基于特征图谱和含量测定的乌梅丸质量标准研究

Quality Standard of Wumei Pills Based on Characteristic Chromatograms and Content Determination

目的建立乌梅丸的质量控制方法。方法采用中药色谱指纹图谱特征图谱,结合高效液相色谱(HPLC)法测定乌梅丸(散剂)及3批药典方乌梅丸(大蜜丸)中枸橼酸、盐酸黄柏碱、党参炔苷、盐酸黄连碱、羟基-α-山椒素指标性成分的含量。色谱柱为Waters Symmetry Shield C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为0.8 mL/min,检测波长为214 nm(枸橼酸)和285 nm(其余4种成分),柱温为27℃,进样量为10μL。结果通过乌梅丸(散剂)特征图谱确定共有峰为22个特征峰。枸橼酸、盐酸黄柏碱、盐酸黄连碱、党参炔苷、羟基-α-山椒素的进样量分别在0.278~16.704μg、0.010~0.197μg、0.048~0.956μg、0.269~5.386μg、1.841~36.818μg范围内与峰面积线性关系良好,r分别为0.9999,0.9999,0.9998,0.9996,1.0000;加样回收率分别为103.90%,95.62%,97.23%,96.74%,100.24%,RSD分别为1.04%,2.71%,2.14%,5.48%,5.69%(n=9);精密度、稳定性、重复性试验的RSD均小于3.0%。乌梅丸(散剂)中枸橼酸、盐酸黄柏碱、盐酸黄连碱、羟基-α-山椒素含量的平均值分别为87.22,0.43,3.56,1.86 mg/g,党参炔苷的含量最低(0.09 mg/g);乌梅丸(大蜜丸)不含党参炔苷,枸橼酸、盐酸黄柏碱、盐酸黄连碱、羟基-α-山椒素含量的平均值分别为142.85,0.25,2.54,0.20 mg/g。结论所建立的方法操作简便、准确、重复性好,可用于乌梅丸的质量控制。

Objective To establish a quality control method of Wumei Pills.Methods The characteristic chromatograms of chromatographic fingerprint of traditional Chinese medicine(TCM)and high-performance liquid chromatography(HPLC)method were used to determine index components such as citric acid,phellodendrine chloride,lobetyolin,coptisine hydrochloride and hydroxy-α-sanshool in Wumei Pills(Power)and three batches of Wumei Pills(Dami Pills)prepared according to the Chinese Pharmacopoeia.The chromatographic column was Waters Symmetry Shield C_(18) column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.1%phosphoric acid solution(gradient elution),the flow rate was 0.8 mL/min,the detection wavelength was set at 214 nm for citric acida and 285 nm for the other four kinds of components,the column temperature was 27℃and the injection volume was 10μL.Results Twenty-two common characteristic peaks were identified in HPLC characteristic chromatograms of Wumei Pills(Power).The linear ranges of citric acid,phellodendrine chloride,coptisine hydrochloride,lobetyolin and hydroxy-α-sanshool were 0.278-16.704μg,0.010-0.197μg,0.048-0.956μg,0.269-5.386μg and 1.841-36.818μg(r=0.9999,0.9999,0.9998,0.9996 and 1.0000).The average recoveries of the above components were 103.90%,95.62%,97.23%,96.74%and 100.24%with RSDs of 1.04%,2.71%,2.14%,5.48%and 5.69%(n=9).The RSDs of precision,stability and repeatability tests were less than 3.0%.The average contents of citric acid,phellodendrine chloride,coptisine hydrochloride and hydroxy-α-sanshool in Wumei Pills(Power)were 87.22,0.43,3.56,1.86 mg/g,and the content of lobetyolin in Wumei Pills(power)was the lowest(0.09 mg/g).There was no lobetyolin in Wumei Pills(Dami Pills),while the average contents of citric acid,phellodendrine chloride,coptisine hydrochloride and hydroxy-α-sanshool in Wumei Pills(Dami Pills)were 142.85,0.25,2.54 and 0.20 mg/g.Conclusion The established method is simple,accurate and reproducible,which can be used for the quality control of Wumei Pills.