Prevalence of synthetic cannabinoids in U.S. athletes: initial findings.

A number of synthetic cannabinoids such as JWH-018 and JWH-073 have been incorporated into "spice" products. Despite having labels warning against human consumption, the products are smoked for their cannabinoid-like effects and the extent of their use by athletes has not been adequately described. Urine samples collected from 5,956 athletes were analyzed by high-performance liquid chromatography-tandem mass spectrometry for the presence of JWH-018, JWH-073, and their metabolites. Metabolites of JWH-018 and/or JWH-073 were detected in 4.5% of the samples. Metabolites of JWH-018 and JWH-073, only JWH-018, and only JWH-073 were detected in 50%, 49%, and approximately 1% of positive samples, respectively. In total, JWH-018 metabolites were detected in 99% (50% + 49%) and JWH-073 metabolites were detected in approximately 50% (49% + 1%) of the positive samples. Parent JWH-018, JWH-018-2-OH-indole, and JWH-018-4-OH-indole were not detected in any of the samples. All samples in which JWH-073 metabolites were detected contained JWH-073-N-butanoic acid. Parent JWH-073 and its N-(4-OH-butyl), 4-OH-indole, 5-OH-indole, and 7-OH-indole metabolites were not detected. Given the number of synthetic cannabinoids that have been synthesized, their limited regulation, and the prevalence of JWH-018 and JWH-073 metabolites detected in the athletes, these compounds should remain a priority for anti-doping programs.

Screening indicators of dehydroepiandosterone, androstenedione, and dihydrotestosterone use: a literature review.

Because of their perceived and reported effects on self-image, muscle development, performance, and similar factors, anabolic-androgenic steroids (AAS) and their precursors are among the most abused substances by professional, amateur, and recreational athletes. However, AAS abuse is not limited to athletes, but is also prevalent in the workplace, especially those professions in which image, strength, and endurance are coveted attributes. The detection of many steroids in biological specimens is analogous to the detection of an abused drug such as cocaine. Identification of the parent drug or its characteristic metabolite(s) in a donor's sample with a drug screening technique and confirmation of the drug/metabolite with a suitable alternative technology provides evidence of use. These analyses and subsequent interpretive scenarios become far more complex when the ingested AAS is an endogenous compound such as dehydroepiandrosterone (DHEA), androstenedione (Adione), or dihydrotestosterone (DHT). These compounds and their metabolites are present in specimens such as urine as a course of our natural endocrine function. Therefore, it becomes much more challenging for the laboratory to establish testing and interpretative paradigms that can distinguish "normal" urinary profiles of these steroids and their metabolites from profiles indicative of exogenous use. Distinguishing "normal" from "abnormal" urine profiles is particularly challenging during screening when literally tens of steroids and their metabolites may be tested simultaneously in a single chromatographic analysis. The purpose of this paper is to review the relevant literature about DHEA, Adione, and DHT administration, detection, and interpretation specifically as it relates to changes in the urinary AAS profile that may be identified during the routine laboratory screening of donor urine specimens.

Normalization of urinary drug concentrations with specific gravity and creatinine.

Excessive fluid intake can substantially dilute urinary drug concentrations and result in false-negative reports for drug users. Methods for correction ("normalization") of drug/metabolite concentrations in urine have been utilized by anti-doping laboratories, pain monitoring programs, and in environmental monitoring programs to compensate for excessive hydration, but such procedures have not been used routinely in workplace, legal, and treatment settings. We evaluated two drug normalization procedures based on specific gravity and creatinine. These corrections were applied to urine specimens collected from three distinct groups (pain patients, heroin users, and marijuana/ cocaine users). Each group was unique in characteristics, study design, and dosing conditions. The results of the two normalization procedures were highly correlated (r=0.94; range, 0.78-0.99). Increases in percent positives by specific gravity and creatinine normalization were small (0.3% and -1.0%, respectively) for heroin users (normally hydrated subjects), modest (4.2-9.8%) for pain patients (unknown hydration state), and substantial (2- to 38-fold increases) for marijuana/cocaine users (excessively hydrated subjects). Despite some limitations, these normalization procedures provide alternative means of dealing with highly dilute, dilute, and concentrated urine specimens. Drug/metabolite concentration normalization by these procedures is recommended for urine testing programs, especially as a means of coping with dilute specimens.

Detection of bumetanide in an over-the-counter dietary supplement.

Bumetanide is a loop diuretic used clinically to treat heart failure, acute renal failure, high blood pressure, and edema. However, diuretics may also be used by athletes as masking agents and to decrease weight. Taken as masking agents, diuretics increase urine production and decrease urinary concentrations of banned performance-enhancing agents, such as anabolic steroids. StarCaps is an over-the-counter dietary supplement marketed as a diet aid. The manufacturer claims that the product contains only natural cleansing agents and emphasizes that it is free from traditional appetite suppressants such as sympathomimetic amines. However, no such disclaimer is made concerning diuretic agents. A single StarCaps capsule was administered to two male and two female volunteers, and their urine specimens were collected at discrete intervals (2, 4, 8, and 12 h) post administration. The specimens were analyzed by a high-performance liquid chromatography-mass spectrometry quadrupole (HPLC-MS) method, and bumetanide was detected in all specimens (4.6 to 351.3 ng/mL). Adjusting the bumetanide concentrations for creatinine content did little to normalize the excretion profiles. Bumetanide was also detected in the StarCaps capsules at concentrations approaching therapeutic doses. HPLC-quadrupole-time-of-flight mass spectrometry was used to confirm the presence of bumetanide in the urine samples and StarCaps capsules. The results showed that unregulated dietary supplements may put consumers at risk for unwitting consumption of prescription medications, and that it is possible for athletes to inadvertently test positive for bumetanide and face disciplinary actions.

Influence of phenobarbital on morphine metabolism and disposition: LC-MS/MS determination of morphine (M) and morphine-3-glucuronide (M3G) in Wistar-Kyoto rat serum, bile, and urine.

A simple LC-MS/MS method has been developed and validated for the simultaneous determination of morphine (M) and morphine-3-glucuronide (M3G) in rat serum, bile, and urine. Deuterated D3-M and D3-M3G were used as internal standards (IS) for M and M3G, respectively. Serum samples were processed by acetonitrile precipitation. Bile samples were prepared by solid-phase extraction (SPE) using Oasis MCX cartridges. Urine samples were directly analyzed after dilution with mobile phase. Chromatography was performed using a Luna C18 column (5 microm, 150x2.1 mm I.D.). The mobile phase consisted of acetonitrile (ACN) and 7.5 mM ammonium formate (pH 9.3) delivered from separate pumps with a simple gradient. The method was validated to quantify M in the range of 1-1000 ng/ml in bile and serum, and 0.025-25 microg/ml in urine. M3G was quantified in the range of 1-1000 ng/ml in serum, 0.1-100 microg/ml in bile, and 0.05-25 microg/ml in urine. The method was applied to study the pharmacokinetics and disposition of M and M3G in Wistar-Kyoto (WKY) rats, and the effect of phenobarbital (PB) on M and M3G disposition. M is metabolized to M3G at a lower rate in male than female rats leading to higher M levels and lower M3G levels in serum, urine, and bile of male than female rats. PB administration induces M glucuronidation to M3G in male, but not female WKY rats, and abolishes the gender differences in M and M3G pharmacokinetics.