【摘要】：[目的]優(yōu)化雨生紅球藻有機溶劑提取工藝,尋找最佳的萃取條件,精確檢測雨生紅球藻萃取物中的蝦青素酯含量,探索適用于大規(guī)模工業(yè)生的高速、高效、低成本提取方法;建立小鼠糖尿病模型,并給予不同濃度的雨生紅球藻源蝦青素酯,研究其對糖尿病小鼠糖耐量的影響、抗氧化指標的影響、血脂的調(diào)節(jié)情況及胰腺組織的損傷修復情況。探討蝦青素酯改善糖尿病及其并發(fā)癥的作用機制,并驗證高劑量蝦青素酯對正常小鼠的影響;結合市場需求及原料特性,制備以雨生紅球藻萃取物為原料的功能食品。[方法]1.從原料的預處理入手,采用物理、化學處理相結合的手段,首先將雨生紅球藻微藻進行破壁粉碎,以食品級有機溶劑乙酸乙酯作為萃取劑,應用SPSS 22.0統(tǒng)計學軟件,對萃取溫度(40℃、50℃、60℃)、原料顆粒度(40目、80 目、160 目)、料液比(1:4、1:8、1:16)、萃取時間(30min、60min、90min)設計正交實驗,并對單因素水平進行驗證,通過與仲裁法丙酮萃取得率作對比,結合實際操作,確定雨生紅球藻最佳萃取因素水平。2.運用高效液相色譜法(High Performance Liquid Chromatography HPLC),精確檢測雨生紅球藻萃取物中蝦青素酯的含量。3.將喂食30 d高劑量蝦青素酯正常小鼠,與喂食30 d溶劑的正常小鼠血糖值做對比,用SPSS 22.0統(tǒng)計學軟件進行分析,比較兩組小鼠的血糖值,分析蝦青素酯對正常小鼠血糖值的影響。4.采取四氧嘧啶腹腔注射的方式建立糖尿病小鼠模型,設立高中低三個劑量蝦青素酯組和二甲雙胍、維生素C兩個陽性藥對照組,給藥期周期為30 d,每7 d記錄1次小鼠體重;給藥期末進行耐糖量實驗,利用血糖儀測定血糖值,運用全自動生化分析儀和相關試劑盒檢測丙二醛(MDA)、谷胱甘肽(GSH-Px)、超氧化物歧化酶(T-SOD)等抗氧化指標,總膽固醇(CHO)、甘油三酯(TG)等血脂指標,并制作胰腺病理切片,觀察胰島組織形態(tài)變化。5.根據(jù)原料理化性質(zhì),選用軟膠囊作為蝦青素酯功能食品最終產(chǎn)品劑型,并進行小試試驗研究生產(chǎn)。[結果]1.萃取溫度40℃,原料顆粒度160目,料液比1:16,萃取時間90min,此條件下雨生紅球藻萃取效果最好,萃取率高達90.8%(W/W)。2.通過HPLC分析,確定雨生紅球藻萃取物中蝦青素酯含量為5.46%。3.高劑量蝦青素酯喂養(yǎng)30 d,對正常小鼠的血糖無影響。4.四氧嘧啶造模后小鼠毛色雜亂暗淡,出現(xiàn)明顯多飲、多食、多尿的"三多"癥狀,與正常組小鼠相比,體重明顯減輕(P0.05)。給予不同劑量蝦青素酯喂養(yǎng)30 d后,各給藥組體重平均值與模型對照組相比有所上升,低劑量蝦青素酯組小鼠體重升高明顯,存在統(tǒng)計學差異(P0.05)。高、中、低劑量蝦青素酯組的空腹血糖值有不同程度下降,與模型對照組比較有統(tǒng)計學差異(P0.05,P0.05,P0.05),且有一定的量效關系,但降血糖效果不如陽性對照藥二甲雙胍。在口服葡萄糖耐量實驗中,各組小鼠的糖負荷均在30 min時達到峰值,高劑量蝦青素酯組和中劑量蝦青素酯組血糖值在120 min時明顯低于對照組(P0.05,p0.05),高、中劑量蝦青素酯可以提高糖尿病小鼠對外源性葡萄糖的耐受力,與對照組相比有統(tǒng)計學差異。計算血糖線下面積,高、中、低劑量蝦青素酯組均比模型對照組低,且有統(tǒng)計學意義(P0.05,P0.05,P0.05),表明蝦青素酯可降低各時間點血糖,但效果不如陽性對照藥二甲雙胍�？寡趸笜�,與模型對照組相比,高、中、低劑量蝦青素酯組血清MDA含量顯著下降(P0.01,P0.01,P0.05),血清GSH-Px活性單位含量、血清T-SOD活性單位含量顯著上升(P0.01,P0.01,P0.01),并存在著一定的量效關系,抗氧化指標結果表明:蝦青素酯具有顯著的抗氧化作用,且明顯高于陽性對照藥維生素C。蝦青素各劑量組血清CHO、TG含量明顯低于對照組,差異具有統(tǒng)計學意義(P0.01,P0.01,P0.01),提示蝦青素酯具有調(diào)節(jié)血脂的作用。5.以雨生紅球藻萃取物為主要原料,葵花籽油為輔料,制成內(nèi)容物0.5 g的蝦青素軟膠囊。
[Abstract]:[Objective] to optimize the extraction process of the organic solvent of rhodotrazina, find the best extraction conditions, accurately detect the content of astaxanthin ester in the extract of rhodotrae alga, explore the high speed, high efficiency and low cost extraction method suitable for large-scale industrial students, establish a diabetic model of mice, and give astaxanthin esters of different concentrations of the raining red ball algae. The influence of the glucose tolerance, the influence of antioxidant index, the regulation of blood lipid and the injury and repair of pancreatic tissue were studied. The mechanism of astaxanthin ester to improve diabetes and its complications was investigated, and the effect of high dose astaxanthin ester on normal mice was tested and the effect of astaxanthin ester on normal mice was verified. [method]1., starting with the pretreatment of the raw material, starting with the pretreatment of raw materials, using the means of physical and chemical treatment, the rain raw red alga microalgae were broken, and the organic solvent ethyl acetate was used as the extractant and the SPSS 22 system software was applied to the extraction temperature (40, 50, 60), and the raw materials were used. Granularity (40 orders, 80 orders, 160 orders), material liquid ratio (1:4,1:8,1:16), extraction time (30min, 60min, 90min) design orthogonal experiment, and verify the single factor level, by comparing with the arbitration method of acetone extraction, combining actual operation, to determine the optimum extraction factor level of raococcus red pellet using high performance liquid chromatography (High Performance L) method (High Performance L). Iquid Chromatography HPLC), the content of astaxanthin ester in the extract of rhodotrae alga was accurately measured by.3., 30 d high dose astaxanthin ester was fed to normal mice, and the normal mice fed with 30 d solvent were compared. The blood glucose of two groups of mice was compared with the SPSS 22 statistical software, and the blood of astaxanthin was compared to the normal mice blood. The effect of sugar value on.4. was established by intraperitoneal injection of four pyrimidine to establish a diabetic mouse model. Three doses of astaxanthin ester group and metformin and two vitamin C positive drug control groups were set up in high school. The period of the administration period was 30 d, and 1 mice were recorded every 7 d; the glucose tolerance test was carried out at the end of the period and the blood glucose was measured by blood glucose meter, The antioxidant indexes such as malondialdehyde (MDA), glutathione (GSH-Px), superoxide dismutase (T-SOD), total cholesterol (CHO), triglyceride (TG) and other lipid indexes were detected by the automatic biochemical analyzer and related kit. The pathological sections of pancreas were made and the morphological changes of pancreatic islets were observed and.5. was used as the shrimp green according to the properties of raw material. The final product form of vegetarian ester function food was produced in a small test. [results]1. extraction temperature 40 C, raw material granularity 160 mesh, material liquid ratio 1:16, extraction time 90min, the extraction efficiency is best, the extraction rate is up to 90.8% (W/W).2. through HPLC analysis, determine the astaxanthin ester content in the raining red pellet extract is 5 .46%.3. high dosage of astaxanthin ester was fed to 30 d, and the blood sugar of normal mice had no effect on the hair color and confusion of mice after the model of.4. four oxacil. The "more than 3" symptoms of polyuria, polyuria and polyuria were obviously reduced (P0.05). The average weight and model of body weight of each group were given after 30 d were fed with different doses of astaxanthin ester. Compared with the control group, the body weight of the low dose astaxanthin ester group was significantly higher than that of the low dose astaxanthin ester group (P0.05). High, middle and low dosage of astaxanthin ester group decreased the fasting blood glucose in different degrees, compared with the model control group, there was a statistical difference (P0.05, P0.05, P0.05), and there was a certain dose effect relationship, but the effect of hypoglycemic effect was not as good as that of Yang. In the oral glucose tolerance test, in the oral glucose tolerance test, the glucose load reached the peak at 30 min. The high dose of astaxanthin ester group and the middle dose astaxanthin ester group were significantly lower than the control group (P0.05, P0.05) at 120 min. High dosage of astaxanthin ester could improve the tolerance to exogenous glucose in diabetic mice. The area under the blood glucose line, high, middle and low dosage of astaxanthin ester group were lower than the model control group, and there were statistical significance (P0.05, P0.05, P0.05), indicating that astaxanthin ester could reduce the blood glucose at all time points, but the effect was not as effective as the positive metformin. The antioxidant index was compared with the model control group. The content of serum MDA in high, middle and low dose astaxanthin ester group decreased significantly (P0.01, P0.01, P0.05), the content of GSH-Px active unit in serum, the content of serum T-SOD active unit increased significantly (P0.01, P0.01, P0.01), and there was a certain quantitative effect relationship. The antioxidant index showed that astaxanthin ester had significant antioxidant effect and was obviously higher than positive. The content of serum CHO and TG in each dose group of vitamin C. and astaxanthin was significantly lower than that of the control group. The difference was statistically significant (P0.01, P0.01, P0.01), suggesting that astaxanthin ester had the effect of regulating blood lipids,.5. was the main raw material, and sunflower seed oil was used as the auxiliary material, and the astaxanthin soft capsule of 0.5 g was made.
【學位授予單位】：中央民族大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：TS201.4

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