BACKGROUND: The surge of studies on artificial and natural nanoparticles has revealed a new world for engineering and life sciences, but in most instances, the small scale has made their number-concentration determination in liquids a challenging problem. Former success has mostly been limited to special particles measured by indirect techniques. A new approach is required for this determination to facilitate the production and application of nanoparticles in different systems. RESULT: Here, an approach is described using a nanoparticle tracking system based on Brownian motion, which can be used to determine the number-concentration of nanoparticles, including viruses, in liquids on-line. Extensive analysis showed the influence of suspension concentration and particle size on the accuracy of measurements. Natural nanoparticles of Adenovirus and several types of artificial nanoparticles, including precision nanobeads, uniform inorganic silica microspheres, monodisperse gold metal colloids and aggregated Aerosil nanoparticles, were measured and compared by counting the monitored particle number obtained using light scattered from individual particles, from which the particle number-concentration, the product yield and the aggregation could be evaluated. CONCLUSION: This approach was compared with the mathematical calculation method and the emission spectrophotometry technique used for practical applications. The results showed this new approach had improved accuracy for determination of the particle number-concentration. (C) 2010 Society of Chemical Industry