picture of testing flow chart

This is a strange study that brings up more questions than answers really. The levels found so low, they’re physiologically irrelevant which suggests due to cross incrimination vs intentional. Still, for athletes who get tested and such, that could be an issue. Note that 4-androstenedione (4AD) is still sold grey market and is a pro-hormone vs an AAS as most think of AAS. 4AD was introduced by chemist Patrick Arnold back in the day. That may explain minute amounts ending up in some whey products as cross contamination and why no methyl test found.

What brands did they test? It’s also interesting to note the study was done in Iran, so were these common protein supplements taken off the shelve sold in Iran? Is that something users in the US should be concerned about? I tend to doubt it’s an issue among the major manufactures sold in the US, but may be another good reason to stick to trusted brands who are more likley to test their products and either produce it themselves or use quality contract manufacturers to make their products, especially tested athletes.

Survey of protein-based sport supplements for illegally added anabolic steroids methyltestosterone and 4-androstenedione by UPLC-MS/MS

J.Steroids. 2020


•Simple and rapid method was used to extract methyltestosterone and 4-androstenedione from protein supplement matrices.

LC-MS/MS method with matrix matched calibration was validated for simultaneous quantification of the interested anabolic hormones.•

4-androstenedione was found in 36.67% of whey-based supplements; none of them had methytestosterone.


There is some evidence that marketable supplements contain hormones not declared on the product label. The presence of these androgenic anabolic steroids (AAS) in sports supplements can be considered an adulteration and affect the health of consumers, who are predominantly athletes.

This study aimed to measure anabolic hormones (methyltestosterone and 4-androstenedione) in sport supplements. Ultra Performance Liquid chromatography coupled mass spectrometry (UPLC-MS/MS) with electrospray ionization (ESI) in positive mode was employed under the Multiple Reaction Monitoring (MRM) ion program. To overcome matrix effects and quantify the selected analyte, the calibration curve was made using Matrix Match method. The LOQ and LOD were 1 ng/g and 0.3 ng/g for both analytes. The recovery of 4-androstenedione and methyltestosterone was in the range of 86.87-107.35 and 77.31-113.98, respectively.

In terms of reproducibility, CV % for 4-androstenedione and methyltestosterone ranged from 6.56-16.87% and 1.45-15.12%, respectively. 4-androstenedione was found in 11 samples including 9 whey as 1.578 ± 0.154 ng/g and 2 whey albumin samples with an amount of 1.134 ng/g and 1.474 ng/g.

Consequently, continuous controlling of sport supplements comprising intentionally or unintentionally added androgens could be important for health and discuss in the context of compliance with anti-doping.

  1. Lee 3 years ago

    I would think most weight lifters, bodybuilders and athletes wouldn’t mind a “little extra” in their whey. I sure wouldn’t mind a “boost” in my whey. But I’m 51 and need all the help I can get. Lol.

    Forgive me if I sound selfish. .

  2. rodney Burke 3 years ago

    um-m wouldn’t putting male steroids in protein powder be harmful to women? Especially those who are past menopause?-

    • Author
      Will Brink 3 years ago

      It would be potentially harmful to anyone using it if the doses are high enough. What was detected is not an anabolic steroid and at levels detected, a non issue. It may however, result in a pos test for tested athletes. It should not be found in protein powders regardless of course.

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