One single standard substance for the determination of multiple anthraquinone derivatives in rhubarb using high-performance liquid chromatography-diode array detection

Due to the complexity of the chemical constitution of traditional Chinese medicines (TCMs), now there is a trend to establish methods, using multi-components analysis, for the effective quality control of TCMs. However, the limited availability of multiple reference substances hinders the wide popularization for routine quality control of TCMs. Using an easily available single component contained in the analyte as an internal standard to determine multiple analogues should be a practical option. In this study, we selected rhubarb rhizome as an example, and used emodin, the easily available active component, as the external standard to determine directly the content of emodin in rhubarb, and the same component as the internal standard to simultaneously determine the other six anthraquinones in rhubarb. Compared with the results obtained using the external standard method, this alternative method was found to have no statistically significant differences in our laboratory as verified by the F-test (p = 95%, n = 6). However, due to weak robustness caused by the fluctuation of relative response factors in different laboratories, such a method can only serve as a practical measure to solve the lack of so many chemical standard substances when relative response factors have been determined. This alternative method can then be used without reference substances. Once the corresponding chemical standards are available and are acceptable as well as cost-effective, then the external standard method for the simultaneous determination of multiple components (SDMC) in TCMs will prevail.