广西医学

目的建立同步测定药对大黄-黄芪胶囊主要蒽醌类成分含量的高效液相色谱法（HPLC）。方法采用HPLC检测药对大黄-黄芪胶囊中芦荟大黄素、大黄酸、大黄素和大黄酚的含量。色谱条件：Supersil ODS2色谱柱(250 mm×4.6 mm，5 μm)，柱温35 ℃，流动相为甲醇(A)-0.1%磷酸水溶液(B)，流速1.0 mL/min，梯度洗脱方式为0～70 min(50%～75%A)、70～90 min(75%A)，流速为1.0 mL/min，检测波长为254 nm。在上述色谱条件下进行专属性试验、线性关系考察、仪器精密度试验、稳定性试验、重复性试验、加样回收试验及样品含量测定。结果芦荟大黄素、大黄酸、大黄素和大黄酚的浓度分别为0.25～36.25 mg/mL(r=0.990 8)，0.50～53.50 mg/mL(r=0.994 2)、0.25～45.00 mg/mL(r=0.991 2)、0.50～62.00 mg/mL(r=0.993 7)，浓度与峰面积呈良好的线性关系。仪器精密度试验、稳定性试验、重复性试验的相对标准偏差（RSD）均＜3%。4种蒽醌类成分的平均回收率为98.97%～102.45%，RSD为0.26%～3.92%。芦荟大黄素、大黄酸、大黄素和大黄酚的含量分别为(0.821±0.062)mg/g、(1.753±0.159)mg/g、(1.303±0.109)mg/g、(2.909±0.251)mg/g。结论采用HPLC同步检测药对大黄-黄芪胶囊中芦荟大黄素、大黄酸、大黄素和大黄酚的含量，结果准确可靠、操作简便、重复性好，可用于大黄药材及其复方制剂药对大黄-黄芪胶囊主要蒽醌类成分的含量测定。

ObjectiveTo establish the high performance liquid chromatography (HPLC) for simultaneously determining contents of main anthraquinone components in drug-pair Radix rhei et rhizome-Astragalus membranaceus Capsule.MethodsThe contents of aloe emodin, rhein, emodin, and chrysophanic acid from drug-pair Radix rhei et rhizome-Astragalus membranaceus Capsule were determined by using HPLC. The chromatographic criteria were as follows: Supersil ODS2 chromatographic column was (250 mm×4.6 mm, 5 μm), column temperature was 35 ℃, mobile phase was methyl alcohol (A)-0.1% phosphoric acid aqueous solution (B), flow velocity was 1.0 mL/min, method of gradient elutions were 0-70 minutes (50%-75% A) and 70-90 minutes (75% A), flow velocity was 1.0 mL/min, detected wave strength was 254 nm. The specificity test, linear relation investigation, instrument precision test, stability test, repeatability test, sample recovery test, and sample assay were performed by the aforesaid chromatographic criteria.ResultsThe concentrations of aloe emodin, rhein, emodin and chrysophanic acid were 0.25-36.25 mg/mL (r=0.9908), 0.50-53.50 mg/mL (r=0.9942), 0.25-45.00 mg/mL (r=0.9912), 0.50-62.00 mg/mL (r=0.9937), and there was a favorable linear relation between concentrations and peak areas. The relative standard deviation (RSD) of instrument precision test, stability test, repeatability test were all less than 3%. The average recoveries of the 4 components of anthraquinone were 98.97%-102.45%, and RSDs were 0.26%-3.92%. The contents of aloe emodin, rhein, emodin, and chrysophanic acid were (0.821±0.062) mg/g, (1.753±0.159) mg/g, (1.303±0.109) mg/g, and (2.909±0.251) mg/g, respectively.ConclusionEmploying HPLC for simultaneously determining contents of aloe emodin, rhein, emodin, and chrysophanic acid from drug-pair Radix rhei et rhizome-Astragalus membranaceus Capsule exerts accurate and reliable results, easy and simple operation, with a favorable repeatability, which can be used for content determination of main anthraquinone components from medicinal material Radix rhei et rhizome and its compound preparation drug-pair Radix rhei et rhizome-Astragalus membranaceus Capsule.