Analysis of Anabolic Agents in Whey Protein by Gas Chromatography Coupled to Triple Quadrupole Mass Spectrometry
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Analysis of Anabolic Agents in Whey Protein by Gas Chromatography Coupled to Triple Quadrupole Mass Spectrometry Bruno Ruiz Brandão da Costa 1 & Monica Costa Padilha 1 & Lucas Martins Lisandro Rodrigues 1 & Rafaela Rocha Roiffé 2 & Márcia Nogueira da Silva de la Cruz 1 Received: 17 January 2020 / Accepted: 28 June 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Whey protein-based supplements are one of the most consumed products to improve performance, and they are, generally, recognized as being safe. However, recent studies indicated different types of adulterations in these products, including the presence of diuretics and stimulants, allowing the possibility of contamination by other substances with pharmacological activities, such as anabolic agents. In this sense, the aim of this study was to develop and validate a method for the screening of 46 anabolic agents in whey protein. The analytical method consisted of sample preparation using liquid-liquid extraction, followed by analytes derivatization and, finally, analysis by gas chromatography coupled to triple quadrupole mass spectrometry in two different experiments: multiple reaction monitoring and full scan. After validation, the method proved to be specific, accurate, and precise; no carry over effect was observed, and the limit of detection values varied between 0.5 and 100 ng g−1. The analysis of 44 commercial samples did not detect the presence of any anabolic agent. However, full scan analysis detected caffeine in 14 samples and bisphenol A in one of them. Therefore, these results indicate that the analyzed samples are safe concerning the presence of the evaluated anabolic agents, but it does not mean that they are free from the presence of other substances with pharmacological activities which are not declared on label. Keywords Whey protein . Contamination . Quality control . Nutritional supplements . Anabolic agents . Doping
Introduction The use of nutritional supplements has been growing continuously over the last years (Barnes et al. 2016; Garthe 2019; Kantor et al. 2016). As example, by 2018, the worldwide market was evaluated in US$115 billion and is estimated at more than US$190 in 2025 (GVR 2019). Concerns for wellness, fitness, and enhanced quality of life; illness prevention, compensation for inadequate diet, and performance enhancement are the major factors to justify its consumption by athletes and non-athletes (Binns et al. 2018). In this sense, one of the most used products are whey protein (WP)-based supplements (Chen et al. 2014; Petróczi et al. 2007). WP is the soluble protein fraction in milk serum, which is obtained as a by-product during cheese production, specifically * Márcia Nogueira da Silva de la Cruz [email protected] 1
Instituto de Química, UFRJ, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 11941-598, Brazil
2
IME, Instituto Militar de Engenharia, Urca, Rio de Janeiro, RJ 22290-270, Brazil
during the casein precipitation step (Garrido et al. 2016; Haraguchi et al
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