反枝苋药材的质量标准研究

Study on Quality Standard for Amaranthus retroflexus

目的:为反枝苋药材质量标准的制订提供参考。方法:以7批反枝苋药材为研究对象,对其进行外观性状及粉末特征显微观察;采用薄层色谱法(TLC)对反枝苋药材中芦丁、缬氨酸、亮氨酸进行定性鉴别,参照2015年版《中国药典》(四部)通则相关方法测定其水分、总灰分、酸不溶性灰分和水溶性浸出物含量。采用高效液相色谱法测定反枝苋药材中芦丁的含量,色谱柱为Agilent 5 TC-C1(82),流动相为甲醇-0.3%磷酸溶液(40∶60,V/V),检测波长为358 nm,流速为1.0 mL/min,柱温为30℃,进样量为10μL。结果:药材外观性状及显微结构特征与现有描述相符。TLC鉴别结果显示,7批药材供试品色谱中,在与各对照品(芦丁、缬氨酸、亮氨酸)色谱相应的位置上均显相同颜色的斑点。7批反枝苋药材中水分的含量为7.43%~8.72%、总灰分的含量为11.82%~13.78%、酸不溶性灰分的含量为0.15%~0.55%、水溶性浸出物的含量为17.27%~24.74%。芦丁检测质量浓度的线性范围为10~200μg/mL(R2=1.0000);精密度、稳定性(24 h)、重复性试验的RSD均小于2.0%(n=6);低、中、高质量浓度样品中芦丁的平均加样回收率分别为99.14%、97.98%、98.80%,RSD分别为0.97%、0.95%、0.96%(n=3)。7批反枝苋药材中芦丁的含量为0.314~1.102 mg/g。结论:本研究对反枝苋药材进行了性状观察、显微鉴别以及水分、总灰分、酸不溶性灰分和水溶性浸出物含量等检查项的考察,并建立了反枝苋药材中亮氨酸、缬氨酸和芦丁定性鉴别的TLC法以及芦丁含量测定的HPLC法,可为反枝苋药材质量标准的制订提供一定参考。

OBJECTIVE:To provide reference for the quality standard establishment of Amaranthus retroflexus.METHODS:Taking 7 batches of A.retroflexus medicinal materials as the research object,the appearance properties of the medicinal materials were investigated,and the microscopic characteristics of the medicinal powders were observed.TLC method was adopted to qualitatively identify rutin,valine and leucine in A.retroflexus medicinal materials.According to the relevant methods of the 2015 edition of Chinese Pharmacopoeia(partⅣ),water content,total ash content,acid-insoluble ash content and water-soluble extract content were determined.HPLC method was used to determine the content of rutin in the medicinal material of A.retroflexus.The determination was performed on Agilent 5 TC-C18(2)column with mobile phase consisted of methanol-0.3%phosphoric acid solution(40∶60,V/V),at the flow rate of 1.0 mL/min.The detection wavelength was set at 358 nm,and the column temperature was 30℃.The sample size was 10μL.RESULTS:The appearance and microstructure characteristics of the medicinal materials were consistent with the existing description.The identification results of TLC method showed that 7 batches of medicinal materials and each reference substance(rutin,valine,leucine)showed spots of the same color at the same position.The moisture content of 7 batches of A.retroflexus medicinal materials was 7.43%-8.72%,the total ash content was 11.82%-13.78%,the acid-insoluble ash content was 0.15%-0.55%,and the water soluble extract content was 17.27%-24.74%.The linear range of rutin was 10-200μg/mL(R 2=1.0000).RSDs of precision test,stability test(24 h)and repeatability test were all less than 2.0%(n=6).The average recovery rates of rutin were 99.14%,97.98%and 98.80%in low,medium and high concentration of samples,and RSDs were 0.97%,0.95%,0.96%(n=3).The contents of rutin in 7 batches of A.retrophylla were 0.314-1.102 mg/g.CONCLUSIONS:In this study,character observation,microscopic identification,moisture content,total ash content,acid-insoluble ash content and water-soluble extract content of A.retroflexus are investigated;TLC method was established for qualitative identification of leucine,valine and rutin in A.retroflexus,and the HPLC method was established for content determination of rutin.It provides reference for the quality standard establishment of A.retroflexus.