本文選題：牛初乳 + 功能乳寡糖�。� 參考：《華東理工大學(xué)》2017年碩士論文

【摘要】：牛初乳中含有種類豐富且具有生理活性的功能寡糖,在食品和保健品領(lǐng)域有潛在的應(yīng)用前景。目前對(duì)這些寡糖的研究尚不充分,對(duì)于結(jié)構(gòu)復(fù)雜、種類較多的混合寡糖缺乏有效的分離方法,極大地限制了牛初乳中功能寡糖的開發(fā)和應(yīng)用。本論文旨在利用膜分離耦合色譜分離技術(shù)對(duì)牛初乳中的功能寡糖特別是唾液酸化的混合寡糖進(jìn)行分離純化,優(yōu)化并得到最佳的分離工藝參數(shù),獲得高純度的唾液酸乳糖,對(duì)牛初乳的高值化利用提供依據(jù)。首先,本研究建立了一種快速、準(zhǔn)確的分析測定牛初乳中糖類物質(zhì)的方法;并通過乳糖、半乳糖、3'-唾液酸乳糖、6'-唾液酸乳糖四種牛乳中含量較多的糖類標(biāo)準(zhǔn)品對(duì)該方法進(jìn)行驗(yàn)證。結(jié)果表明:乳糖、半乳糖、3'-唾液酸乳糖、6'-唾液酸乳糖,分別在0.03-5.00 mg/mL、0.02-5.00 mg/mL、0.05-2.00 mg/mL、0.05-2.00 mg/mL 濃度范圍內(nèi),該方法具有良好的線性關(guān)系(R20.99)和重復(fù)性(峰面積的RSD4%,保留時(shí)間RSD2%);四種標(biāo)準(zhǔn)品的回收率均在100%-115%之間,LOD、LOQ分別為9 μg/mL、30 μg/mL,7 μg/mL、20μg/mL,125μg/mL、200μg/mL,350μg/mL、450μg/mL�；诮⒌臋z測方法對(duì)牛初乳中主要寡糖組分進(jìn)行了分析測定,發(fā)現(xiàn)牛初乳中大約有27種不同的功能寡糖,其含量大約為1.93 g/L。其次,根據(jù)牛初乳組成成分的物理化學(xué)特性,選擇了截留分子量為10 kDa的超濾膜和截留分子量為280 Da的納濾膜對(duì)牛初乳中的寡糖進(jìn)行初步分離純化。結(jié)果表明:超濾過程中壓力和溫度對(duì)膜通量具有明顯的影響,但對(duì)蛋白質(zhì)截留率和總糖的回收率無明顯作用。在最適的操作壓力和操作溫度(5 bar,40℃)條件下蛋白質(zhì)的截留率為96.0%,總糖的回收率為98.3%。納濾分離過程中膜通量隨著壓力和溫度的增大明顯的變大,單糖和乳糖的截留率隨著壓力的增大先升高后降低,隨著溫度的升高不斷的降低;而壓力和溫度的變化對(duì)寡糖的截留率無明顯的影響。在最適的操作壓力和操作溫度(8 bar,45℃)條件下,單糖和乳糖的截留率為41.9%,寡糖的截留率為97.0%。最后,對(duì)膜分離得到的寡糖粗提液的成分進(jìn)行測定,發(fā)現(xiàn)乳糖和唾液酸乳糖含量最高。根據(jù)乳糖和唾液酸乳糖的性質(zhì)差異及色譜分離技術(shù)的原理,選擇D315、QY-P001、QY-Z001、PCR642Ca、CGC100X8五種樹脂作為研究對(duì)象,綜合乳糖和唾液酸乳糖的分離度、回收率和保留時(shí)間等因素,篩選合適的樹脂。結(jié)果表明:QY-Z001型陽離子交換樹脂是最適合分離唾液酸乳糖和乳糖的樹脂,且在該樹脂上乳糖為強(qiáng)保留組分,唾液酸乳糖為弱保留組分。以QY-Z001樹脂作為固定相,去離子水作為流動(dòng)相,研究了徑高比、柱溫、洗脫速度、進(jìn)樣濃度等因素對(duì)乳糖和唾液酸乳糖分離效果的影響。結(jié)果表明:色譜柱徑高比為1:35、溫度為55℃、流速1.0 mL/min、寡糖混合物濃度為100 mg/mL的條件下,寡糖混合物中唾液酸乳糖和乳糖的分離效果最好,分離度可以達(dá)到0.96;色譜分離得到的樣品中唾液酸乳糖和乳糖的純度分別為84.6%、99.3%,回收率分別為76.9%、100.0%。樹脂QY-Z001適合牛初乳寡糖混合物中唾液酸乳糖和乳糖的分離,且具有良好的分離效果。研究結(jié)果為牛初乳中功能寡糖的分離純化提供了理論依據(jù)。
[Abstract]:Bovine colostrum contains a variety of functional oligosaccharides with physiological activity, have potential applications in food and health care products. The study of these oligosaccharides is not sufficient, for the complex structure, many types of mixed oligosaccharides lack effective separation methods, which greatly limits the development and application of functional oligosaccharides in bovine colostrum this paper aims to use membrane separation. The coupling function of chromatographic separation of bovine colostrums oligosaccharides in particular hybrid oligosaccharide sialyl were purified and separated, optimize the best technology parameter and obtain high purity sialyllactose, bovine colostrum high value utilization is provided. Firstly, this study established a rapid the analysis method, accurate determination of carbohydrates in bovine colostrum; and by lactose, galactose, 3'- sialyllactose, 6'- sialyllactose four milk content of more sugar The standard to verify the method. The results showed that: lactose, galactose, 3'- sialyllactose, 6'- sialyllactose, respectively, in 0.03-5.00 mg/mL, 0.02-5.00 mg/mL, 0.05-2.00 mg/mL, 0.05-2.00 mg/mL concentration range, the method has a good linear relationship (R20.99) and repeatability (peak area of RSD4%, retention time RSD2%); four standard recovery rate was between 100%-115%, LOD and LOQ were 9 g/mL, 30 g/mL, 7 g/mL, 20 g/mL, 125 g/mL, 200 g/mL, 350 g/mL, 450 g/mL. based on the detection method of primary bovine milk oligosaccharides group by analysis, found that there are about 27 different species of functional oligosaccharides in bovine colostrum, the content of about 1.93 g/L. second, according to the physical and chemical properties of bovine colostrum composition, the molecular weight cut-off of 10 kDa membrane with a MWCO of 280 Da nanofiltration membrane in bovine colostrum Oligosaccharides were purified. The results showed that the ultrafiltration pressure and temperature has obvious influence on the membrane flux, but the rate has no significant effect on the recovery of protein retention rate and total sugar. The operating pressure and the optimum temperature (5 bar, 40 C) under the condition of protein retention rate was 96%. The total sugar recovery rate of 98.3%. in the process of nanofiltration membrane flux with temperature and pressure increased significantly larger, monosaccharide and lactose retention rate with the increase of pressure increased first and then decreased, with the increase of temperature decreasing; and the pressure and temperature of the oligosaccharides retention rate had no significant effect. In the operating pressure and temperature optimum (8 bar, 45 DEG C) condition, monosaccharide and lactose retention rate of 41.9%, oligosaccharides retention rate is 97.0%. finally, the membrane separation obtained oligosaccharide crude extract components were measured and found that lactose and sialic acid The highest content of lactose and lactose. According to the differences between the sialyllactose and the chromatographic separation technology principle, select D315, QY-P001, QY-Z001, PCR642Ca, CGC100X8 five resin as the research object, synthesis of lactose and sialyllactose separation factors, the recovery rate and retention time, selecting the appropriate resin. The results showed that: QY-Z001 cation exchange resin is the most suitable resin for separation of sialyllactose and lactose, and the resin for lactose retention components, sialyllactose for weak retention components. Using QY-Z001 resin as stationary phase, deionized water as mobile phase, the ratio of diameter to height, column temperature, elution speed, influence the separation effect the sample concentration of lactose and sialyllactose. The results showed that the column ratio is 1:35, the temperature is 55 DEG C, the flow rate of 1 mL/min, the concentration of a mixture of 100 mg/mL, oligosaccharide mixture in saliva The best separation effect of acid lactose and lactose, the separation degree can reach 0.96; the purity of chromatographic separation of sialyllactose and lactose samples obtained were 84.6%, 99.3%, the recovery rate was 76.9%, 100.0%. separation resin QY-Z001 for sialyllactose and lactose in bovine colostrum oligosaccharide mixture, and has a good separation effect. The research results provide a theoretical basis for the separation function of bovine colostrum in the purification of oligosaccharides.

【學(xué)位授予單位】：華東理工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TS252.1

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