本文選題：葛根 + 葛花��； 參考：《長(zhǎng)春師范大學(xué)》2017年碩士論文

【摘要】：本文以葛根、葛花和大豆為研究對(duì)象,利用高速逆流色譜(High-speed Counter-current Chromatography,HSCCC)和半制備高效液相色譜(Semi-preparative High-performance Liquid Chromatography,SPHPLC)對(duì)中藥材中的異黃酮類(lèi)成分進(jìn)行提取分離,采用超濾及PC12細(xì)胞技術(shù)對(duì)體外活性進(jìn)行研究。葛根為藥食兩用植物,是野葛的干燥根。本文應(yīng)用加熱回流技術(shù)對(duì)葛根提取工藝進(jìn)行研究,采用單因素實(shí)驗(yàn)結(jié)合正交實(shí)驗(yàn)優(yōu)化提取工藝。確定最佳提取工藝為:提取溶液為80%乙醇溶液、提取次數(shù)2次、料液比為l:20,提取時(shí)間120 min,獲得了葛根異黃酮總提取物。以大豆苷元的提取量為目標(biāo),提取率達(dá)到1.263 mg/g。以乳酸脫氫酶作為生物靶分子,以超濾質(zhì)譜技術(shù)作為篩選方法篩選酶抑制劑,確定化合物大豆苷元具有較強(qiáng)的酶抑制活性。以高速逆流色譜技術(shù)作為分離方法,以石油醚-乙酸乙酯-甲醇-水(5:5:4:5,v:v:v:v)作為溶劑系統(tǒng),分離得到的大豆苷元純度為99.2%。葛花為豆科植物野葛的花。本文以加熱回流技術(shù)對(duì)葛花提取工藝進(jìn)行研究,采用單因素實(shí)驗(yàn)結(jié)合正交實(shí)驗(yàn)優(yōu)化提取工藝。確定最佳提取工藝為:提取溶液為70%乙醇溶液、提取次數(shù)3次、料液比l:15,提取時(shí)間120 min,獲得了葛花異黃酮總提取物。以鳶尾苷的提取量為目標(biāo),提取率達(dá)到5.601 mg/g。以乳酸脫氫酶作為生物靶分子,以超濾質(zhì)譜技術(shù)作為篩選方法篩選酶抑制劑,并通過(guò)MTT比色法對(duì)分離得到的成分進(jìn)行體外抗腦卒中活性測(cè)定,確定化合物鳶尾黃素-7-O-木糖葡萄糖苷(tectorigenin-7-O-xylosylglucoside)、鳶尾苷(tectoridin)、鳶尾苷元(tectorigenin)具有較強(qiáng)的酶抑制活性。利用半制備高效液相色譜對(duì)上述三種乳酸脫氫酶抑制劑進(jìn)行分離,共分離得到3個(gè)化合物,純度分別為鳶尾黃素-7-O-木糖葡萄糖苷90.1%,鳶尾苷93.4%,鳶尾苷元95.5%。通過(guò)MTT實(shí)驗(yàn)進(jìn)一步驗(yàn)證了葛花總提取物可以保護(hù)MPP+損傷的PC 12細(xì)胞。大豆為豆科大豆屬草本植物。本文利用加熱回流技術(shù)對(duì)大豆中的異黃酮類(lèi)成分進(jìn)行提取,以乳酸脫氫酶作為生物靶分子,以超濾質(zhì)譜技術(shù)作為篩選方法篩選酶抑制劑,對(duì)其活性進(jìn)行評(píng)價(jià)。確定大豆中有六種化合物為乳酸脫氫酶抑制劑,其抑制率分別為:乙酰大豆黃素糖苷(48.36%),丙二酰染料木素糖苷(47.14%),大豆苷(41.92%),黃豆黃苷(39.13%),染料木苷(28.41%),乙酰黃豆黃素糖苷(24.21%)。通過(guò)MTT實(shí)驗(yàn)進(jìn)一步驗(yàn)證了大豆總提取物對(duì)缺血性腦損傷有保護(hù)作用。
[Abstract]:In this paper, the isoflavones from Radix Puerariae, Radix Puerariae and Soybean were extracted and separated by High-speed Counter-current Chromatography-HSCCC and semi-preparative High-performance Liquid Chromatography-SPHPLCc. In vitro activity was studied by ultrafiltration and PC12 cell technology. Pueraria lobata is a medicinal and food plant, is the dry root of Pueraria lobata. In this paper, the extraction technology of Pueraria lobata was studied by heating reflux technique, and the extraction process was optimized by single factor experiment and orthogonal experiment. The optimum extraction technology was determined as follows: the extraction solution was 80% ethanol solution, the extraction times were 2 times, the ratio of material to liquid was 1: 20, the extraction time was 120 min, and the total extract of isoflavone from Pueraria lobata was obtained. The extraction rate of daidzein was 1.263 mg / g. Lactate dehydrogenase (LDH) was used as biological target molecule and ultrafiltration mass spectrometry (UF / MS) was used to screen enzyme inhibitors. It was confirmed that daidzein had strong enzyme inhibitory activity. The purity of daidzein was 99.2 using high speed countercurrent chromatography and petroleum ether-ethyl acetate methanol-water 5: 5: 5: 5: 5: 5: v: v: v) as solvent system. Pueraria lobata is the flower of legume. In this paper, the extraction technology of Pueraria lobata was studied by heating reflux technique, and the single factor experiment and orthogonal experiment were used to optimize the extraction process. The optimum extraction technology was determined as follows: the extraction solution was 70% ethanol solution, the extraction times were 3 times, the ratio of material to liquid was 1: 15, the extraction time was 120 min, and the total extract of isoflavone from Pueraria lobata was obtained. The extraction rate of Iris was 5.601 mg / g. Lactate dehydrogenase (LDH) was used as biological target molecule, ultrafiltration mass spectrometry (UF MS) was used as screening method to screen enzyme inhibitors, and the antistroke activity of the isolated components was determined by MTT colorimetry in vitro. It was confirmed that the compounds of Iridin-7-O- xylosylglucoside, tectorigenin-7-O-xylosylglucoside, Iris tectoridinanum and tectorigenin have strong enzyme inhibitory activity. Three kinds of lactate dehydrogenase inhibitors were separated by semi-preparative high performance liquid chromatography. The purity of the three compounds were Iris flavin-7-O-xylose glucoside 90.1, iris 93.4 and iris 95.5 respectively. It was further proved by MTT experiment that total extract of Pueraria lobata could protect PC12 cells damaged by MPP. Soybean is a kind of herbaceous plant of Leguminosae. In this paper, isoflavones from soybean were extracted by heating reflux technique. Lactate dehydrogenase (LDH) was used as biological target molecule and ultrafiltration mass spectrometry (UF MS) was used to screen enzyme inhibitors. Six compounds in soybean were identified as lactate dehydrogenase inhibitors, the inhibition rates were: acetyldaidzein 48.36, malonyl genistein 47.1414, daidzein 41.92, daidzein 39.13, genistein 28.41, acetyl daidzein 24.2121. The inhibitory rates were as follows: acetyldaidzein 48.36, malonyl genistein 47.1421, daidzein 41.92, daidzein 39.13, genistein 28.41, acetyldaidzein 24.2121. The protective effect of total soybean extract on ischemic brain injury was further verified by MTT test.
【學(xué)位授予單位】：長(zhǎng)春師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R284;R285;O652.63

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本文編號(hào)：1785902