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Ginseng is a well-known traditional Chinese medicinal herb, and ginsenosides are its major active components. A method for the fast determination of ginsenosides in ginseng samples by high-performance liquid chromatography was developed and used for the quantitative analysis of four ginsenosides in three different ginseng samples. In this method, instead of time-consuming gradient elution, isocratic elution was used to speed up the analysis. Under strong isocratic elution, all the ginsenosides are eluted in 2.3 min. Although the measured signal is composed of overlapped peaks with the interferences and background, the signal of ginsenosides can be extracted by chemometric resolution. A non-negative immune algorithm was employed to obtain the chromatographic information of the target components from the data. Compared with conventional chemometric approaches, the method can perform the extraction for one-dimensional overlapping signals. The method was validated by the determination of four ginsenosides in three different ginseng samples. The recoveries of the spiked samples were in the range of 94.08-107.3%. [ABSTRACT FROM AUTHOR]