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Cox H.D.,Sports Medicine Research and Testing Laboratory | Lopes F.,King's College London | Woldemariam G.A.,University of California at Los Angeles | Becker J.O.,University of Washington | And 7 more authors.
Clinical Chemistry | Year: 2014

BACKGROUND: Insulin-like growth factor 1 (IGF-1)7 is a key mediator of growth hormone (GH) action and a well-characterized biomarker of GH abuse. Current immunoassays for IGF-1 suffer from poor concordance between platforms, which makes comparison of results between laboratories difficult. Although previous work has demonstrated good interlaboratory imprecision of LC-MS/MS methods when plasma is supplemented with purified proteins, the interlaboratory imprecision of an endogenous protein in the nanogram-per- milliliter concentration range has not been reported. METHODS: We deployed an LC-MS/MS method to quantify serum IGF-1 in 5 laboratories using 5 different instruments and analyzed 130 healthy human samples and 22 samples from patients with acromegaly. We determined measurement imprecision (CV) for differences due to instrumentation, calibration curve construction, method of calibration, and reference material. RESULTS: Instrument-dependent variation, exclusive of digestion, across 5 different instrument platforms was determined to be 5.6%. Interlaboratory variation was strongly dependent on calibration. Calibration materials from a single laboratory resulted in less variation than materials made in individual laboratories (CV 5.2% vs 12.8%, respectively). The mean imprecision for 152 samples between the 5 laboratories was 16.0% when a calibration curve was made in each laboratory and 11.1% when a single-point calibration approach was used. CONCLUSIONS: The interlaboratory imprecision of serum IGF-1 concentrations is acceptable for use of the assay in antidoping laboratories and in standardizing results across clinical laboratories. The primary source of variability is not derived from the sample preparation but from the method of calibration. © 2013 American Association for Clinical Chemistry.


PubMed | German Sport University Cologne, Uppsala University and Sports Medicine Research and Testing Laboratory
Type: | Journal: Journal of pharmaceutical and biomedical analysis | Year: 2016

FG-4592 is a hypoxia-inducible factor (HIF) stabilizer, which can increase the number of red blood cells in the body. It has not been approved by regulatory authorities, but is available for purchase on the Internet. Due to its ability to improve the oxygen transportation mechanism in the body, FG-4592 is of interest for doping control laboratories, but prior to this study, little information about its metabolism was available. In this study, the metabolism of FG-4592 was investigated in a human doping control sample and in five in vitro models: human hepatocytes and liver microsomes, equine liver microsomes and S9 fraction and the fungus Cunninghamella elegans. By using liquid chromatography coupled to a Q-TOF mass spectrometer operated in MS


Hurley J.M.,University of Utah | Hurley J.M.,Sports Medicine Research and Testing Laboratory | West J.B.,University of Utah | West J.B.,Texas AgriLife Research Center | Ehleringer J.R.,University of Utah
International Journal of Drug Policy | Year: 2010

Background: Although cannabis is the most readily available and widely used illicit drug in the United States, there remains significant uncertainty about the importance of different production regions and trafficking patterns. Methods: We analysed 628 " retail" cannabis seizures from over 50 municipalities across the United States for hydrogen and carbon isotope ratios to predict their growth locations and environments. Results: Results are presented for 22 consolidated retail locations across the United States. Evaluation of specimens from within these retail areas suggested that cannabis seizures had region-dependent origins, often from both domestic and foreign sources, and although indoor growth was common in many areas, there was also regional dependence in the proportions cultivated under indoor versus outdoor conditions. Conclusion: Street-available cannabis exhibits region-specific trafficking patterns, both Mexican- and Canadian-grown cannabis are apparently widely available, and indoor-grown cannabis appears to be cultivated and trafficked in both warm and cool weather localities throughout the United States. © 2009 Elsevier B.V.


PubMed | Sports Medicine Research and Testing Laboratory and University of Utah
Type: Journal Article | Journal: PM & R : the journal of injury, function, and rehabilitation | Year: 2016

Historical reports of doping in sports date as far back as the ancient Greek Olympic Games. The anti-doping community considers doping in sports to be cheating and a violation of the spirit of sport. During the past century, there has been an increasing awareness of the extent of doping in sports and the health risks of doping. In response, the anti-doping movement has endeavored to educate athletes and others about the health risks of doping and promote a level playing field. Doping control is now undertaken in most countries around the world and at most elite sports competitions. As athletes have found new ways to dope, however, the anti-doping community has endeavored to strengthen its educational and deterrence efforts. It is incumbent upon sports medicine professionals to understand the health risks of doping and all doping control processes.


PubMed | University of Utah and Sports Medicine Research and Testing Laboratory
Type: Journal Article | Journal: Drug testing and analysis | Year: 2016

The laboratory profile of intranasal testosterone gel has not been previously reported from an anti-doping perspective. Because intranasal testosterone gel is newly available as a commercial product, we sought to examine the laboratory parameters following administration of this formulation, with particular attention to anti-doping guidelines. Five healthy and active male subjects were administered testosterone intranasal gel three times daily for four weeks, using a pattern of five consecutive days on, two days off. Urine was collected after each five-day round of drug administration and analyzed using a full steroid screen and isotope ratio mass spectrometry (IRMS). Windows of detection for elevated testosterone/epitestosterone (T/E) and other steroid ratios, World Anti-Doping Agency (WADA) athlete biological passport (ABP) findings, and IRMS results were analyzed in this study. In the 0-24h window post-administration, 70% of samples were flagged with a suspicious steroid profile and 85% were flagged as atypical passport findings according to the WADA ABP steroid module. In the 24-48h window, 0% of samples displayed suspicious steroid profiles while 40% resulted in atypical passport findings. IRMS testing confirmed the presence of exogenous testosterone in 90% and 40% of samples in the 0-24h and 24-48h windows post-administration, respectively. Additionally, IRMS data were analyzed to determine commonalities in the population changes in


Van Wagoner R.M.,Sports Medicine Research and Testing Laboratory | Satake M.,University of Tokyo | Wright J.L.C.,University of North Carolina at Wilmington
Natural Product Reports | Year: 2014

Covering: up to 2013 Dinoflagellates produce unique polyketides characterized by their size and complexity. The biosynthesis of a limited number of such metabolites has been reported, with studies largely hampered by the low yield of compounds and the severe scrambling of label in the isotopically-labeled precursors. Nonetheless, of the successful biosynthetic experiments that have been reported, many surprising and unique processes have been discovered. This knowledge has been accessed through a series of biochemical labeling studies, and while limited molecular genetic data has been amassed, it is still in the early stages of development. In an attempt to meet this challenge, this review has compared some of the biosynthetic processes with similar ones identified in other microbes such as bacteria and myxobacteria, with the idea that similar genes and enzymes are employed by dinoflagellates. This journal is © the Partner Organisations 2014.


Kelly B.N.,Sports Medicine Research and Testing Laboratory | Madsen M.,Sports Medicine Research and Testing Laboratory | Sharpe K.,University of Melbourne | Nair V.,Sports Medicine Research and Testing Laboratory | Eichner D.,Sports Medicine Research and Testing Laboratory
Drug Testing and Analysis | Year: 2013

Glycerol is an endogenous substance that is on the World Anti-Doping Agency's list of prohibited threshold substances due to its potential use as a plasma volume expansion agent. The WADA has set the threshold for urine glycerol, including measurement uncertainty, at 1.3 mg/mL. Glycerol in circulation largely comes from metabolism of triglycerides in order to meet energy requirements and when the renal threshold is eclipsed, glycerol is excreted into urine. In part due to ethnic differences in postprandial triglyceride concentrations, we investigated urine glycerol concentrations in a population of elite athletes competing in North America and compared the results to those of athletes competing in Europe. 959 urine samples from elite athletes competing in North America collected for anti-doping purposes were analyzed for urine glycerol concentrations by a gas chromatography mass-spectrometry method. Samples were divided into groups according to: Timing (in- or out-of-competition), Class (strength, game, or endurance sports) and Gender. 333 (34.7%) samples had undetectable amounts of glycerol (<1 μg/mL). 861 (89.8%) of the samples had glycerol concentrations ≤20 μg/mL. The highest glycerol concentration observed was 652 μg/mL. Analysis of the data finds the effects of each category to be statistically significant. The largest estimate of the 99.9th percentile, from the in-competition, female, strength athlete samples, was 1813 μg/mL with a 95% confidence range from 774 to 4251 μg/mL. This suggests a conservative threshold of 4.3 mg/mL, which would result in a reasonable detection window for urine samples collected in-competition for all genders and sport classes. © 2013 John Wiley & Sons, Ltd. Glycerol is a naturally occurring, prohibited substance potentially used by athletes in an effort to mask blood manipulation. Due to its endogenous nature, it is necessary to determine the threshold above which no athlete should naturally have a glycerol concentration in their urine. Data from a population (n=959) study of urine glycerol concentrations present in anti-doping control samples collected from elite athletes competing in North America in various disciplines as well as competition status are presented in this manuscript. © 2013 John Wiley & Sons, Ltd.


Cox H.D.,Sports Medicine Research and Testing Laboratory | Rampton J.,Sports Medicine Research and Testing Laboratory | Eichner D.,Sports Medicine Research and Testing Laboratory
Analytical and Bioanalytical Chemistry | Year: 2013

There is significant evidence that athletes are using recombinant human growth hormone (rhGH) to enhance performance, and its use is banned by the World Anti-Doping Agency and professional sports leagues. Insulin-like growth factor-1 (IGF-1) is the primary mediator of growth hormone action and is used as a biomarker for the detection of rhGH abuse. The current biomarker-based method requires collection and expedited shipment of venous blood which is costly and may decrease the number of tests performed. Measurement of GH biomarkers in dried blood spots (DBS) would considerably simplify sample collection and shipping methods to allow testing of a greater number of samples regardless of location. A method was developed to quantify intact IGF-1 protein in DBS by liquid chromatography-tandem mass spectrometry. A step-wise acid-acetonitrile extraction was optimized to achieve a sensitive assay with a lower limit of quantification of 50 ng/mL. IGF-1 remained stable at room temperature for up to 8 days, which would allow shipment of DBS cards at ambient temperature. In a comparison between plasma concentrations of IGF-1 and concentrations measured from venous and finger prick DBS, there was good correlation and agreement, r 2 of 0.8551 and accuracy of 86-113 % for venous DBS and r 2 of 0.9586 and accuracy of 89-122 % for finger prick DBS. The method is intended for use as a rapid screening method for IGF-1 to be used in the biomarker method of rhGH abuse detection. © 2012 Springer-Verlag Berlin Heidelberg.


PubMed | Sports Medicine Research and Testing Laboratory
Type: | Journal: Drug testing and analysis | Year: 2016

A new peptide, body protecting compound (BPC), BPC 157, and a variant of mechano-growth factor (MGF), MGF R23H, were identified in confiscated vials. BPC 157 has the amino acid sequence, GEPPPGKPADDAGLV, and is currently under investigation for the promotion of healing and recovery in a variety of tissues. In vitro metabolism experiments in plasma demonstrate that MGF R23H has good stability and should be detectable in urine, while BPC 157 forms a stable metabolite that should be detectable in urine. A weak cation exchange solid phase extraction method was validated for detection of BPC 157 in urine. The method has a limit of detection of 0.1ng/ml, precision of less than 20%, and good linearity, r


Cox H.D.,Sports Medicine Research and Testing Laboratory | Eichner D.,Sports Medicine Research and Testing Laboratory
Rapid Communications in Mass Spectrometry | Year: 2013

RATIONALE Reported incidents of the use of nutritional supplements containing deer antler velvet by athletes has increased significantly in recent years. The supplements have been reported to contain insulin-like growth factor-1 (IGF-1), which is a banned substance included on the World Anti-Doping Agency (WADA) prohibited list. The presence of deer and human IGF-1 was tested in six commercially available supplements. METHODS IGF-1 was extracted from the six deer antler velvet supplements using chloroform and acetonitrile precipitation methods. Ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) methods were developed to measure intact IGF-1 protein and IGF-1 trypsin peptides using a triple quadrupole mass spectrometer. Five deer-specific and five human-specific multiple-reaction monitoring (MRM) transitions for intact IGF-1were measured as well as six deer-specific and seven human-specific MRM transitions for an IGF-1 trypsin peptide. RESULTS The peak area from each MRM transition was used to calculate the product ion ratios relative to the most abundant transition. Product ion ratios measured in the supplements were matched to ratios measured in purified protein standards. A match to human IGF-1 was identified for all the MRM transitions measured in four of the supplements tested. CONCLUSIONS The presence of a pharmaceutical protein, human IGF-1, was confirmed in four commercially available products sold as all natural, nutritional supplements. These methods can be used to screen additional products to further prevent the illegal sale of adulterated supplements. Copyright © 2013 John Wiley & Sons, Ltd.

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