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The marine environment is one of the most affected systems; especially the sediment compartment. Microplastic separation from the sediment matrix is the first step to evaluate its abundance and availability. Nevertheless, a lack of consistency in extraction protocols is a fact. This paper describes the optimization of the microplastic extraction procedure from marine sediments. The Plackett-Burman saturated factorial design was used to identify the significant factors and to select optimum working conditions. With this purpose, the following variables were studied: the number of extractions; the amount of sediment; the settling time; the density separation solution volume; the agitation time and the suitability of using wet or freeze-dried sediment. The Plackett-Burman design has revealed that the most statistically significant variables were sediment mass and agitation time. 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