The results of the analysis of the advantages and disadvantages of the use of retrospective data for the implementation of the evaluation of uncertainty in the measurement of indicators of biological safety of food and feed raw materials are given. The model measurement procedure used quantitative determination of genetically modified organisms by means of polymerase chain reaction in real time 250 measurement results of certified reference material with a content of 0.1% genetically modified soybean line obtained under conditions of intra-laboratory reproducibility and 10 results of relevant inter-laboratory comparisons with the participation accredited provider. The suitability of retrospective data was assessed by the Shapiro-Wilk criterion and statistical controllability analysis using standardized score charts and modified Shewhart charts, uncertainty by 4 methods described in general and branch international guidelines and regulations. The results of using a combination of modified Shewhart maps demonstrate the acceptability of this approach for establishing the suitability of retrospective measurement results as input data for uncertainty assessment. By integrating additional "fitness limits" to a set of standardized control limits, this analysis acquires the ability to simultaneously establish not only stability, through the interpretation of geometric patterns, but also compliance with specific criteria. Using this approach, objective evidence of the suitability of retrospective data for estimating measurement uncertainty was obtained. The distribution of retrospective data obtained under conditions of intralaboratory reproducibility is assumed to be normal. As a result of the assessment of statistical controllability and compliance with the established criteria, objective evidence of the suitability of the data for uncertainty assessment was obtained. Analysis of uncertainty intervals of subgroups with the volume of 25 measurements, using 4 assessment methods, demonstrated significant variability in the results of measuring biological safety indicators. The uncertainty interval could narrow or widen by almost 100% of the value within fifty measurements. Median values obtained using different uncertainty estimation methods were 2.69%, 22.69%, 39.23, and 43.08%. This variability is due to the limited coverage of sources of uncertainty, which is inherent in each of the analyzed methods. The most optimal method is the evaluation based on the routine measurement of the certified reference material. Retrospective results of routine measurement of certified reference material and interlaboratory comparisons are an effective source of input data for assessing the uncertainty of the results of measuring biological safety indicators of agricultural raw materials. There is a need for further research to establish an assessment model that will provide an opportunity to take into account the optimal number of influential sources of uncertainty that are inherent in biological systems and cannot be simultaneously taken into account using existing approaches offered by standard assessment methods.