胡桃楸枝和根中5个成分含量测定方法的建立及其含量差异分析  

Establishment of Content Determination of 5 Components and Their Content Difference Analysis in the Branch and Root of Juglans mandshurica

作　　者：刘宏 宋奇[1] 王添敏[1] 张慧[1] 翟延君[1] 康廷国[1] LIU Hong;SONG Qi;WANG Tianmin;ZHANG Hui;ZHAI Yanjun;KANG Tingguo(School of Pharmacy,Liaoning University of TCM,Liaoning Dalian 116600,China)

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600

出　　处：《中国药房》2021年第8期933-939,共7页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81703658);辽宁省自然科学基金资助计划(No.2019-MS-223);沈阳市中青年科技创新人才支持计划项目(No.RC190083)。

摘　　要：目的:建立同时测定胡桃楸枝和根中没食子酸、逆没食子酸、1,6-二-O-没食子酰基-β-D-葡萄糖、1,2,6-三-O-没食子酰基-β-D-葡萄糖、1,2,3,6-四-O-没食子酰基-β-D-葡萄糖等5个成分含量的方法,并分析枝和根中上述5个成分的含量差异。方法:采用高效液相色谱法,以Agilent Poroshell 120 SB-C18为色谱柱,以水(含0.2%甲酸)-乙腈(含0.2%甲酸)为流动相进行梯度洗脱,流速为0.3 mL/min,柱温为30℃,检测波长为270 nm,进样量为5μL。利用独立样本t检验和偏最小二乘法判别分析对5个成分含量进行比较分析。结果:没食子酸、逆没食子酸、1,6-二-O-没食子酰基-β-D-葡萄糖、1,2,6-三-O-没食子酰基-β-D-葡萄糖、1,2,3,6-四-O-没食子酰基-β-D-葡萄糖检测质量浓度的线性范围分别为0.989~63.3、1.58~101、1.01~64.7、3.31~212、3.34~214μg/mL(r≥0.9973);精密度、重复性、稳定性(12 h)试验的RSD均小于3.2%,平均加样回收率分别为103.2%、99.1%、101.5%、102.9%、104.7%(RSD分别为4.85%、2.80%、1.31%、2.73%、1.28%)。在胡桃楸枝中,上述成分的平均含量分别为0.2965、0.6211、0.5625、3.1117、3.4513 mg/g;在根中,上述成分的平均含量分别为0.6734、2.7555、0.9640、2.9466、4.8364 mg/g;在枝、根中上述成分的平均总量分别为8.0432、12.1759 mg/g。根中没食子酸、逆没食子酸和1,6-二-O-没食子酰基-β-D-葡萄糖含量均显著高于枝中的含量(P<0.05或P<0.01),而枝和根中其余2个成分的含量及5个成分的总量比较差异均无统计学意义(P>0.05)。偏最小二乘法判别分析所建模型的累积解释度(R^(2)X、R^(2)Y)、累积预测度(Q^(2))分别为0.943、0.745、0.710;模型载荷图显示,逆没食子酸与原点距离最远,仅逆没食子酸含量的变量投影重要性值大于1。结论:成功建立了可同时测定胡桃楸枝和根中5个成分含量的方法。除1,2,6-三-O-没食子酰基-β-D-葡萄糖外,胡桃楸根中其余4个成分的含量及总含OBJECTIVE:To establish a method for simultaneous determination of 5 components in the branch and root of Juglans mandshurica as gallic acid,ellagic acid,1,6-di-O-galloyl-β-D-glucose,1,2,6-tri-O-galloyl-β-D-glucose and 1,2,3,6-tetra-O-galloyl-β-D-glucose,and to analyze the content difference of above 5 components between the branch and root samples.METHODS:HPLC method was adopted.The determination was performed on Agilent Poroshell 120 SB-C18 column with mobile phase consisted of water(containing 0.2%formic acid)-acetonitrile(containing 0.2%formic acid).A gradient elution was performed at a flow rate of 0.3 mL/min.The column temperature was 30℃and the detection wavelength was 270 nm.The sample size was 5μL.Independent samples t-test and partial least squares-discriminant analysis(PLS-DA)were applied for statistical analysis of 5 components.RESULTS:The linear range of gallic acid,ellagic acid,1,6-di-O-galloyl-β-D-glucose,1,2,6-tri-O-galloyl-β-D-glucose and 1,2,3,6-tetra-O-galloyl-β-D-glucose were 0.989-63.3,1.58-101,1.01-64.7,3.31-212,3.34-214μg/mL(r≥0.9973),respectively.RSDs of precision,reproducibility and stability tests(12 h)were all lower than 3.2%.The average recoveries of the 5 components were 103.2%(RSD=4.85%),99.1%(RSD=2.80%),101.5%(RSD=1.31%),102.9%(RSD=2.73%)and 104.7%(RSD=1.28%),respectively.The average contents of the above components in the branch of J.mandshurica were 0.2965,0.6211,0.5625,3.1117 and 3.4513 mg/g,respectively.The average contents of above components in the root were 0.6734,2.7555,0.9640,2.9466 and 4.8364 mg/g,respectively.The total contents of the 5 components in the branch and root of J.mandshurica were 8.0432 and 12.1759 mg/g,respectively.The contents of gallic acid,ellagic acid and 1,6-di-O-galloyl-β-D-glucose in roots were significantly higher than those in branches(P<0.05 or P<0.01).There were no significant differences in the contents of the other 2 components and the total contents of the 5 components in branches and roots(P>0.05).The cumulative interpretability(R 2

关 键 词：胡桃楸 枝 根 没食子酸 逆没食子酸 鞣质 含量测定 

分 类 号：R927.2[医药卫生—药学]