Common ragweed (Ambrosia artemisiifolia L.) is an annual Asteraceae species native to North America which is highly invasive across Europe and has harmful impacts, especially on human health and agriculture. Besides its wide ecological range, particularly its high reproductive power by seeds is promoting its spread to various habitats and regions. To prevent further spread and to control the plant, the European Commission funded projects and COST-Actions involving scientists from all over Europe. A joint trial was set up comprising eight different laboratories from Europe to study seed viability variation in different seed samples. Three different testing methods (viability test with 2,3,5-triphenyltetrazolium chloride (TTC), a germination test combined with a subsequent TTC test and a crush test) were tested within the EU-COST-Action SMARTER network to four different seed origins. The viability test results from different laboratories were compared for variation amongst tests and laboratories. The main aim was to optimise the reliability of testing procedures, but results revealed not only significant effects of seed origin and seed age on seed viability, but also considerable differences between the output of the individual testing methods and furthermore between laboratories. Due to these significant differences in the results of the testing labs, additionally a second test was set up. Twelve Austrian ragweed populations were used for TTC testing to obtain a precise adjustment of the testing method as well as a tight guideline for interpreting the results, particularly for the TTC state “intermediate” since a proper classification of TTC-intermediate coloured seeds is still a challenge when determining viability rates.