No Evidence for the Presence of 1,4-Androstadiene-3,17-dione in Rhodiola rosea

Reviewed: Da J, Yang L, Wu WY, Guo DA, Panossian AG. The pseudoendogenous anabolic steroid 1,4-androstadiene-3,17-dione does not occur naturally in Rhodiola rosea L. radix and rhizome. Phytochem Lett. 2018;23:155-163.

Keywords: Anabolic steroids, 1,4-androstadiene-3,17-dione, GC-MS, Rhodiola rosea, UHPLC-QTOF-MS

This research project was initiated after a paper by researchers from the German Sport University (Cologne, Germany) found the anabolic steroids 4-androstene-3,17-dione, 1,4-androstadiene-3,17-dione, and dehydroepiandrosterone (DHEA), generally between less than 10 ng/g – 312 ng/g although one product had 8.74 μg/g of DHEA, in 14 products labeled to contain Rhodiola rosea (syn. Sedum rosea, Crassulaceae).¹ Some of these products were advertised to improve performance, or for weight management. The authors hypothesized that these anabolic steroids might be from accidental contamination, or might occur naturally in the rhodiola roots.

In order to determine if rhodiola produces 1,4-androstadiene-3,17-dione, Da et al. analyzed authentic rhodiola roots and rhizomes from Canada, Norway, and Russia by ultrahigh-performance liquid chromatography with ultraviolet detection (UHPLC-UV), UHPLC coupled to quadrupole time-of-flight mass spectrometry (QTOF-MS), and gas chromatography (GC) with MS detection. Using the validated UHPLC-QTOF-MS method, 1,4-androstadiene-3,17-dione could be detected at concentrations equal to, or higher than 1.6 ng/g of test material. The detection limits for the UHPLC-UV and GC-MS methods were 978 ng/g and 2.44 ng/g, respectively. The anabolic steroid was not found in any of the rhodiola materials, suggesting that the rhodiola plant does not produce 1,4-androstadiene-3,17-dione, and that the results published by Walpurgis et al.¹ point to a problem in the manufacturing process of the products in question, or to deliberate adulteration with anabolic steroids in order to enhance the benefits of these products in consumers looking for a herbal supplement with performance-enhancing effects.

Comment: The task of providing evidence for the absence of a particular chemical in a plant species is not as easy as one might think. Since the phytochemical composition is influenced by many factors, such as minor variations in the genetic makeup producing the metabolites (i.e., the chemotype), location, and climatic aspects, it is important to evaluate a number of individual plant specimen from several locations. It is a matter of discussion if three different analytical methods are essential to verify the absence of a compound, or if the method providing the lowest limit of detection, if properly validated, would suffice. However, the use of two independent laboratories to verify the results has to be commended, representing a robust piece of evidence that 1,4-androstadiene-3,17-dione is not naturally found in Rhodiola rosea. However, this makes the data published by Walpurgis et al. even more concerning, since it points to either sloppy manufacturing or intentional adulteration of the rhodiola-containing products.

References

1. Walpurgis K. Schultze G, Mareck U, Geyer H, Schänzer W, Thevis M. Detection of endogenous and pseudoendogenous steroids in dietary supplements containing Rhodiola rosea. In: Schänzer W, Thevis M, Geyer H, Mareck U eds. Recent Advances in Doping Analysis, vol 24. Köln, Germany: Sportverlag Strauß; 2016:131–134.