吴茱萸、穿心莲等中药材化学成分分离纯化方法研究

发布时间：2019-04-08 15:56

【摘要】：我国传统中药种类繁多,各种药材所含有的化学成分很是复杂,含量也相对较少。因而,从传统中药中分离纯化出有效成分对于临床医学的药理作用研究及含量的测定具有较为深远的影响。本文分别应用超临界流体色谱(supercritical fluid chromatography,SFC)和制备型高效液相色谱(preparative high performance liquid chromatography,PHPLC)对吴茱萸、穿心莲、木蝴蝶及知母中的化学成分进行分离与纯化,所得到的化合物的纯度经高效液相色谱(high performance liquid chromatography,HPLC)进行了检测,并采用质谱(MS)、紫外光谱(UV)和核磁共振波谱(NMR)对所得到的化合物的结构进行了鉴定。1.SFC分离纯化吴茱萸中两种生物碱类成分研究使用超临界二氧化碳(SC-CO_2)作为SFC流动相的主体,添加一定量的改性剂(如:甲醇、乙醇、异丙醇、乙腈等),建立了从吴茱萸中分离纯化吴茱萸碱与吴茱萸次碱等成分的SFC方法。在一定研究范围内,通过考察色谱柱的类型、改性剂种类及含量、流速、压力和温度等条件对分离效果的影响,最终确立了从吴茱萸中分离纯化吴茱萸碱与吴茱萸次碱的最适宜的色谱条件。从吴茱萸中分离纯化得到的化合物的纯度经HPLC检测分别为99.5%和99.8%,经UV和NMR鉴定得到吴茱萸碱与吴茱萸次碱的化学结构。通过实验数据,拟合得到van’t Hoff曲线,进一步探讨了SFC分离过程的热力学变化规律。将吴茱萸碱与吴茱萸次碱从流动相到固定相分配过程中的焓变差值与熵变差值进行比较,结果表明:在该条件下,从吴茱萸中分离纯化吴茱萸碱与吴茱萸次碱的SFC过程为焓控制的过程。2.SFC分离纯化穿心莲中两种二萜内酯类成分研究使用SC-CO_2作为SFC流动相的主体,添加一定量的改性剂(如:甲醇、乙醇、异丙醇、乙腈等),建立了从穿心莲中分离纯化穿心莲内酯与脱水穿心莲内酯等化学成分的SFC方法。在一定研究范围内,通过考察色谱柱的类型、改性剂种类及含量、流速、压力、温度等条件对分离效果的影响,最终确立了从穿心莲中分离纯化穿心莲内酯和脱水穿心莲内酯的最适宜的色谱条件。从穿心莲中分离纯化得到的化合物的纯度经HPLC检测分别为99.1%和98.5%,经UV和NMR鉴定得到穿心莲内酯与脱水穿心莲内酯的化学结构。通过实验数据,拟合得到van’t Hoff曲线,进一步探讨了SFC分离过程的热力学变化规律。将穿心莲中的穿心莲内酯与脱水穿心莲内酯从流动相到固定相分配过程中的焓变差值与熵变差值进行比较,结果表明:在该条件下,从穿心莲中分离纯化穿心莲内酯与脱水穿心莲内酯的SFC过程为焓控制的过程。3.PHPLC分离纯化木蝴蝶中的两种黄酮类成分实验中以甲醇-水为流动相,建立了从木蝴蝶中分离纯化黄芩素-7-O-葡萄糖苷和黄芩素-7-O-双葡萄糖苷的PHPLC方法。采用C18柱(250 mm×25.4 mm I.D.,10μm),以甲醇-水(45:55,V/V)作为流动相进行等度洗脱,上样量为20 mg,流速为30 m L/min,检测波长设定为280 nm。从木蝴蝶中分离纯化所得到的两种化合物的纯度经HPLC检测分别为98.1%和98.9%;然后再采用MS、UV、NMR鉴定,得到黄芩素-7-O-葡萄糖苷和黄芩素-7-O-双葡萄糖苷的化学结构。4.PHPLC分离纯化知母中的两种黄酮类成分实验中以甲醇-水为流动相,建立了从知母中分离纯化的芒果苷和新芒果苷的PHPLC方法。采用C18柱(250 mm×25.4 mm I.D.,10μm),流动相为甲醇-水(35:65,V/V)等度洗脱,上样量为27.6 mg,流速为35 m L/min,检测波长设定为254 nm。从知母中分离得到的两种单体成分,经HPLC检测,其纯度分别为98.4%和99.1%;其化学结构经MS、UV、NMR鉴定为芒果苷和新芒果苷。
[Abstract]:The traditional Chinese medicine has a wide variety of traditional Chinese medicines, and the chemical components contained in the various medicinal materials are very complex and the content is relatively small. Therefore, the separation and purification of the effective components from the traditional Chinese medicine has a far-reaching influence on the pharmacological action of the clinical medicine and the determination of the content. The chemical constituents of Wu's, Andrographis and Anemarrhena are isolated and purified by supercritical fluid chromatography (SFC) and preparative high performance liquid chromatography (PHPLC), respectively. the purity of the obtained compound is detected by high performance liquid chromatography (hplc) and mass spectrometry (ms) is used, The structure of the obtained compound was identified by ultraviolet (UV) and nuclear magnetic resonance spectroscopy (NMR). In this paper, the SFC method for separating and purifying the components such as Wu's base and the base of Wu's tribasic was established. In this study, the most suitable chromatographic conditions for the separation and purification of Wu's base and the base of Wu's tribasic were established by examining the influence of the type of the column, the type of the modifier, the content, the flow rate, the pressure and the temperature on the separation effect. The purity of the obtained compound was 99.5% and 99.8% by HPLC, and the chemical structure of Wu's base and Wu's tribasic was obtained by UV and NMR. According to the experimental data, the van't Hoff curve is obtained, and the thermodynamic rule of the SFC separation process is further discussed. In this paper, the difference of the difference between the temperature and the entropy of the alkali from the mobile phase to the fixed phase is compared with the difference of the entropy variation. The results show that under this condition, In this paper, the SFC process of the purification of the two kinds of diolactone in Andrographis paniculata was studied by using SC-CO _ 2 as the main body of the SFC mobile phase, and a certain amount of modifier (such as methanol, ethanol, isopropanol, acetonitrile, etc.) was added. A method for separating and purifying the chemical constituents of Andrographolide and dehydroandrographolide from Andrographis paniculata is established. The most suitable chromatographic conditions for separating and purifying andrographolide and dehydroandrographolide from Andrographis paniculata were established by examining the influence of the type of the column, the type of the modifier, the content, the flow rate, the pressure and the temperature on the separation effect. The purity of the compound obtained from the separation and purification of the Andrographis paniculata is 99.1% and 98.5%, respectively, and the chemical structure of the andrographolide and the dehydrated andrographolide is obtained by UV and NMR. According to the experimental data, the van't Hoff curve is obtained, and the thermodynamic rule of the SFC separation process is further discussed. comparing the pressure difference between the andrographolide and the dehydrated andrographolide from the flowing phase to the fixed phase distribution process, and the result shows that under the condition, the process of separating and purifying the SFC process of the andrographolide and the dehydrated andrographolide from the Andrographis paniculata is the process of the control of the water,3. the two kinds of flavonoids in the purified wood butterfly are separated and purified by the PHPLC, and the methanol-water is used as the mobile phase, A PHPLC method for separating and purifying the berberine-7-O-glucose and the berberine-7-O-digluconate from a butterfly was established. A C18 column (250 mm, 25.4 mm I. D.,10. mu.m) was used as the mobile phase for isocratic elution with methanol-water (45:55, V/ V), with a loading amount of 20 mg, a flow rate of 30 m L/ min, and a detection wavelength of 280 nm. The purity of the two compounds obtained from the separation and purification of the wood butterfly is 98.1% and 98.9%, respectively, and then the MS, the UV and the NMR are used for identification, The chemical structure of the berberine-7-O-glucomin and the berberine-7-O-digluconate was obtained.4. The PHPLC was used to separate and purify the two kinds of flavonoids in anemarrhena, and a PHPLC method for separating and purifying the mango and the new mango juice from anemarrhena was established. A C18 column (250 mm, 25.4 mm I. D.,10. mu.m) was used, and the mobile phase was methanol-water (35:65, V/ V) isocratic elution, the loading amount was 27.6 mg, the flow rate was 35 m L/ min, and the detection wavelength was set to 254 nm. The purity of the two monomer components isolated from Rhizoma Anemarrhenae was 98.4% and 99.1%, respectively. The chemical structure of the two monomer components was identified as mango juice and new mango juice by MS, UV and NMR.
【学位授予单位】：聊城大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284.1

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