Few studies have examined whether nicotine dependence self-report questionnaires can predict specific behaviors and symptoms at specific points in time. The present study used data from a randomized clinical trial (N = 608; M. E. Piper et al., 2007) to assess the construct validity of scales and items from 3 nicotine dependence measures: the Fagerstrom Test for Nicotine Dependence (T. F. Heatherton, L. T. Kozlowski, R. C. Frecker & K.-O. Fagerstrom, 1991), the Nicotine Dependence Syndrome Scale (S. Shiffman, A. J. Waters, & M. Hickcox, 2004), and the Wisconsin Inventory of Smoking Dependence Motives (M. E. Piper et al., 2004). Scales from these measures were used to predict participants' reports on real-time measures of withdrawal symptoms and smoking behavior and retrospective self-report questionnaires to assess convergent and discriminative validity. The nicotine dependence measures' scales and items generally predicted the real-time measures of similar constructs, but the percent of variance accounted for was low. The nicotine dependence measures did, however, show evidence of discriminative validity. Thus, this study provides modest support for the construct validity of these nicotine dependence scales. (Contains 10 footnotes, 5 tables, and 1 figure.)