Identification of S22 (ostarine) in human nails and hair using LC‐HRMS. Application to two authentic cases

Abstract

Ostarine, also known as S22 or MK2866 and enobosarm, is a selective androgen receptor modulator (SARM). It has high anabolic potency, in addition to limited androgenic effects. At this time, ostarine has no therapeutic use, but can be abused for performance-enhancing purposes using the oral route, at dosages of 10-25 mg per day. As the drug can easily be obtained via the Internet or some fitness centers, athletes and more and more amateurs can use it without undergoing the deleterious physiological side effects that are generally associated with testosterone-related compounds. Since 2008, the World Anti-Doping Agency (WADA) has prohibited SARMs at-all-times in the category of "other anabolic agents" under section S1.2 of the WADA List. In a case of trafficking/abuse, ostarine was identified in nail (subject 1) and hair (subject 2) by LC/HRMS after incubation of 50 mg of matrix in a pH 9.5 buffer, followed by extraction with organic solvents. The drug was quantitated by LC-MS/MS. Ostarine tested positive at 61 pg/mg (toenails) and 111 pg/mg (fingernails) for subject 1. Ostarine was present at 146, 168, 93, and 101 pg/mg in the 4 × 3 cm hair sections of subject 2, clearly demonstrating long-term use.

Peroxisome Proliferator-Activated Receptor Delta Agonist (PPAR- δ) and Selective Androgen Receptor Modulator (SARM) Abuse: Clinical, Analytical and Biological Data in a Case Involving a Poisonous Combination of GW1516 (Cardarine) and MK2866 (Ostarine)

“…The biological specimens (blood, urine and hair) were tested by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS), using previously described methods for ostarine [ 18 , 19 ] and cardarine [ 20 ]. Briefly, 1 mL of blood or hydrolyzed urine (with 20,000 UI of ß-glucuronidase at pH 5.2) was extracted in 1 mL borate pH 9.5 buffer and 5 mL of diethyl ether/dichloromethane/hexane (50/30/20, v / v ) in presence of 100 ng of bicalutamide-d 4 used as internal standard.…”

“…As a consequence, in the limited case reports involving SARMs or PPAR- δ, there is no toxicological data (identification of the substance and evaluation of its concentration). Testing for ostarine [ 9 , 18 , 19 , 20 ] and GW1516 [ 21 ] in human hair is the privilege of our laboratory as no other citation is available in the scientific literature.…”

Annual banned‐substance review: Analytical approaches in human sports drug testing 2019/2020

“…Conversely, M1a was found only when higher amounts were administered, and compared with all other target analytes, M1a was the most transient marker for ostarine intake, thus indicating a more recent exposure to the drug candidate. In contrast, the use of pharmacologically relevant amounts of ostarine was shown to be traceable in human hair and nails for months when utilizing an LC‐HRMS/MS‐based approach offering an LOD of 0.5 pg/mg of keratinous matrix 97 . Ostarine was chromatographed on a C‐18 column (150 × 2.1 mm, 1.8 μm particle size) with 0.1% formic acid (solvent A) and acetonitrile (containing 0.1% formic acid, solvent B), ionized by negative ESI, and measured in full scan mode with a collision energy ramp from 10 to 40 eV.…”

Detection of the selective androgen receptor modulator GSK2881078 and metabolites in urine and hair after single oral administration

“…Due to possible long‐term detection in hair, valuable methods are established to differentiate food contamination from clenbuterol abuse 10 and to identify administered intact compounds, for example, testosterone esters in connection to elevated T/E ratio in urine 11 . With regard to SARMs, only few studies investigated their incorporation in the hair matrix, among them LGD‐4033, SARMs S4 and S22 12–14 …”

“…11 With regard to SARMs, only few studies investigated their incorporation in the hair matrix, among them LGD-4033, SARMs S4 and S22. [12][13][14] Aim of the present study was the elucidation of the in vivo metabolism of GSK2881078 to identify metabolites as potential targets for doping analysis. After oral administration of GSK2881078 by one healthy volunteer, urine samples were collected up to 42 days post administration.…”