The introduction of new bioanalytical techniques to assay blood samples as well as the application of modified blood sampling techniques at NOTOX has led to a 40% reduction in test animal usage needed in standard kinetic PK studies. “This is again one important step forwards in our endeavor to respond to the 3R’s principle: “Reduction, Refinement and Replacement” Mira Wenker, Head of DMPK at NOTOX, says. “It is simply a result of our internal discussions and bringing the right people from different sciences together”.

Bioanalytical Study Director Michael Eman: “It all started with the combination of UPLC and the API 5000 MSMS in our lab, a very powerful tool for the fast and very sensitive analysis of pharmaceuticals in plasma. This combination requires much less blood sample than conventional LC/MS analysis, while still the same or better detection limits apply.” “Our main challenge was the handling of much smaller volumes of liquid samples” Michael continues, “10 instead of 50 μl of plasma and extracts and still preserving the high standards of accuracy and repeatability. Finally we got hold of a suitable procedure.” “Normally we drew 300 μl blood from the orbita (venous  eye plexus) from a rat for each PK/TK time point, and for 8 time points under anaesthesia within 24h this means that more than 1 rat is needed to construct the entire curve” Mira adds. “With the 100 μl blood we need now, we can draw 8 times from the tail-vein of the same rat and no anesthesia is necessary. This means less discomfort for the animal and also less animals, thus hitting 2 out of the 3 R’s: refinement and reduction. And for the benefit of science: we can now construct our TK/PK curves from one animal, leading to a better interpretation of the results.”

See the flyer below to learn more about this innovative technique.