固相萃

发布时间：2019-02-09 20:30

【摘要】：中药成分复杂,将有效成分富集并测定其含量是研究开发中药的重要环节。qNMR方法无需被测物的高纯标准品,只需常用的几种内标和氘代试剂就可完成定量分析,但存在检测灵敏度和干扰问题。SPE作为一种方便有效的样品前处理技术,具有浓缩(富集)和样品净化功能,操作简便、回收率高。本文将SPE与qNMR结合,利用SPE的富集作用显著拓展了 qNMR用于低含量复杂样品的定量分析范围。本文以板蓝根饮片、乳增宁胶囊和双黄连胶囊为样品,分别建立了 SPE-qNMR测定三种中药中有效成分含量的方法,对所建立的方法进行了验证,并在无需待测物高纯标准品的情况下分别测定了板蓝根饮片中表告依春、乳增宁胶囊中淫羊藿苷和双黄连胶囊中绿原酸的含量。1.建立了 SPE-qNMR测定板蓝根饮片中的有效成分表告依春含量的方法。样品用水两次60℃超声提取,以Poly-Sery MCX(混合型强阳离子交换)型SPE柱对提取液富集浓缩后,用qNMR测定表告依春的含量。考察了超声时间、SPE样品前处理条件的选择以及定量核磁共振技术的实验条件对定量结果的影响。选择氘代二甲基亚砜为氘代溶剂,2,3,5-三碘苯甲酸作为内标,并且用基准试剂邻苯二甲酸氢钾对其进行标定。选择脉冲宽度P1=14.1μs,延迟时间d1=5s,扫描次数NS=256为qNMR定量表告依春的实验条件。表告依春的定量峰为δ5.365～5.399 (H-7b, d,1H)。结果显示,所建方法的日内精密度的RSD为0.47%,日间精密度RSD为0.80%,表告依春与三碘苯甲酸峰面积比与质量比的零截距标准曲线线性相关系数为0.9991,且斜率与理论值相符。该法测定表告依春的LOD为0.05 mg/g; LOQ为0.19mg/g。包括样品预处理过程的表告依春的回收率为97.4%～101.7%。实际测定板蓝根饮片样品中的表告依春的含量为0.19 ～1.26 mg/g。2.建立了 SPE-qNMR测定乳增宁胶囊中有效成分淫羊藿苷含量的方法。样品用10%乙醇室温超声提取,以HC-C18型SPE柱对提取液进行浓缩除杂后,用qNMR测定淫羊藿苷的含量。考察了超声时间、SPE样品前处理条件以及定量核磁共振实验条件对定量结果的影响。选择氘代二甲基亚砜为氘代溶剂,2,3,5-三碘苯甲酸作为内标,并且用基准试剂邻苯二甲酸氢钾对其进行标定。选择脉冲宽度P1=14.1μs,延迟时间d1=1s,扫描次数NS=256为qNMR定量淫羊藿苷的实验条件。淫羊藿苷的定量峰为δ7.890～7.909(2',6'-H,d,2H)。结果显示,所建方法的日内精密度的RSD为0.43%,日间精密度RSD为0.75%,淫羊藿苷与三碘苯甲酸峰面积比与质量比的零截距标准曲线线性相关系数为0.9999,且斜率与理论值相符。该法测定淫羊藿苷的LOD为0.122mg/g; LOQ为0.368mg/g。包括样品预处理过程的淫羊藿苷的回收率为99.9%～102.9%。实际测定乳增宁胶囊中的淫羊藿苷的含量为3.947～4.392mg/g。3.建立了 SPE-qNMR测定双黄连胶囊中有效成分绿原酸含量的方法。样品用纯水室温超声提取,以HC-C18型SPE柱对提取液进行浓缩除杂后,用qNMR测定绿原酸的含量。考察了超声时间、SPE样品前处理条件以及定量核磁共振实验条件对定量结果的影响。选择氘代二甲基亚砜为氘代溶剂,对苯二甲醛作为内标,并且用基准试剂邻苯二甲酸氢钾对其进行标定。选择脉冲宽度P1=14.1μs,延迟时间d1=1s,扫描次数NS=512为qNMR定量绿原酸的实验条件。绿原酸的定量峰为δ6.149-6.182(H-8',d,1H)。结果显示,所建方法的日内精密度的RSD为1.2%,日间精密度RSD为1.5 %,绿原酸与对苯二甲醛峰面积比与质量比的零截距标准曲线线性相关系数为0.9999,且斜率与理论值相符。该法测定绿原酸的LOD为0.0017mg/g; LOQ为0.0791mg/g。包括样品经SPE预处理过程的绿原酸的回收率为101.3%～104.4%。实际测定双黄连胶囊中的绿原酸的含量为9.68～10.35 mg/g。
[Abstract]:The composition of the traditional Chinese medicine is complex, the effective components are enriched and the content of the traditional Chinese medicine is determined to be an important step in the research and development of the traditional Chinese medicine. The qNMR method does not need the high-purity standard of the object to be measured, and the quantitative analysis can be carried out only by using the common internal standard and the generation reagent, but the detection sensitivity and the interference problem exist. As a convenient and effective sample pretreatment technique, the SPE has the functions of concentration (enrichment) and sample purification, simple and convenient operation and high recovery rate. In this paper, the SPE is combined with qNMR, and the concentration of SPE is used to greatly expand the quantitative range of quantitative analysis of low-content complex samples by qNMR. In this paper, the method of SPE-qNMR to determine the content of effective components in the three traditional Chinese medicines was established by using the method of SPE-qNMR to determine the content of the effective components in the three traditional Chinese medicines. in that method, the content of the chlorogenic acid in the radix isatidis decoction pieces and the content of the chlorogenic acid in the two coptis capsules are respectively measured in the radix isatidis decoction pieces without the high pure standard of the object to be measured. The method of SPE-qNMR to determine the content of Radix Isatidis in Radix Isatidis decoction pieces was established. The sample was extracted with water twice with water for 60 鈩，

本文编号：2419364