本文關(guān)鍵詞： 牦牛軟骨 提取 純化 響應(yīng)面分析 生物活性　出處：《西北民族大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目前國內(nèi)外主要從牛、豬及鯊魚等動物軟骨組織中提取硫酸軟骨素,且提取工藝粗糙,產(chǎn)品純度較低,質(zhì)量較差。本文以天祝牦牛軟骨為原料,優(yōu)化軟骨中硫酸軟骨素的提取、純化等工藝流程,并研究硫酸軟骨素的免疫增強(qiáng)效果、降血脂功能和抗凝血作用,為提高牦牛副產(chǎn)品的深加工和利用價(jià)值進(jìn)行了有益的探索。主要試驗(yàn)及結(jié)論如下:1、牦牛硫酸軟骨素的提取方法優(yōu)化。以硫酸軟骨素純度為考察指標(biāo),對稀堿-酶解法、堿鹽-酶解法和超聲波輔助堿鹽-酶解法等三種提取方法進(jìn)行比較篩選,并通過單因素及響應(yīng)面分析確定超聲波輔助堿鹽提取硫酸軟骨素的最佳工藝條件為:料液比為1:6.5(g/mL)、堿液濃度為8%、提取時(shí)間為2.7h、Na+濃度為2.5mol/L、提取溫度為40℃,CS純度平均值為55.60%。對木瓜蛋白酶酶解工藝進(jìn)行單因素及響應(yīng)面分析優(yōu)化,確定木瓜蛋白酶酶解的最佳工藝為:酶解溫度為36.5℃,酶解pH為6.9,加酶量為1:829,酶解時(shí)間為1.5h,CS純度平均值為71.60%。2、牦牛硫酸軟骨素的陰離子交換樹脂法純化工藝優(yōu)化。最佳工藝條件為:樹脂與CS粗提物水溶液體積比為1:1,CS粗提物水溶液濃度為25mg/mL,CS粗提物水溶液流速為1.OmLmmin,吸附時(shí)間70min,吸附后將CS粗提物水溶液流出,保持樹脂與NaCl溶液體積比為1:5,NaCl溶液濃度為2.5mo]/L,NaCl溶液流速為1.0mL/min,解吸時(shí)間90min。純化物含量結(jié)果平均為82.77%。然后選擇葡聚糖凝膠G75進(jìn)行再次純化,得到的硫酸軟骨素的純度為99.30%。3、牦牛硫酸軟骨素生物活性作用研究。對牦牛硫酸軟骨素進(jìn)行免疫增強(qiáng)作用的研究,結(jié)果表明:高劑量硫酸軟骨素對小鼠血清溶血素有正影響,且低、中、高劑量的硫酸軟骨素對脾臟指數(shù)及胸腺指數(shù)都有所提高,對小鼠體液免疫功能有增強(qiáng)作用。對牦牛硫酸軟骨素進(jìn)行降血脂功能的研究,結(jié)果表明:對高血脂癥小鼠進(jìn)行灌胃硫酸軟骨素后,可降低小鼠血液中的總膽固醇、甘油三酯、低密度脂蛋白指標(biāo),同時(shí)升高高密度脂蛋白指標(biāo)。對牦牛軟骨中提取的CS進(jìn)行抗凝血作用的研究。結(jié)果顯示CS具有一定的抗凝血作用,但其作用遠(yuǎn)低于肝素。
[Abstract]:At present, chondroitin sulfate is extracted from chondroitin sulfate from cartilage tissue of cattle, pig and shark at home and abroad, and the extraction process is rough, the purity of the product is low and the quality is poor. This article takes the cartilage of Tianzhu yak as raw material. The extraction and purification of chondroitin sulfate from cartilage were optimized, and the immune enhancement, blood lipid lowering and anticoagulant effect of chondroitin sulfate were studied. In order to improve the value of further processing and utilization of yak by-products, the main experiments and conclusions are as follows: 1. The extraction method of chondroitin sulfate from yak was optimized. The purity of chondroitin sulfate was taken as the index. Three extraction methods, namely dilute alkali-enzymatic hydrolysis, alkaline salt-enzyme hydrolysis and ultrasonic assisted alkali-enzymatic hydrolysis, were compared and screened. The optimum conditions of ultrasonic assisted extraction of chondroitin sulfate by single factor and response surface analysis were as follows: the ratio of material to liquid was 1: 6.5g / mL, and the concentration of alkali was 8%. The extraction time was 2.7 h, the concentration of Na was 2.5 mol / L, and the extraction temperature was 40 鈩，

本文編號：1475317