N-Acetyl-L-cysteine (NAC) can inhibit the luminol–H2O2, reaction, which is catalyzed by silver nanoparticles. Based on this phenomenon a new method was developed for NAC determination. Under optimum conditions, a linear relationship between chemiluminescence intensity and NAC concentration was found in the range 0.034–0.98 μg/mL. The detection limit was 0.010 μg/mL (S/N =3), and the relative standard deviation (RSD) was <5% for 0.480 μg/mL NAC (n =5). This simple, sensitive and inexpensive method has been applied to measure the concentration of NAC in pharmaceutical tablets. Copyright © 2014 John Wiley & Sons, Ltd.