土一枝蒿药材指纹图谱和含量测定研究

Fingerprints and Content Determination of Achillea Wilsoniana

目的研究土一枝蒿药材的指纹图谱和相关成分含量测定方法。方法参照2020年版《中国药典(四部)》测定药材样品的水分、总灰分、水溶性浸出物含量;采用高效液相色谱(HPLC)法测定药材样品中3个成分的含量,色谱柱为CAPCELL PAK C18MGⅡ(S-5)柱(250 mm×4.6mm,5μm),流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为0.7mL/min,检测波长为365 nm,柱温为30℃,进样量为10μL。建立10批药材样品的HPLC指纹图谱,采用中药色谱指纹图谱相似度评价系统(2012年130723版)进行相似度评价,确定共有峰。结果10批药材样品的HPLC图谱有14个共有峰,相似度为0.941~0.997,确认的3个成分分别为异荭草素、异牡荆苷、槲皮素-4′-O-β-D-葡萄糖苷,其质量浓度分别在12.55~200.82μg/mL(r=0.9999)、7.20~115.28μg/mL(r=0.9995)、6.14~98.20μg/mL(r=0.9998)范围内与峰面积线性关系良好;精密度、稳定性、重复性试验结果的RSD均小于3.0%;平均加样回收率分别为97.92%,101.37%,103.92%,RSD分别为1.38%,2.33%,1.26%(n=6)。10批药材样品中上述3个成分的平均含量分别为1.4210,0.4220,0.2834mg/g;水分含量为10.15%~12.23%,总灰分含量为5.24%~13.29%,水溶性浸出物含量为7.37%~11.54%。结论所建标准可用于土一枝蒿药材的质量控制。

Objective To study the fingerprints of Achillea wilsoniana,and to establish a method for the content determination of related components.Methods According to the Chinese Pharmacopoeia(2020 Edition,VolumeⅣ),the content of moisture,total ash and water-soluble extracts of the medicinal materials were determined.The content of three components in the medicinal materials was determined by the high-performance liquid chromatography(HPLC)method with CAPCELL PAK C18MGⅡ(S-5)column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-0.1% phosphoric acid solution(gradient elution),the flow rate was 0.7mL/min,the detection wavelength was 365 nm,the column temperature was 30℃,and the injection volume was 10μL.HPLC fingerprints of 10 batches of medicinal materials were established to evaluate the similarity and determine the common peaks by the Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine(Version130723,2012).Results There were 14 common peaks in the HPLC chromatograms of 10 batches of medicinal materials,and the similarity was in the range of 0.941-0.997.The three components identified were isoorientin,isovitexin,and quercetin-4′-O-β-D-glucoside,and their linear ranges were 12.55-200.82μg/mL(r=0.9999),7.20-115.28μg/mL(r=0.9995),6.14-98.20μg/mL(r=0.9998).The RSDs of precision,stability and repeatability tests were all less than 3.0%.The average recoveries of isoorientin,isovitexin,and quercetin-4′-O-β-D-glucoside were 97.92%,101.37%,103.92% with RSDs of 1.38%,2.33% and 1.26%(n=6),respectively.The average content of the above three components in 10 batches of medicinal materials were 1.4210,0.4220,0.2834mg/g,respectively.The moisture content was in the range of 10.15%-12.23%,the total ash content was in the range of 5.24%-13.29%,and the water-soluble extract content was in the range of 7.37%-11.54%.Conclusion The established standard can be used for the quality control of Achillea wilsoniana.