[Dual-wavelength method for analysis of overlapped high performance liquid chromatographic peaks with multiwavelength...

High performance liquid chromatography (HPLC) is one of the analytical techniques most frequently used. Its chief limitation lies in the need to avoid the overlap of chromatographic peaks as much as possible. In this work, a new method for the treatment of HPLC data allowing the resolution of extensively overlapped peaks has been developed. This method uses a diode array spectrophotometer and treats the generated data by dual-wavelength method. Its foundation is based on calculating the peak areas for two given wavelength in order to determine the contribution of each component to the mixed peak. Because the area is the integration of absorption at different wavelength, the wavelength can be chosen from the absorption spectra of the two compounds for eliminating the interference of the other component in the mixture, and the concentration of the corresponding component can be calculated. This method provides a simplification of the calculation process and a reduction of the amount of the data used. The applicability of the method is checked on mixture of two aromatic nitro-compounds. Though the two components almost have the same retention time and their peaks overlapped extensively, the results are satisfactory.