCMS-8060 (Shimadzu, Japan).
Results and conclusions. A high two-way correlation was revealed between the results of quantitative determination of cortisol and testosterone in serum using the compared methods. On average, cortisol concentrations measured photometrically using solid-phase ELISA were four times higher than those measured by mass spectrometry. To ensure the accuracy of the quantitative determination of steroid hormones in serum, it is necessary to use a chromatography-mass spectrometric method or first conduct a comparative analysis of the method used with a chromatography-mass spectrometric method to determine the conversion factor.

Keywords: cortisol, testosterone, enzyme immunoassay, gas chromatography-mass spectrometry.

References

  1. Bjerner J., Bormer O.P., Nustad K. The war on heterophilic antibody interference. Clin. Chem. 2005. 51(1). pp. 9-11.
  2. Horwath O., Apro W., Moberg M., et al. Fiber type-specific hypertrophy and increased capillarization in skeletal muscle following testosterone administration in young women. J. Appl. Physiol. 2020. 128(5). pp. 1240-1250.
  3. Instructions for Use. Cortisol ELISA. Available at: https://www.drg-diagnostics.de/files/eia-1887_ifu--cortisol_2019-05-02_e... (date of access: 02.17.2023).
  4. Ismail A.A.A. A radical approach is needed to eliminate interference from endogenous antibodies in immunoassays. Clin. Chem. 2005. 51(1). pp. 25-26.
  5. Keevil B.G. LC-MS/MS analysis of steroids in the clinical laboratory. Clin. Biochem. 2016. 49(13-14). pp. 989-997.
  6. Ketha S.S., Singh R.J., Ketha H. Role of mass spectrometry in clinical endocrinology. Endocrinol. Metab. Clin. North. Am. 2017. 46(3). pp. 593-613.
  7. Kraemer W.J., Ratamess N.A., Nindl B.C. Recovery responses of testosterone, growth hormone, and IGF-1 after resistance exercise. J. Appl. Physiol. (1985). 2017. 122(3). pp. 549-558.
  8. Kraemer W.J., Ratamess N.A. Hormonal responses and adaptations to resistance exercise and training. Sports Med. 2005. 35(4). pp. 339-361.
  9. Rämson R., Jürimäe J., Jürimäe T., Mäestu J. Behavior of testosterone and cortisol during an intensity-controlled high-volume training period measured by a training task-specific test in men rowers. J. Strength Cond. Res. 2009. 23(2). pp. 645-651.
  10. Viru A., Viru M. Cortisol-essential adaptation hormone in exercise. Int. J. Sports Med. 2004. 25(6). pp. 461-464.