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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
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| 篇名： | 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 |
| TITLE： | Establishmend of Content Determination of 5 Components and Their Content Difference Analysis in the Branch and Root of Juglans man dshurica |
| 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AgilentPoroshell120SB-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0.3mL/min，柱温为30℃，检测波长为270nm，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（12h）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、3.4513mg/g；在根中，上述成分的平均含量分别为0.6734、2.7555、0.9640、2.9466、4.8364mg/g；在枝、根中上述成分的平均总量分别为8.0432、12.1759mg/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（R2X、R2Y）、累积预测度（Q2）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量均高于胡桃楸枝，逆没食子酸是区分两类样品的主要差异成分。 |
| ABSTRACT： | OBJECTIVE：To esta blish a method for si multaneous 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 5 μL. 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 0.989-63.3，1.58-101，1.01-64.7，3.31-212，3.34-214 μg/mL （r≥0.997 3），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 0.296 5，0.621 1，0.562 5，3.111 7 and 3.451 3 mg/g，respectively. The average contents of above components in the root were 0.673 4，2.755 5，0.964 0，2.946 6 and 4.836 4 mg/g，respectively. The total contents of the 5 components in the branch and roo t of J. mandshurica were 8.043 2 and 12.175 9 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 2X，R 2Y） and cumulative predictability （Q 2） of the model established by PLS-DA were 0.943，0.745，and 0.710 respectively. The model load diagram showed that the distance between the ellagic acid and the origin was the farthest ，and only variable projection importance of the content of the ellagic acid was greater than 1. CONCLUSIONS：The established method can be used for the content determination of 5 components in the branch and root of J. mandshurica . Except for 1，2，6-tri-O-galloyl-β-D-glucose，the contents of other 4 components and total contents of the 5 components in the root of J. mandshurica are higher than those of the branch. Ellagic acid is selected as the potential marker for discriminating the branch and root samples. |
| 期刊： | 2021年第32卷第08期 |
| 作者： | 刘宏，宋奇，王添敏，张慧，翟延君，康廷国 |
| AUTHORS： | LIU Hong，SONG Qi，WANG Tianmin ，ZHANG Hui，ZHAI Yanjun ，KANG Tingguo |
| 关键字： | 胡桃楸；枝；根；没食子酸；逆没食子酸；鞣质；含量测定 |
| KEYWORDS： | Juglans mandshurica ；Branch；Root；Gallic acid ；Ellagic acid ；Tannins；Content determination |
| 阅读数： | 163 次 |
| 本月下载数： | 4 次 |
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