發(fā)布時間：2019-02-09 20:30

【摘要】：中藥成分復(fù)雜,將有效成分富集并測定其含量是研究開發(fā)中藥的重要環(huán)節(jié)。qNMR方法無需被測物的高純標(biāo)準(zhǔn)品,只需常用的幾種內(nèi)標(biāo)和氘代試劑就可完成定量分析,但存在檢測靈敏度和干擾問題。SPE作為一種方便有效的樣品前處理技術(shù),具有濃縮(富集)和樣品凈化功能,操作簡便、回收率高。本文將SPE與qNMR結(jié)合,利用SPE的富集作用顯著拓展了 qNMR用于低含量復(fù)雜樣品的定量分析范圍。本文以板藍根飲片、乳增寧膠囊和雙黃連膠囊為樣品,分別建立了 SPE-qNMR測定三種中藥中有效成分含量的方法,對所建立的方法進行了驗證,并在無需待測物高純標(biāo)準(zhǔn)品的情況下分別測定了板藍根飲片中表告依春、乳增寧膠囊中淫羊藿苷和雙黃連膠囊中綠原酸的含量。1.建立了 SPE-qNMR測定板藍根飲片中的有效成分表告依春含量的方法。樣品用水兩次60℃超聲提取,以Poly-Sery MCX(混合型強陽離子交換)型SPE柱對提取液富集濃縮后,用qNMR測定表告依春的含量。考察了超聲時間、SPE樣品前處理條件的選擇以及定量核磁共振技術(shù)的實驗條件對定量結(jié)果的影響。選擇氘代二甲基亞砜為氘代溶劑,2,3,5-三碘苯甲酸作為內(nèi)標(biāo),并且用基準(zhǔn)試劑鄰苯二甲酸氫鉀對其進行標(biāo)定。選擇脈沖寬度P1=14.1μs,延遲時間d1=5s,掃描次數(shù)NS=256為qNMR定量表告依春的實驗條件。表告依春的定量峰為δ5.365～5.399 (H-7b, d,1H)。結(jié)果顯示,所建方法的日內(nèi)精密度的RSD為0.47%,日間精密度RSD為0.80%,表告依春與三碘苯甲酸峰面積比與質(zhì)量比的零截距標(biāo)準(zhǔn)曲線線性相關(guān)系數(shù)為0.9991,且斜率與理論值相符。該法測定表告依春的LOD為0.05 mg/g; LOQ為0.19mg/g。包括樣品預(yù)處理過程的表告依春的回收率為97.4%～101.7%。實際測定板藍根飲片樣品中的表告依春的含量為0.19 ～1.26 mg/g。2.建立了 SPE-qNMR測定乳增寧膠囊中有效成分淫羊藿苷含量的方法。樣品用10%乙醇室溫超聲提取,以HC-C18型SPE柱對提取液進行濃縮除雜后,用qNMR測定淫羊藿苷的含量�？疾炝顺�聲時間、SPE樣品前處理條件以及定量核磁共振實驗條件對定量結(jié)果的影響。選擇氘代二甲基亞砜為氘代溶劑,2,3,5-三碘苯甲酸作為內(nèi)標(biāo),并且用基準(zhǔn)試劑鄰苯二甲酸氫鉀對其進行標(biāo)定。選擇脈沖寬度P1=14.1μs,延遲時間d1=1s,掃描次數(shù)NS=256為qNMR定量淫羊藿苷的實驗條件。淫羊藿苷的定量峰為δ7.890～7.909(2',6'-H,d,2H)。結(jié)果顯示,所建方法的日內(nèi)精密度的RSD為0.43%,日間精密度RSD為0.75%,淫羊藿苷與三碘苯甲酸峰面積比與質(zhì)量比的零截距標(biāo)準(zhǔn)曲線線性相關(guān)系數(shù)為0.9999,且斜率與理論值相符。該法測定淫羊藿苷的LOD為0.122mg/g; LOQ為0.368mg/g。包括樣品預(yù)處理過程的淫羊藿苷的回收率為99.9%～102.9%。實際測定乳增寧膠囊中的淫羊藿苷的含量為3.947～4.392mg/g。3.建立了 SPE-qNMR測定雙黃連膠囊中有效成分綠原酸含量的方法。樣品用純水室溫超聲提取,以HC-C18型SPE柱對提取液進行濃縮除雜后,用qNMR測定綠原酸的含量。考察了超聲時間、SPE樣品前處理條件以及定量核磁共振實驗條件對定量結(jié)果的影響。選擇氘代二甲基亞砜為氘代溶劑,對苯二甲醛作為內(nèi)標(biāo),并且用基準(zhǔn)試劑鄰苯二甲酸氫鉀對其進行標(biāo)定。選擇脈沖寬度P1=14.1μs,延遲時間d1=1s,掃描次數(shù)NS=512為qNMR定量綠原酸的實驗條件。綠原酸的定量峰為δ6.149-6.182(H-8',d,1H)。結(jié)果顯示,所建方法的日內(nèi)精密度的RSD為1.2%,日間精密度RSD為1.5 %,綠原酸與對苯二甲醛峰面積比與質(zhì)量比的零截距標(biāo)準(zhǔn)曲線線性相關(guān)系數(shù)為0.9999,且斜率與理論值相符。該法測定綠原酸的LOD為0.0017mg/g; LOQ為0.0791mg/g。包括樣品經(jīng)SPE預(yù)處理過程的綠原酸的回收率為101.3%～104.4%。實際測定雙黃連膠囊中的綠原酸的含量為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，

本文編號：2419364