Technische Universität München

The Entrepreneurial University

 
All samples are solely analysed by WADA accredited laboratories (only 33 laboratories worldwide). WADA accredited laboratories obligatory have to use The World Anti-Doping Code, an International Standard for Laboratories. It constitutes a level 2 mandatory International Standard developed as part of the World Anti-Doping Program. The equipment is stipulated as follows:  

Gas chromatography (GC) with mass spectrometry (MS) and tandem mass spectrometry (MS/MS)

... for detecting stimulants, narcotics, diuretics, steroids, beta-blockers, etc.

A gas chromatograph is used to separate volatile compounds. The separated compounds are guided into the ion source (on-line) of a mass spectrometer (detector), which consists of a metallic filament with high voltage applied.. The compounds are ionized and their mass-to-charge ratio of intact ions detected (MS). Using a tandem mass spectrometer the compounds can be fragmented, yielding predictable patterns. These patterns can be used for enhanced identification of the compound (precursor-ion) using MS/MS.  

Liquid chromatography (LC) with detection via UV/VIS- or mass spectrometry (MS) and tandem mass spectrometry (MS/MS)

... for detecting glucocorticosteroids, stimulants, narcotics, diuretics, etc.

Liquid chromatography separates soluble compounds chromatographically before they are analysed in the detector. This is either done by their absorbance (UV/VIS detector) or by their mass-to-charge ratio (mass spectrometer). The method differs from GC/MS in that the mobile phase is liquid, usually a combination of water and organic solvents, instead of gas.  

Gas chromatography with combustion and isotope ratio mass spectrometry (GC/C/IRMS)

... to distinguish endogenous and exogenous steroids.

Isotope ratio mass spectrometry (IRMS) is a specialization of mass spectrometry, in which mass spectrometric methods are used to measure the relative abundance of isotopes in a given sample. The isotope ratio mass spectrometer allows the precise measurement of mixtures of stable isotopes. It is much more precise than a conventional spectrometer because measurements are repeated many times. The dual inlets of the instrument enable reliable repetition of measurements by supplying a continuous stream of reference and sample gases which are sequentially switched by a changeover valve. One type of  IRMS's collector also features an array of Faraday cups (conductive, metal vessels which neutralize ions that hit them whilst themselves becoming charged), the "multicollector" allows the simultaneous detection of multiple isotopes.
Samples must be introduced in the gas phase, achieved through combustion, gas chromatographic feeds or chemical trapping. By comparing the detected isotopic ratios to a measured standard, an accurate determination of the isotopic identification of the sample is obtained. For example, carbon isotope ratios are measured relative to the international standard for CO2, being fossil belemnite (found in the PeeDee formation, a limestone formed in the Cretaceous period in South Carolina, U.S.A.) with a 13C:12C ratio of 0.0112372. Because the difference between the reference and samples is often very small, the carbon isotope ratios are expressed as parts per thousand relative to the standard.  

Isoelectric Focusing, Immunoblotting and chemiluminescence detection

...for erythropoietin (EPO) analysis.

Isoelectric focusing (IEF) takes place in a pH gradient and can only be used for amphoteric substances such as peptides and proteins. The molecules move towards the anode or cathode until they reach a position in the pH gradient where their net charges are zero. This pH value is defined as  the “isoelectric point” (pI) of the substance. Being uncharged at this value, the electric field does not have any influence.
Blotting is the transfer of large molecules on to the surface of an immobilizing membrane. This method broadens the possibilities of detection for electrophoretically separated fractions because the molecules absorbed on the membrane surface are freely available for macromolecular ligands, for example antigens and antibodies.
The specific binding of antibodies is used to probe for individual protein zones and the signal is further increased by chemiluminescence, e.g. by binding a dye via the biotin-streptavidin peroxidase. To detect the signalbands, the membrane is exposed to an appropriate CCD camera in an absolutely dark closet for a certain time period.  

Hematological analyzer and cytometer

... for hematological parameters and for detecting blood transfusion.

Flow cytometry is a technique for counting, examining, and sorting microscopic particles suspended in a stream of fluid. It allows simultaneous multiparametric analysis of the physical and/or chemical characteristics of single cells flowing through an optical and/or electronic detection apparatus. Modern flow cytometers are able to analyze several thousand particles every second, in "real time", and can actively separate and isolate particles having specified properties. A flow cytometer is similar to a microscope, except that instead of producing an image of the cell, flow cytometry offers "high-throughput" (for a large number of cells) automated quantification according to a set of parameters.  

Immunoluminometric assays

... to distinguish pituitary and recombinant growth hormone (hGH).

An excess of two antigen-specific monoclonal antibodies that recognize the antigen (hGH) at different domains is used. The first antibody is immobilized on the inner surface of a tube. For first assay the coating antibody preferentially recognizes the recombinant hGH whereas for the second assay the coating antibody preferentially recognizes pituitary hGH. Secondary antibody is luminescence-labeled and is used in common for both assays. Luminescence is detected by using an automatic luminometer.  

Fluorometric assay method

... for detecting peptide hormones.

Fluorescence spectroscopy, also called fluorometry, or spectrofluorimetry is a type of electromagnetic spectroscopy which analyzes the fluorescence of a sample. The electrons of a molecule are excited by a beam of light, usually ultraviolet light. This causes the emission of light of a lower energy, typically, but not necessarily, visible light. A complementary technique is absorption spectroscopy. Devices that measure fluorescence are called fluorometers or fluorimeters.
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