Multiple Compounds Determination Of Zhizi-jinhua Pills By High-performance Liquid Chromatography

Objective: The aim of this study was to establish a method for simultaneously determinating the contents of multiple marker compounds in Zhizi-jinhua pills by highperformance liquid chromatography(HPLC) for comprehensively controlling the quality of it.Methods: All the separations were carried out on the ZORBAX Eclipse XDB-C18 column(250×4.6 mm, i.d., 5 μm). The mobile phase consisted of acetonitrile(A) and water(contain 0.3% phosphoric acid and 0.2% triethylamine, B) in a gradient elution mode was used as follows: 0 min, 5%A; 10 min, 15%A; 70 min, 30%A; 85 min, 30%A; 105 min, 60%A; 120 min, 60%A. The flow rate was 1.0 mL/min, and detection wavelength was set at 270 nm in room temperature. At the same time, the methodologies for determination of geniposide, baicalin, quercetin, palmatine, berberine, wogonoside, baicalein, emodin and chrysophanol including specificity, standard curve, limit of detection(LOD), limit of quantification(LOQ), precision, repeatability, stability and recoveries were tested. And then, the contents of the nine marker compounds in Zhizi-jinhua pills collected from different companies with different batches were assayed.Results: Under the selected conditions, the compounds of geniposide, baicalin, quercetin, palmatine, berberine, wogonoside, baicalein, emodin and chrysophanol achieved base separation with no interferences produced by other chemicals in Zhizijinhua pills.. Good calibration curves of the markers in the ranges of 30.63~1960.32 μg/m L(R2 = 0.9999), 9.38~600.32 μg/mL(R2 = 0.9999), 0.82~52.48 μg/m L(R2 =0.9999), 3.60~230.40 μg/m L(R2 = 0.9997), 3.99 ~255.36 μg/m L(R2 = 0.9997), 7.04~450.56 μg/m L(R2 = 0.9999), 15.12~999.68 μg/m L(R2 = 0.9999), 4.06~259.84 μg/m L(R2 = 0.9999) and 9.41~602.24 μg/m L(R2 = 0.9999), respectively, were produced. The relative standard deviation(RSD) of each compound was in the range of 0.22-2.38% for intra-day precision and 0.13-2.45% for inter-day precision assays. Good repeatability(RSD in the range of 0.64-2.03%) and stability(18 h in room temperature with RSD from 0.74 to 2.55%) were found. The recoveries of the compounds were in the range of 90.85-110.95% with RSD from 0.67 to 3.23%. The contents of geniposide were the highest, which were from 4693.32 to 11881.18 μg/g in all of the samples, and the contents of quercetin were the lowest from 24.49 to 58.24 μg/g. The results showed that the contents of the same compound were difference among the zhizi-jinhua pills from different manufactures, but the differences was little in the samples from the same manufacture.Conclusion: In this study, the HPLC method for simultaneoussly determinating the contents of geniposide, baicalin, quercetin, palmatine, berberine, wogonoside, baicalein, emodin and chrysophanol in Zhizi-jinhua pills was established, which was simple, rapid, and suitable for the quality control of Zhizi-jinhua pills.