Retention of active ingredients of toothpastes in the mouth following brushing determines the efficiency of these oral care formulations. In this study, new in vitro methodologies for the observation and measurement of toothpaste retention in the oral cavity were developed and used to evaluate the efficiency of formulations containing different mucoadhesive hydrophilic polymers. The findings suggest that using Carbopol ETD 2020 and Carbopol Ultrez 10 as binders in toothpaste prolongs the retention time of these formulations in the oral cavity. The in vitro methodologies tested, coupled with texture analysis, were able to accurately characterise the behaviour of the toothpaste and produce detailed images showing how it is retained in the oral cavity. This study has not only produced a new method for studying the behaviour of toothpaste and other formulations in the oral cavity but is also the first to investigate how different types of mucoadhesive binders can be used to improve toothpaste retention.