Prof. Dr. W. Schänzer of the Cologne lab has penned a response to the EPO study released the other day in the Journal of Applied Physiology that called into question the urine test for EPO.

The World Anti-Doping Agency e-mailed 10 Speed a copy of the letter, which Prof. Dr. Schänzer sent to the editor of the Journal of Applied Physiology. Prof. Dr. Schänzer didn’t mince words in addressing the study: “The results of the publication are factually wrong, demonstrate great ignorance regarding the criticised method and the operations of doping control laboratories, and are based on a serious lack of careful examination of provided data. This is scientifically unacceptable.”

Further, he stated: “The central statement of the study is wrong. … The main result of the study is factually wrong and delusive. … The publication of data without thoughtful and critical review is scientifically not acceptable. … The great lack of knowledge of the authors was not known to laboratory B.”

Again, ol’ 10 Speed is not a scientist, nor do I play one on TV or this blog, so for the “Bill Nye the Science Guy” fans out there, here’s the Prof’s letter …

Comment of Laboratory B on the publication by Lundby et al. “Testing for recombinant human erythropoietin in urine: problems associated with current anti doping testing. J Appl Physiol 2008, doi :10.1152/japplphysiol.90529.2008″

The results of the publication are factually wrong, demonstrate great ignorance regarding the criticised method and the operations of doping control laboratories, and are based on a serious lack of careful examination of provided data. This is scientifically unacceptable.

In May 2007, laboratory B agreed to participate in a research project supported by
the Danish Antidoping Agency (Anti-Doping Denmark), which aimed, amongst others, to test for the traceability of rHuEpo when Lorzathan was co-administered. Before acceptance of the invitation, laboratory B requested assurance that the study was approved by an anti-doping organisation (Anti-Doping Denmark) and that the samples were not originating from athletes.

At no point, laboratory B was informed about the goal of an interlaboratory
comparison with regard to the analysis of rHuEpo with subsequent publication of the acquired data. After laboratory B found out about the publication, all data were
screened and evaluated again. - In the following are the comments of laboratory B:

The central statement of the study is wrong

The quality of the current detection method for rHuEpo (IEF-method) can not be
judged by the authors as the results, which were provided by laboratory B, were not obtained by means of the accredited method for urine analysis.

1. All samples were measured using the criticised IEF-methodi as well as a novel
approach based on SDS-PAGEii. The results reported to the authors were based on
SDS-PAGE analyses only. This is documented and verifiable. Laboratory B was
interested in the application of the SDS-PAGE method to administration study urine samples.

2. Laboratory B did not apply the ENTIRE IEF-method for the detection of rHuEpo that is commonly used for doping control purposes and was criticised by the authors. Only a SCREENING analysis was performed. The entire method includes three steps:

1. Screening analysis, 2. Repetition of analysis including different tests for activities that can yield in false positive results, 3. Inspection of results by a second laboratory. Steps 2 and 3 were not conducted. Hence, the quality of the current method (IEFmethod) can not be judged by the authors. The use of the abbreviated screening approach is also major reason why no negative or positive (adverse analytical finding) results were reported, but only unconfirmed SUSPICIOUS test results.

If the authors would have requested a statement regarding the differing results obtained by laboratories A and B or a proof-reading of the manuscript, all misunderstandings would have been eliminated immediately.

The main result of the study is factually wrong and delusive

The authors claim in the abstract that laboratory B did not determine any misuse of rHuEpo. This statement is wrong and misleading. Laboratory B did not make this comment, and the provided data do not allow the authors to draw such a conclusion. Laboratory B reported 15 suspicious samples; i.e., 15 specimens fulfilled screening criteria for the PRESENCE of rHuEpo and, thus, were SUSPICIOUS in terms of doping control practices. Confirmatory analyses were not conducted. Those would have verified which of the tested samples were positive and which were suspicious due to possible activities.Consequently, the main result of the publication (laboratory A detected misuse of Epo, laboratory B did not detect misuse of Epo) is outright false.

The data of laboratory B were not questioned by the authors

As stated above, laboratory B reported 15 samples as suspicious. The authors never asked how or why laboratory A obtained seriously deviating results. Moreover, they never asked what the term “suspicious” means. The fact that these results were obtained from screening analyses, which fulfil the criteria for adverse analytical findings but require further analyses, would have been readily explained to the authors. Such questions would have been asked by any sport federation if such a result was reported on athletes’ urine samples. It was the laboratory B’s understanding that the authors understood the analysis report and did not
provide any further details.

The publication of data without thoughtful and critical review is scientifically not acceptable.

The authors and reviewers/advisors should have known that the results of the
analyses were not reported by laboratory B as doping control analyses
The analysis report of laboratory B was not in accordance to the common guidelines for doping control samples. Such reports contain either negative, adverse, or atypical analytical findings. The term “suspicious” is not used in a doping control analysis report. The particular wording was used intentionally by laboratory B to outline the fact that no final decision about a positive or negative result was made. Obviously the authors (and reviewers/advisors) were not familiar with rules and wordings commonly applied in doping control laboratories. This ignorance has led to wrong interpretations of results and caused a serious damage to the reputation of the rHuEpo detection method, the doping control system as well as laboratory
B.

Laboratory B was not informed about the goal of the study

The intention of the authors to test the performance of different laboratories was not mentioned at any time. If laboratory B would have known about this aim, a participation would have been declined as the test was not conducted under controlled conditions and by using certified test material. Moreover, laboratory B was not informed about the publication of the data, and the manuscript was never provided for proof-reading. The false interpretation of the results and the factually wrong presentation of data would have been prevented

The operations in laboratory B were not defined

As the authors have obviously not been aware of the operations being conducted in doping control analyses (screening analysis and subsequent confirmation in case of suspicious results), a request for confirmations of suspicious samples was not made. Laboratory B used SDS-PAGE for screening purposes only, and outlined this fact by reporting SUSPICIOUS results only instead of negative or adverse/atypical analytical findings. Detailed evaluations of borderline case samples were not conducted. The great lack of knowledge of the authors was not known to laboratory B.

i Lasne F, Martin L, Crepin N, de Ceaurriz J: Detection of isoelectric profiles of erythropoietin in urine: differentiation of natural and administered recombinant hormones. Anal Biochem. 311 (2002) 119-26

ii Kohler M, Ayotte C, Desharnais P, Flenker U, Lüdke S, Thevis M, Völker-Schänzer E, Schänzer W: Discrimination of recombinant and endogenous urinary erythropoietin by calculating relative mobility values
from SDS gels. Int J Sports Med 29 (2008) 1-6

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