Objective: The objective of this in vitro study was to assess the effect of several chelating agents on the mineral content of root dentin. Study design: Extracted human mandibular incisor roots were prepared and divided into groups according to the following irrigation protocols: 1) 17% ethylenediaminetetraacetic acid (EDTA); 2) 10% citric acid solution; 3) 18% etidronate; 4) 2.25% peracetic acid; 5) and deionized water (control). Dentin chips were obtained (Gates-Glidden nos. 3, 4, and 5). The levels of different minerals were analyzed with the use of inductively coupled plasma-atomic emission spectrometry (ICP-AES). Results: 1) Peracetic acid significantly decreased P, K, Mg, Na, and S levels compared with the other groups (P <.05). 2) S decreased by different levels in all of the chelating solutions (P <.05), and the greatest decrease was observed in peracetic acid. 3) Ca levels significantly decreased in peracetic acid, citric acid, and EDTA (P <.05). 4) Mn levels significantly decreased in the citric acid and peracetic acid groups (P <.05). 5) Na and Zn levels significantly decreased in the peracetic acid, citric acid, and etidronate groups (P <.05). Conclusions: The chelation agents can create different effects on mineral contents of root dentin, so it is important to know what effects each solution will have on root dentin before their clinical use. In addition, according to the results of this in vitro study, it might be recommended that peracetic acid, in particular, should be used with caution. © 2011 Mosby, Inc. All rights reserved.