INTRODUCTION We examined urinary and serum concentrations of formoterol in asthmatic and healthy individuals after a single dose of 18 μg inhaled formoterol and after repeated inhaled doses in healthy individuals. Results were evaluated using the World Anti-Doping Agency (WADA) 2012 threshold for formoterol. METHODS On the day of this open-label, crossover study, 10 asthmatic subjects who regularly used beta2-agonists and 10 healthy participants with no previous use of beta2-agonists received a single dose of 18 μg formoterol. Further, 10 nonasthmatic participants inhaled 18 μg formoterol every second hour until obtaining a total of 72 μg, which is twice the maximum daily dose (36 μg formoterol) permitted by the WADA. Blood samples were collected at baseline, 30 min, 1, 2, 3, 4, and 6 h after the first inhalation. Urine samples were collected at baseline, 0-4, 4-8, and 8-12 h after the first inhalation. RESULTS Median urine concentration, corrected for specific gravity, after the single-dose administration peaked during 0-4 h after inhalation at a maximum of 7.4 ng·mL(-1) in asthmatic subjects and 7.9 ng·mL(-1) in healthy subjects. Median urine concentration after repeated doses peaked during 4-8 h after inhalation of a total of 72 μg formoterol at a maximum of 16.8 ng·mL(-1) in healthy participants. The maximum individual concentration of 25.6 ng·mL(-1) was found after inhalation of a total of 72 μg formoterol. CONCLUSIONS We found no significant differences in urinary and serum concentrations of formoterol between asthmatic and healthy subjects. We found high interindividual variability in the concentrations in all groups. Our data support the WADA 2012 urinary threshold of 30 ng·mL(-1) formoterol as being an adverse analytical finding.

Changes in the urinary steroid profile may be an indication of the use of testosterone or its precursors. The concentrations and ratios of selected steroids show a large inter-individual variation, and the screening approach relying on population based criteria lacks the discriminating power to detect the administration of testosterone in low doses. Genetic variation is the most important cause of inter-individual variability in the excretion of testosterone metabolites. The enzyme UGT2B17 is important in the glucuronidation of testosterone. A deletion in the gene encoding for this enzyme causes a reduced excretion of testosterone glucuronide. The aim of this study was to investigate how the steroid profile is affected in subjects with different UGT2B17 genotypes after dermal application of testosterone gel and injection of testosterone enanthate. Ten male volunteers were included in the study. Urine samples were collected prior to the administration of testosterone and during the applications. In addition a blood sample was collected for the determination of their UGT2B17 genotype. The urinary steroid profiles were analysed by GC-MS. Selected samples were also analysed by isotope ratio mass spectrometry (IRMS). The results show that the T/E ratio was affected by administration of testosterone, but only to a limited extent for dermal preparations. Even in individuals with small changes in T/E ratio following testosterone administration, analysis by IRMS confirmed the presence of testosterone and metabolites of exogenous origin. Introduction Changes in urinary steroid profile may indicate the use of testosterone or its precursors. The steroid concentrations and selected ratios show a large inter-individual variation. Genetic polymorphism is the most important cause of inter-individual variability in the excretion of testosterone metabolites [1]. The enzyme UGT2B17 is important in the glucuronidation of testosterone. A deletion in the gene encoding for this enzyme results in reduced excretion of testosterone glucuronide [1]. The aim of this study was to investigate if the steroid profile is affected in subjects with different UGT2B17 genotypes after dermal application of testosterone gel and injection of testosterone enanthate in low doses. The disclosure of such administration was also investigated by isotope ratio mass spectrometry (IRMS). Experimental Ten male volunteers, aged 21-29 years, were included in the study. Prior to the administration of testosterone, five urine samples were collected from each participant, in order to establish their baseline steroid profiles. In addition, a blood sample was collected for the determination of their UGT2B17 genotype. Testosterone gel (50 mg/day) was applied once daily for seven consecutive days. Urine samples were collected daily during the application period and for the following seven days. After an eight week wash-out period, the participants received a single intramuscular injection of testosterone enanthate (125 mg). Urine samples were collected for 14 days. The urinary steroid profiles were analysed by GC-MS by the established steroid screening procedure modified by introducing five point calibration curves for the selected endogenous steroids. ,2

Objective: Data on pharmacokinetics of inhaled and oral salbutamol in elite athletes with asthma are needed to differentiate between therapeutic use and doping in doping control.Design: An interventional open-label crossover.Setting: Respiratory Research Unit, Copenhagen University Hospital, Bispebjerg.Participants: Eight elite athletes with asthma and 10 nonasthmatic subjects aged 18 to 33 years.Intervention: Administration of 0.8 mg of inhaled salbutamol and 8 mg of oral salbutamol separated by 14 days.Main Outcome Measures: Urine concentration of free salbutamol.Results: Maximum urine concentrations peaked in the period of 0 to 4 hours after the administration of inhaled and oral salbutamol in both groups. Median concentrations after inhaled salbutamol and oral salbutamol were 401.6 and 2108.1 ng/mL in healthy subjects and 334.9 and 2975.2 ng/mL in elite athletes with asthma. There were no significant statistical differences between the groups. One sample exceeded the World Anti-Doping Agency threshold value of 1000 ng/mL with a urinary salbutamol concentration of 1057 ng/mL 4 hours after inhalation, when no correction for urine specific gravity was done. When this sample was corrected for urine specific gravity, the result was 661 ng/mL.Conclusions: We found no significant difference in pharmacokinetic profile of inhaled and oral salbutamol between elite athletes with asthma and nonasthmatic subjects. Our results indicate that urine salbutamol concentrations should be corrected for urine specific gravity when evaluating doping cases.

Background: Salmeterol is a long acting β2agonist that is used in treatment of asthma. β2agonists are on WADA and IOC9s prohibited list, but salmeterol is allowed in therapeutic doses by inhalation. The prohibited list however contains no urinary limit for salmeterol, which gives athletes the opportunity to inhale unlimited doses of salmeterol. Large doses of β2agonists may have ergogenic effects and therefore a quantification of a urine salmeterol limit is necessary. Purpose: To find plasma and urinary concentrations of inhaled salmeterol by inhalation in therapeutic dose 100 μg in healthy and persons with asthma. To discuss a urinary concentration limit for inhaled salmeterol on the prohibited list. Methods: 10 persons with asthma (A) and 10 healthy subjects (C) were enrolled, age 24.6±3.9. The subjects underwent two visits. First visit was a pre-examination with a metacholine provocation and lung function test. On second visit the subjects inhaled 100 μg salmeterol (Serevent © ) as a single dose. Blood samples were acquired at baseline and 0.5, 1, 2, 3, 4 and 6 hours after administration. Urinary samples were collected at baseline and 4, 8 and 12 hours after administration. Plasma and urine samples were analyzed by liquid chromatography mass spectrometry. Results: The peak median urinary concentration was found after 4hrs reaching 0.38±0.26 ng mL -1 in A and 0.38±0.22 ng mL -1 in C. Peak median plasma concentration was 0.07±0.03 ng mL -1 for A and 0.06±0.03 ng mL -1 in C. No differences were found between the groups. Conclusions: Urine salmeterol peak 4h after administration by inhalation. We propose a salmeterol urine limit of 0.82 ng mL -1 in doping controls.

printing supported by . Visit Chiesi at Stand D.30 TUESDAY, SEPTEMBER 27TH 2011 residual asthmatic inflammation in the peripheral airway. The ECP levels in latephase sputum) 255.2±297.1ug/1 at study entry) significantly decrease 60.0±43.7 (p=0.038) and 50.7±48.4 (p0.049) at 4 and 8 weeks after switching to treatment with the FBC, respectively. The FeNo levels (76.0±69.4 ppb at study entry) also significantly decreased 29.1±15.7 (p=0.017) at 8 weeks. The R5-R20 and AX values of IOS parameters also significantly improved after 8 weeks. Conclusion: This study suggests that the FBC may give better control of residual eosinophilic inflammation in the distal airway compared to SFC therapy. P3952 Montelukast as add-on therapy may improve some indices of small airways involvement in uncontrolled asthmatics Laura Malagrinò1, Francesco Costa1, Lorenza Melosini1, Antonella Di Franco1, Giosuè Catapano2, Pierluigi Paggiaro1. 1Cardio-Thoracic and Vascular Department, University of Pisa, Pisa, Italy; 2Institute of Clinical Physiology,

Several peptide drugs are being manufactured illicitly, and in some cases they are being made available to the public before entering or completing clinical trials. At the request of Norwegian police and customs authorities, unknown pharmaceutical preparations suspected to contain peptide drugs are regularly subjected to analysis. In 2009, an unknown pharmaceutical preparation was submitted for analysis by liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS/MS). The preparation was found to contain a 29 amino acid peptide with a C-terminal amide function. Based on the interpretation of mass spectrometric data, an amino acid sequence was proposed. The sequence is consistent with a peptide currently marketed under the name CJC-1295. CJC-1295 is a releasing factor for growth hormone and is therefore considered a Prohibited Substance under Section S2 of the WADA Prohibited List. This substance has potential performance-enhancing effects, it is readily available, and there is reason to believe that it is being used within the bodybuilding community.

PURPOSEData on blood and urinary concentrations of salbutamol after inhalation and oral administration in healthy subjects are scarce. Accordingly, we examined the pharmacokinetics of inhaled and oral salbutamol in asthmatic subjects.METHODSWe enrolled 10 men aged 18-45 yr in an open-label study in which 0.8 mg of inhaled or 8 mg of systemic salbutamol was administered in a crossover design. All subjects had doctor-diagnosed asthma, used beta2 agonist when needed, and abstained from any medicine, beta2 agonist inclusive, for 14 d before visit. Urine was collected from all subjects (0-4, 4-8, and 8-12 h), and blood samples were taken at 0, 0.5, 1, 2, 3, 4, and 6 h after salbutamol administration.RESULTSMaximum urine concentration was reached during the first 4 h after administration of both inhaled and oral salbutamol. We found differences in median urinary concentrations (Cmax) of 260.9 and 2422.2 ng x mL(-1), respectively (P < 0.005). Urinary concentrations show high individual variability irrespective of the route of administration. Blood analyses showed a systemic exposure of salbutamol after both inhaled and oral salbutamol with peak concentration after inhalation before the oral intake (P < 0.05). A difference in median Cmax after inhalation and oral treatment was found: 1.75 and 18.77 ng x mL(-1), respectively (P < 0.05).CONCLUSIONSMedian urinary concentrations after oral administration of 8 mg of salbutamol were significantly higher than those after inhalation of 0.8 mg of salbutamol.