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Herbal medicine (HM) formulae are the combinations of two or more types of constituting herbs. This study has proposed a novel approach to efficiently develop HPLC methods for HM formulae, which take advantage of the mutual retention rules between HM formulae and their constituting herbs. An HM formula composed of two herbs, Radix Salviae Miltiorrhizae and Rhizoma Chuanxiong, was taken as a case study. Based on design of experiments and stepwise multiple linear regression, models relating the analytical parameters to the chromatographic parameters were built (correlation coefficients &gt;0.9870) for chemical compounds in the two herbs. These models representing the retention rules were utilized to predict the elution profile of the formula. The analytical parameters were numerically optimized to ensure adequate separation of the analytes. In validation experiments, satisfactory separations were achieved without any pre-experiments on the formula. The approach can significantly increase the HPLC method development efficiency for HM formulae.

Rapid HPLC Analytical Method Development for Herbal Medicine Formulae Based on Retention Rules Acquired from the Constituting Herbs