Development and Validation of a HPLC Method for the Determination of Five Bioactive Compounds in the “Xuebijing” Injection

A reversed phase high performance liquid chromatographic (RP-HPLC) method with ultraviolet (UV) detector was developed for simultaneously determining five bioactive components (i.e., hydroxysafflor yellow A: HSYA; paeoniflorin, ferulic acid, benzoic acid, and danshensu) in “Xuebijing” (XBJ) injection, a widely used traditional Chinese medicine (TCM) for treating sepsis and multiple organ dysfunction syndrome. A Zorbox SB C18 column was used with 0.2% phosphoric acid (V/V)-acetonitrile as the mobile phase under the condition of gradient elution. The five components were analyzed by using a timed wavelength measure according to their maximum absorption wavelength. The intraday and interday precisions of the five investigated compounds were less than 1.17% and the average recoveries ranged from 97.3% to 103.2%. There were good linear correlations between the concentrations of the five components and their chromatographic peak areas (R2 ≥ 0.9998), the proposed method was successfully applied to determine the five components in different batches of XBJ injection products, the results indicated that the proposed method is simple, stable, and accurate and could be readily utilized as a quality control method for manufacturing process of XBJ injection.