摘要由于中药成分十分复杂，所以研究开发中药的重要环节就是将有效成分富集并测定其含量。定量核磁共振方法不需要被测物的高纯度标准品，要完成定量分析只需要几种常用的内标与氘代试剂，只是存在干扰问题与检测灵敏度的问题。固相萃取是一种非常方便有效的样品前处理的技术，它具有样品净化与浓缩(富集)的功能，回收率高、操作简单。本文将固相萃取和定量核磁共振相结合，利用固相萃取的富集作用显著的拓展了定量核磁共振用于低含量复杂样品分析时的定量分析范围。82291

    乳增宁胶囊作为本文的样品，建立了固相萃取-定量核磁共振测定乳增宁胶囊中的有效成分含量的方法，同时验证了所建立的方法，并且在不需要待测物高纯标准品的条件下测定了乳增宁胶囊中ICA（淫羊藿苷）的含量。

    建立了固相萃取-定量核磁共振测定乳增宁胶囊中有效成分ICA含量的方法。乳增宁胶囊先用10%乙醇室温超声提取，再用HC-C18型固相萃取柱对提取液进行浓缩除杂后，最后用定量核磁共振测定ICA的含量。整体对超声的时间、固相萃取样品前处理的条件和定量核磁共振实验条件对定量结果的影响进行了考察，氘代溶剂选择为氘代二甲基亚砜，内标为2,3,5-三碘苯甲酸，并且用选择邻苯二甲酸氢钾为基准试剂对其进行标定，脉冲宽度的选择为P1=14。1µs，延迟时间为d1=1s，扫描次数为NS=256，这些都是定量核磁共振定量ICA的实验条件。δ7。890~7。909 (2’,6’-H,d,2H)为ICA的定量峰。根据结果可知，所建立的方法的日内精密度RSD为0。43%，日间精密度RSD为0。75%，0。9999为ICA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，并且斜率与理论值相符。这种方法测定ICA的LOD为0。122mg/g；LOQ为0。368mg/g。99。9%~102。9%为样品预处理过程的ICA的回收率。实际测定的乳增宁胶囊中的ICA的含量为3。947~4。392mg/g。

毕业论文关键词：固相萃取；定量核磁共振技术；淫羊藿苷

Determination of  Icariin in Ruzengning capsule by Solid Phase Extraction - Quantitative Nuclear Magnetic Resonance SPE ctroscopy

Abstract

The determination of the active ingredients is a key point to the development of traditional Chinese medicine。 Without the high purity standard compound of the test substance, quantitative nuclear magnetic resonance (qNMR) can analyze sample accurately and fastly via using several common internal standards and deuterated reagents。 But the detection sensitivity and interference are its shortcomings。 Solid phase extraction ( SPE), as an effective sample pretreatment technology, with sample enrichment and purification function, is easy to operate and have high recovery rate。 Combined SPE with qNMR will extend qNMR application in the low content components in complex sample。

In the thesis, the  SPE-qNMR method for the determination of active ingredients ICA in Ruzengning capsule was established。 And the established method was validated。 Without the high purity standard compounds, we determination of  ICA in Ruzengning by  SPE-qNMR 。The SPE is suggested as extender for qNMR application in the low content components in complex sample。

    The SPE-qNMR method for the determination of ICA in Ruzengning capsule was established。 The ultrasonic extraction method was used for fully extracting epigoitrin in sample using 10% a LC ohol as solvent at room temperature, and then the extracting solution was enriched and cleaned by HC-C18 SPE cartridge。 The effect of ultrasonic extraction, SPE sample pretreatment and qNMR experimental conditions were investigated。 The qNMR experiment conditions were selected using deuterated D MS O as solvent, calibrated 2,3,5-triiodobenzoate as internal standard, and P1(pulse width) = 14。4μs, d1(pulse delay time) = 1s, NS(number of scan) = 256。 The 1H-NMR peaks of 7。890~7。909ppm (2’,6’-H,d,2H) of icariin was chosen as the quantitative peaks。 Method validation was performed including precision (the intra-day precision RSD was 0。43% , and the inter-day precision was 0。75% ); linearity (correlation coefficient r > 0。9999); LOD (0。122 mg/g) and LOQ (0。368mg/g)。 The recovery of the SPE- qNMR was in 99。9%~102。9%。 The result shows the method stable, accurate and reliabile。 The determination of  ICA in a real Ruzengning was in 3。947~4。392mg/g。