不同采收期赤芍中6种有效成分的含量测定及其变化规律研究
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篇名: 不同采收期赤芍中6种有效成分的含量测定及其变化规律研究
TITLE: Study on Content Determination of 6 Active Ingredients in Paeonia lactiflora during Different Harvesting Periods and Their Variation
摘要: 目的:建立同时测定不同采收期赤芍中6种有效成分含量的方法,并探讨其变化规律,以确定最佳采收期。方法:采用高效液相色谱法测定样品中没食子酸、儿茶素、芍药内酯苷、芍药苷、苯甲酸、苯甲酰芍药苷的含量并进行主成分分析。色谱柱为ThermoC18,流动相为乙腈-0.1%磷酸水溶液(梯度洗脱),流速为1.0mL/min,检测波长为230nm,柱温为35℃,进样量为10µL。结果:上述6种成分的质量浓度线性范围分别为0.0132~0.25mg/mL(r=0.9992)、0.0132~0.25mg/mL(r=0.9999)、0.0268~0.51mg/mL(r=0.9997)、0.42~8.01mg/mL(r=0.9992)、0.016~0.31mg/mL(r=0.9994)、0.02~0.38mg/mL(r=0.9998);定量限分别为0.0093、0.0085、0.0163、0.0217、0.0113、0.0174mg/mL,检测限分别为0.0033、0.0027、0.0054、0.0073、0.0038、0.0059mg/mL;精密度、稳定性、重复性试验的RSD均小于2%;加样回收率为96.01%~99.43%(RSD=1.23%,n=9)、97.95%~100.45%(RSD=0.79%,n=9)、97.98%~100.11%(RSD=0.68%,n=9)、98.83%~100.09%(RSD=0.65%,n=9)、98.58%~100.95%(RSD=1.35%,n=9)、96.28%~103.26%(RSD=1.76%,n=9)。赤芍药材(根)中上述6种成分的含量分别为0.016%~0.057%、0~0.067%、0.207%~0.640%、2.350%~5.887%、0.030%~0.245%、0.054%~0.381%。5月30日采收的赤芍药材折干率最低(约33%),9月15日折干率最高(约49%)。7-10月采收的赤芍叶中没食子酸、芍药内酯苷的含量均高于根部。主成分分析结果显示,前2个主成分的方差贡献率分别为71.845%、18.170%,累积方差贡献率为90.015%;综合得分较高的月份为5-6月、9-10月。结论:所建含量测定方法简便、准确,重复性、精密度较好,可用于测定不同采收期赤芍药材中6种有效成分的含量测定;赤芍药材的最佳采收期为9月30日-10月15日,叶可在7月15日前后采收。
ABSTRACT: OBJECTIVE:To establish the method for content determination of 6 active ingredients in Paeonia lactiflora during different harvest periods ,and to investigate its variation rules so as to determine the optimal harvesting period. METHODS :HPLC method was adopted to determine the contents of gallic acid , catechin, alibiflorin, paeoniflorin, benzoic acid and benzoylpaeoniflorin,and principal component analysis was conducted . The determination was performed on Thermo C 18 column with mobile phase consisted of acetonitrile- 0.1% phosphoric acid aqueous solution (gradient elution )at the flow rate of 1.0 mL/min. The detection wavelength was 230 nm,and column temperature was 35 ℃. The sample size was 10 µL. RESULTS :The linear range of the above 6 ingredients were 0.013 2-0.25 mg/mL(r=0.999 2),0.013 2-0.25 mg/mL(r=0.999 9),0.026 8-0.51 mg/mL(r=0.999 7), 0.42-8.01 mg/mL(r=0.999 2),0.016-0.31 mg/mL(r=0.999 4),0.02-0.38 mg/mL(r=0.999 8),respectively. The limits of quantification were 0.009 3,0.008 5,0.016 3,0.021 7,0.011 3,0.017 4 mg/mL,and the limits of detection were 0.003 3,0.002 7, 0.005 4,0.007 3,0.003 8,0.005 9 mg/mL. RSDs of precision ,stability and repeatability tests were less than 2%. The recoveries were 96.01%-99.43%(RSD=1.23%,n=9),97.95%-100.45%(RSD=0.79%,n=9),97.98%-100.11%(RSD=0.68%,n= 9),98.83% -100.09% (RSD=0.65% ,n=9),98.58% - 100.95%(RSD=1.35%,n=9),96.28%-103.26%(RSD= 1.76%,n=9). The contents of above 6 ingredients in Radix Paeoniae Rubra (roots) were 0.016% -0.057% ,0-0.067% , 0.207%-0.640%,2.350%-5.887%,0.030%- 0.245%,0.054%- 0.381%,respectively. On May 30th,the drying rate of Radix 0451-87266873。E-mail:wangzhen_yue@163.com Paeoniae Rubra was the lowest (about 33%),and on Sept. 15th,the drying rate was the highest (about 49%). The contents of gallic acid and paeoniflorin in the leaves of P. lactiflora were higher than the root during Jul.-Oct. Results of principal component analysis showed that the variance contribution rates of the first two principal components were 71.845% and 18.170%,respectively;cumulative variance contribution rate was 90.015%. The months with higher comprehensive scores were May to Jun. and Sept. to Oct. CONCLUSIONS :Established method is simple , accurate,reproducible and precise. It can be used to determine the contents of 6 active ingredients in Radix Paeoniae Rubra during different harvest periods. Sept. 30th to Oct. 15th is the optimum harvesting periods for Radix Paeoniae Rubra ,and leaves can be harvested around Jul. 15th.
期刊: 2020年第31卷第04期
作者: 付士朋,沈宏伟,王谦博,李俊萍,王聪,郭盛磊,王振月
AUTHORS: FU Shipeng ,SHEN Hongwei ,WANG Qianbo ,LI Junping ,WANG Cong,GUO Shenglei ,WANG Zhenyue
关键字: 赤芍;根;叶;采收期;高效液相色谱法;含量测定;变化规律;主成分分析
KEYWORDS: Paeonia lactiflora ;Roots;Leaves;Harvesting period ;HPLC;Content determination ;Variation rule ;Principal
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