Systematics is considered important for effective toothbrushing. A theoretical concept of systematics in toothbrushing and a validated index to quantify it using observational data is suggested. The index consists of three components: completeness (all areas of the dentition reached), isochronicity (all areas brushed equally long) and consistency (avoiding frequent alternations between areas). Toothbrushing should take a sufficient length of time; therefore, this parameter is part of the index value calculation. Quantitative data from video observations were used including the number of changes between areas, number of areas reached, absolute brushing time and brushing time per area. These data were fed into two algorithms that converted the behaviour into two index values (each with values between 0 and 1) and were summed as the Toothbrushing Systematics Index (TSI) value; 0 indicates completely unsystematic and 2 indicates perfectly systematic brushing. The index was developed using theoretical data. The data matrices revealed the highest values when all areas are reached and brushed equally long. Few changes occurred between the areas when the brushing duration was 90 s; the lowest values occurred under opposite conditions. Clinical applicability was tested with data from re-analysed videos from an earlier intervention study aiming to establish a pre-defined toothbrushing sequence. Subjects who fully adopted this sequence had a baseline TSI of 1.30±0.26, which increased to 1.74±0.09 after the intervention (p 0.001). When the participants who only partially adopted the sequence were included, the respective values were 1.25±0.27 and 1.69±0.14 (p 0.001). The suggested new TS-index can cover a variety of clinically meaningful variations of systematic brushing, validly quantifies the changes in toothbrushing systematics and has discriminative power.