The purpose of this study was to evaluate the effects of sodium hypochlorite on the organic and inorganic composition of enamel. Background: With the advent of enamel bonding for orthodontic appliances in the late 1970s, it has been shown that traditional phosphoric acid etching affects the inorganic portion of the enamel.1, 2 In an attempt to enhance the acid etching pattern and, furthermore, the bond strength, additional pretreatment techniques that target the organic components of the enamel biofilm have been proposed. One such method is the non-invasive enamel pretreatment with 5.25% sodium hypochlorite (NaOCl) prior to phosphoric acid etching.3, 4 It has been suggested that the mechanism by which sodium hypochlorite enhances the etching pattern is enamel deproteinization, in which organic elements, including the acquired film, are removed from the enamel surface.3, 5 This presumption is based on the multitude of endodontic literature supporting the use of NaOCl as an effective irrigant in root canal therapy6-13. In contrast to dentin and pulpal tissue, enamel is comprised of minimal organic matter.14, 15 As a result of this fact and the limited amount of experimentation of the effect of NaOCl on the enamel surface, the true mechanism by which sodium hypochlorite enhances the etching pattern of enamel is questionable.5, 16 The objective of this study was to determine the compositional effects of sodium hypochlorite on human enamel. Methods: Following IRB approval, 120 enamel sections from 22 extracted human premolar teeth were randomly divided into three experimental groups and one control group.17 The control group (E = enamel) received no treatment. The first experimental group (A = phosphoric acid) received a 15-second treatment with 37% phosphoric acid, rinsed with distilled water and air sprayed for 20 seconds, then dried with oil free compressed air. The second experimental group (H = sodium hypochlorite) received a treatment of 5.25% sodium hypochlorite for 60 seconds, washed with distilled water for 10 seconds, and dried. The third experimental group (HA = sodium hypochlorite + phosphoric acid) received a treatment of 5.25% sodium hypochlorite for 60 seconds, washed with distilled water for 10 seconds, dried, then receive the 15-second treatment with 37% phosphoric acid as in Group A.3 Following treatment preparations of the four groups, scanning electron microscopy (SEM)/energy-dispersive X-ray spectrometer (EDX) analysis was performed for all groups.18 For elemental concentration, a one-way ANOVA and Tukey’s post hoc statistical tests were applied.17, 19, 20 ANOVA and Tukey tests were performed at a significance level of p ≤ 0.05. Results: There were no significant effects of treatment on the enamel elements carbon (C), calcium (Ca) sodium (Na), oxygen (O), and phosphorous (P). There was a significant effect of treatment on the amount of chlorine (Cl) in enamel between groups acid (A) and hypochlorite + acid (HA) as well as between groups hypochlorite (H) and hypochlorite + acid (HA) (p = 0.004). The amount of variation of iodine (I) in the enamel composition between untreated enamel (E) and enamel treated with sodium hypochlorite + phosphoric acid (HA) was significant (p = 0.004). Additionally, there was a significant decrease in the quantity of antimony (Sb) found in the control group (E) versus the hypochlorite + acid (HA) experimental group (p = 0.002). Lastly, tin (Sn) was significantly reduced from the enamel surface (E) when treated with hypochlorite + acid (HA) (p = 0.008). Conclusions: The various treatments minimally affected the elemental concentrations of C, Ca, Na, O, and P. The amount of chlorine present in enamel significantly increased following treatment with sodium hypochlorite (H) alone and even more so following treatment with phosphoric acid and sodium hypochlorite (AH). In contrast, elements I, Sb, and Sn demonstrated a congruent reduction in concentration after treatment with hypochlorite and acid (HA). Although it has been hypothesized that sodium hypochlorite targets the organic pellicle present on the surface of enamel via a process known as deproteinization, the findings presented here suggest that pre-treatment with NaOCl impacts the inorganic components of enamel more so than the organic constituents. These quantitative findings corroborate the enhanced etching pattern that can be visualized under scanning electron microscopy in this as well as previous studies