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Triclosan (TCS) is used in many health care products as a broad-spectrum antimicrobial and disinfectant agent; however, it is currently considered an emerging pollutant. The present study aimed to develop an analytical HPLC method coupled to a Diode Array Detector for the TCS determination in toothpaste. Method development involved the optimization of separation and sample preparation. For chromatographic separation optimization, different RP columns and aqueous mobile phases were addressed. The optimized chromatographic separation was achieved in around 3 minutes using a Symmetry C18 column at 30 °C, 0.6 mL/min flow-rate, and 225 nm wavelength detection. For toothpaste sample preparation optimization a response surface methodology using a Box-Behnken fractional factorial design was chosen. The sample treatment consisted of leaching 0.2 g sample by sonication for 7 minutes using 2 mL of MeOH. According to the International Conference on Harmonization (ICH, 1996/2005) and European Commission Decision 200/657/EC guidelines, the developed method was validated. The developed analytical method was found fast, economic, linear, sensitive, precise, and robust and allowed to quantify TCS in different healthcare products.