【摘要】：虎杖(Polygonum cuspidatum Sieb.et Zucc.)是我國的傳統(tǒng)中藥材之一,2000年版的《中國藥典》中被收錄于第一部�；⒄戎械纳锘钚猿煞钟熊晤惢衔锖忘S酮類化合物等,現(xiàn)代藥理學(xué)研究證明,芪類成分中白藜蘆醇(Resveratrol)的生理活性主要表現(xiàn)在抗氧化、抗自由基、抗衰老、抗癌、抗心血管疾病等方面;黃酮類化合物(Flavonoids)具有鎮(zhèn)痛、抗病毒、降血脂、降血糖、抗氧化等多種醫(yī)用功效。目前,從虎杖中分離純化其中有效成分的研究越來越受到人們的關(guān)注。本論文研究了虎杖中白藜蘆醇和黃酮類化合物的分離純化,主要取得以下結(jié)果:一、采用酶解法提取虎杖中的白藜蘆醇,通過單因素和正交設(shè)計(jì)實(shí)驗(yàn)優(yōu)化得到白藜蘆醇最優(yōu)提取工藝條件如下:提取溫度為45℃,提取時間為5.0 h,酶用量0.50%(m/m),提取液p H為6.5,白藜蘆醇提取率為75.66%,含量為3.80%,較原料藥中的含量(0.4%)提高了約10倍。二、考察了32種MAR及其混合床(MARs)對虎杖粗提物中白藜蘆醇的純化效果,結(jié)果表明,MARs LZ-94+LZ-78+LZ-76對白藜蘆醇的純化效果最好,優(yōu)化得出最佳吸附條件為:吸附液濃度為0.25mg/m L、吸附溫度45℃、吸附液體積60.0m L、吸附液p H=3.0、吸附時間9.0 h;最佳解吸條件為:解吸溫度55℃、解吸液乙醇體積分?jǐn)?shù)70%,解吸時間2.5h,解吸液p H=4.0。得出MARs對白藜蘆醇的吸附率97.96%,解吸率80.55%,最終白藜蘆醇回收率達(dá)到78.60%,純度達(dá)到19.32%,是白藜蘆醇粗提物的5.08倍。MARs對白藜蘆醇的吸附動力學(xué)研究表明MARs對白藜蘆醇的的吸附過程符合準(zhǔn)二級動力學(xué)模型,大孔區(qū)和中孔區(qū)的吸附過程均符合Intra-particle-diffusion模型,預(yù)示著吸附過程由粒內(nèi)擴(kuò)散過程控制,微孔區(qū)中的吸附過程由Homogeneous particle-diffusion模型控制;Freundlich模型更適合描述MARs對白藜蘆醇的熱力學(xué)吸附行為,說明MARs對虎杖中白藜蘆醇的吸附行為為多分子層物理吸附。三、由于在前部分對虎杖中的白藜蘆醇分離純化后,純度較低,所以本部分采用虎杖粗提物預(yù)處理法輔助MARs對白藜蘆醇進(jìn)行分離純化。首先研究了萃取方法和工藝,通過比較不同有機(jī)溶劑及混合溶劑對白藜蘆醇的萃取效果,得出以有機(jī)溶劑E/D=3/1為萃取劑時對白藜蘆醇萃取效果較好,萃取水相與有機(jī)相最佳體積比Vw/Vo=2/3、萃取時間間隔為60min、萃取5次最為適宜。該條件下,白藜蘆醇萃取率為88.81%,純度為66.50%。該萃取方法能夠在保證白藜蘆醇苷被充分水解成白藜蘆醇的前提下,保證白藜蘆醇最少量的分解損失,且由于E和D均是國際化學(xué)界公認(rèn)的綠色溶劑,環(huán)境友好。之后,將萃取后所得樣品分別采用硅膠柱層析技術(shù)和MARs技術(shù)進(jìn)行分離純化,比較了兩種分離純化方法,發(fā)現(xiàn)經(jīng)硅膠柱層析法分離純化后白藜蘆醇的回收率為61.60%,純度為73.70%,經(jīng)MARs技術(shù)純化白藜蘆醇后白藜蘆醇的回收率為81.10%,純度為87.12%。因此在純化相同質(zhì)量白藜蘆醇的前提下,MARs純化技術(shù)優(yōu)于硅膠柱層析法。四、采用超聲輔助法提取虎杖中黃酮類化合物,通過單因素實(shí)驗(yàn)和中心組合設(shè)計(jì)以及響應(yīng)面法優(yōu)化黃酮類化合物的提取工藝,得出最優(yōu)提取條件如下:提取液乙醇體積分?jǐn)?shù)為60%,提取溫度45℃、超聲提取時間34 min、超聲功率80W、固液比1/20(g/m L)、最佳提取次數(shù)為5次,該條件下虎杖中黃酮類化合物的提取率達(dá)到94.50%,其初始提取物的純度為31.00%。五、考察32種MAR及MARs對虎杖中黃酮類化合物的分離純化效果,結(jié)果得出單一MAR LZ-72對虎杖中黃酮類化合物的分離純化效果最佳,最佳吸附條件為:吸附原液濃度為2.50mg/m L,吸附溫度30℃,吸附液為10%的乙醇溶液、吸附液體積60.0 m L、吸附時間20 h;最佳解吸條件為:解吸溫度55℃、解吸液體積為110 m L,解吸液乙醇體積分?jǐn)?shù)為90%,解吸附時間為20 h,最終產(chǎn)物純度達(dá)到82.50%,回收率達(dá)到89.70%。LZ-72對黃酮類化合物的吸附動力學(xué)研究表明該吸附過程符合準(zhǔn)二級動力學(xué)模型,大孔區(qū)和中孔區(qū)的吸附過程均符合Intra-particle-diffusion模型,預(yù)示著吸附過程由粒內(nèi)擴(kuò)散過程控制,微孔區(qū)中的吸附過程由Homogeneous particle-diffusion模型控制;Freundlich模型更適合描述LZ-72對黃酮類化合物的熱力學(xué)吸附行為,說明LZ-72對虎杖中黃酮類化合物的吸附行為是多分子層物理吸附。
[Abstract]:Polygonum cuspidatum Sieb.et Zucc. (Polygonum cuspidatum) is one of the traditional Chinese medicinal materials in China. The 2000 edition of China pharmacopoeia is included in the first part. The bioactive components in the Polygonum cuspidatum are composed of Astragalus compounds and flavonoids. Modern pharmacological studies have shown that the physiological activity of resveratrol (Resveratrol) in the Astragalus is mainly manifested in the physiological activities of the Radix Polygoni cuspidatum. Antioxidative, anti free radicals, anti aging, anticancer, anti cardiovascular diseases, flavonoids (Flavonoids) have many medical effects, such as analgesic, antiviral, antilipidemic, hypoglycemic, antioxidation and other medical effects. At present, the research on the separation and purification of effective components from Polygonum cuspidatum has attracted more and more attention. This paper studies resveratrol from the Polygonum cuspidatum The main results are as follows: 1. The optimum extraction conditions of resveratrol from Polygonum cuspidatum were obtained by enzymatic hydrolysis. The optimum extraction conditions of resveratrol were optimized by single factor and orthogonal design. The extraction temperature was 45, the extraction time was 5 h, the amount of enzyme was 0.50% (m/m), the extract P H was 6.5, resveratrol. The extraction rate was 75.66%, the content was 3.80%, which was about 10 times higher than the content of the raw material (0.4%). Two, the purification effect of resveratrol from the crude extracts of the Polygonum cuspidatum was investigated by 32 kinds of MAR and its mixed bed (MARs). The results showed that the best purification effect of MARs LZ-94+LZ-78+LZ-76 to resveratrol was best, and the optimum adsorption condition was 0.. The concentration of adsorption solution was 0.. 25mg/m L, adsorption temperature 45 C, adsorption liquid volume 60.0m L, adsorption liquid p H=3.0, adsorption time 9 h; the optimum desorption conditions are: desorption temperature 55, ethanol volume fraction 70%, desorption time 2.5h, desorption solution P H=4.0. obtained MARs to resveratrol adsorption rate 97.96%, the desorption rate of 80.55%, the final recovery of resveratrol reached 78.60%, the purity of up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to 78.60%, purity up to the purity of up to the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity of up to 78.60%, the purity of the purity reached 78.60%. By 19.32%, the study on the adsorption kinetics of Resveratrol by 5.08 times.MARs to resveratrol shows that the adsorption of resveratrol conforms to the quasi two kinetic model. The adsorption process of the macroporous and mesoporous regions conforms to the Intra-particle-diffusion model, indicating that the adsorption process is controlled by the intragranular diffusion process and in the microporous region. The adsorption process is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of MARs on resveratrol. It shows that the adsorption behavior of resveratrol in the Polygonum cuspidatum by MARs is a multi molecular physical adsorption. Three, the purity is low after the separation and purification of resveratrol in the former part of the Polygonum cuspidatum. The extraction of Resveratrol by MARs was used in this part. First, the extraction method and process were studied. By comparing the extraction effects of different organic solvents and mixed solvents on resveratrol, it was found that the extraction effect of resveratrol was better when the organic solvent E/D=3/1 was used as the extractant, and the extraction of water phase and organic matter was obtained. The optimum volume ratio is Vw/Vo=2/3, the interval of extraction time is 60min, and the extraction 5 times is the most suitable. Under this condition, the extraction rate of resveratrol is 88.81%. The purity of the extraction method is 66.50%., which can guarantee the minimum decomposition loss of resveratrol under the premise that the resveratrol is fully hydrolyzed into resveratrol, and the E and D are both internationalized. The green solvents recognized by the academic community were environmentally friendly. After that, the samples were separated and purified by silica gel column chromatography and MARs technology, and two methods were compared. The recovery rate of resveratrol was 61.60% after the silica gel column chromatography was purified and the purity was 73.70%. After the purification of Resveratrol by MARs technology, the purified resveratrol was purified by silica gel column chromatography. The recovery rate of resveratrol is 81.10%, the purity is 87.12%., so the purification technology of MARs is better than silica gel column chromatography on the premise of purifying the same quality resveratrol. Four, the flavonoids in Polygonum cuspidatum are extracted by ultrasonic assisted method, and the extraction of flavonoids is optimized by single factor experiment and central combination design and response surface method. The optimum extraction conditions are as follows: the ethanol volume fraction of the extract is 60%, the extraction temperature is 45, the ultrasonic extraction time is 34 min, the ultrasonic power is 80W, the solid-liquid ratio 1/20 (g/m L), the optimum extraction times is 5 times. The extraction rate of flavonoids in the Polygonum cuspidatum is 94.50%, the purity of the initial extract is 31.00%. five, and 32 kinds of MAR and MA are investigated. Rs for the separation and purification of Flavonoids from the Polygonum cuspidatum, the result is that single MAR LZ-72 is the best for the separation and purification of flavonoids in the Polygonum cuspidatum. The best adsorption conditions are: the concentration of the original solution is 2.50mg/m L, the adsorption temperature is 30, and the adsorption liquid is 10% of the ethanol solution, the volume of the adsorbent is 60 m L, the adsorption time is 20 h, and the best desorption strip is the best. The results are as follows: the desorption temperature is 55 C, the solution volume is 110 m L, the volume fraction of ethanol is 90%, the desorption time is 20 h, the purity of the final product reaches 82.50%, the recovery rate reaches 89.70%.LZ-72 to the adsorption kinetics of flavonoids, which shows that the adsorption process is in accordance with the quasi two kinetics model, the adsorption process of the large pore and the mesoporous areas. According to the Intra-particle-diffusion model, the adsorption process is controlled by the intragranular diffusion process. The adsorption process in the microporous area is controlled by the Homogeneous particle-diffusion model, and the Freundlich model is more suitable for describing the thermodynamic adsorption behavior of LZ-72 to flavonoids, and the adsorption of flavonoids in the Polygonum cuspidatum by LZ-72 It is a physical adsorption of a multi molecular layer.
【學(xué)位授予單位】：蘭州理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O647.3;R284.2

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