本文選題：新疆紫草 + 大鼠��； 參考：《新疆醫(yī)科大學(xué)》2016年碩士論文

【摘要】：目的:建立檢測(cè)新疆紫草脂溶性成分的HPLC法,并應(yīng)用于體內(nèi)外藥物濃度的測(cè)定,考察新疆紫草脂溶性成分在體內(nèi)的處置情況。方法:(1)通過(guò)比較不同流動(dòng)相的分離效果及影響因素,建立了用于測(cè)定新疆紫草脂溶性成分體內(nèi)外濃度的HPLC法。(2)考察了方法學(xué)指標(biāo);(3)比較分析了不同有機(jī)溶劑萃取、酸化和堿化處理血液等方法對(duì)血中藥物萃取效果的影響。(4)考察了血漿與血清樣品的適用性和影響。(5)考察新疆紫草脂溶性成分在不同酸堿環(huán)境及模擬胃液、模擬腸液中的穩(wěn)定性試驗(yàn)。(6)給于大鼠灌胃給藥,分別于給藥前及給藥后不同時(shí)間采血,分離血清和血漿,收集給藥前及給藥后的48小時(shí)內(nèi)的尿液和糞便,進(jìn)行HPLC分析,測(cè)定這些樣品中左旋紫草素、乙酰紫草素、去氧紫草素、β-β二甲基丙烯酰紫草素等單一成分的濃度。結(jié)果:(1)色譜條件為:色譜柱COSMOIL C18-AR-Ⅱ譜柱(250mm×4.6mm,5um);流速:1.0m L/min;柱溫:(25)℃;檢測(cè)波長(zhǎng):275nm,流動(dòng)相A液-乙腈,B液-甲酸:水(0.5:300)(v/v)溶液,進(jìn)樣量20μl,梯度洗脫,0~10min(60-40)%;15~30min(70-30)%;30~35min(60-40)%;檢測(cè)時(shí)間35min。在該色譜條件下,新疆紫草脂溶性提取物中左旋紫草素、乙酰紫草素、β-β二甲基丙烯酰紫草素得到較好的分離。(2)方法學(xué)指標(biāo)符合規(guī)定:日間精密度試驗(yàn)中各目標(biāo)峰保留時(shí)間RSD1%,樣品12h內(nèi)穩(wěn)定相對(duì)峰面積RSD3%,重復(fù)性試驗(yàn)相對(duì)峰面積RSD3%;(3)血漿和血清樣品中藥物提取分離條件為,樣品先用等體積的稀鹽酸酸化,用乙酸乙酯萃取3次,乙腈沉淀蛋白。血漿樣品在5μg/ml~500μg/m濃度范圍內(nèi)與峰面積呈良好線性關(guān)系,回歸方程為左旋紫草素:y=20.613x+979.88;乙酰紫草素:y=24.254x-57.209;β-β二甲基丙烯酰紫草素:y=48.406x+3180.5;相關(guān)系數(shù)分別為0.997、0.9957、0.9935。(4)樣品的適用性方面,血漿樣品較適合于疆紫草脂溶性提取物成分的測(cè)定,在血清樣品中添加適量的肝素后,峰形和峰面積得到了改善。(5)新疆紫草脂溶性成分(左旋紫草素、乙酰紫草素、β-β二甲基丙烯酰紫草素)在pH2.5磷酸鹽緩沖液及人工胃液中相對(duì)穩(wěn)定,但隨著時(shí)間的延長(zhǎng),含量呈緩慢下降趨勢(shì);在pH5.8磷酸鹽緩沖液呈明顯的下降趨勢(shì);隨著pH值的上升,含量下降迅速。(6)經(jīng)大鼠灌胃給藥后0~48小時(shí)內(nèi)血樣、尿液中均未檢測(cè)到新疆紫草脂溶性提取物所含有原型物質(zhì)特有峰。而在糞便中發(fā)現(xiàn)有左旋紫草素、乙酰紫草素、β-β二甲基丙烯酰紫草素等成分的峰。結(jié)論:大鼠灌胃后的血液樣品及尿液中未檢測(cè)到新疆紫草脂溶性提取物,而在糞便發(fā)現(xiàn)有該提取物的相關(guān)成分,說(shuō)明新疆紫草脂溶性提取物在胃腸道幾乎不吸收或吸收很少,其吸收、分布、代謝、排泄過(guò)程有待于進(jìn)一步深入研究。
[Abstract]:Objective: to establish a HPLC method for the determination of liposoluble components in Xinjiang, and to determine the treatment of liposoluble components in Xinjiang in vivo. Methods: (1) a HPLC method was established to determine the concentration of liposoluble constituents in Xinjiang by comparing the separation effect and influencing factors of different mobile phases. (2) the methodological indexes were investigated, and (3) the effects of different organic solvent extraction, acidification and alkaline treatment of blood on the effect of drug extraction in blood were compared and analyzed. (4) the applicability and influence of plasma and serum samples were investigated. (5) the stability of lipid soluble fractions in different acid base environment and simulated gastric juice in Xinjiang and the stability of simulated intestinal fluid were investigated. (6) (6) the rats were given the medicine by gavage, the blood was collected before and after the administration, the serum and plasma were separated, the urine and feces were collected for 48 hours before and after the administration, and the HPLC analysis was carried out to determine the single formation of levoshikonin, acetoxin, deoxyshioxin, beta - beta two methacyleylephrin, etc. Results: (1) the chromatographic conditions are: chromatography column COSMOIL C18-AR- II column (250mm x 4.6mm, 5um); flow rate: 1.0m L/min; column temperature: (25) C; detection wavelength: 275nm, liquid phase A liquid acetonitrile, B liquid formic acid: water (0.5:300) (v/v) solution, the sample volume is 20 mu, gradient elution, 60-40%; 70-30%; 60-40%; detection time Under this chromatographic condition, a better separation was obtained from levopurple, acetyl, beta two methacrolein in the liposoluble extracts of Xinjiang purple grass. (2) the methodological indexes were in accordance with the regulations: the retention time of each target peak in the day precision test was RSD1%, the stable relative peak area within the sample 12h was RSD3%, and the relative peak area of the repeatability test was RSD3%; 3) the extraction and separation condition of plasma and serum samples is that the sample is acidified by dilute hydrochloric acid with equal volume first, extract 3 times with ethyl acetate and precipitate acetonitrile protein. The plasma sample has a good linear relationship with the peak area in the range of 5 mu g/ml~500 g/m, and the regression equation is levoviolet herb: y=20.613x+979.88; acetyl purplicin: y=24.254x-57.209 The beta beta two methacryl purple herb: y=48.406x+3180.5; the correlation coefficient is 0.997,0.9957,0.9935. (4), respectively. The plasma sample is suitable for the determination of the composition of the liposoluble extract of the purple herb. After adding a proper amount of heparin in the serum samples, the peak shape and peak area have been improved. (5) the liposoluble composition of Xinjiang purple grass (left) It was relatively stable in pH2.5 phosphate buffer solution and artificial gastric juice, but the content decreased slowly in pH2.5 phosphate buffer solution and artificial gastric juice, while pH5.8 phosphate buffer solution decreased obviously with the rise of pH value. (6) 0~48 was small after gavage of rats. In the urine and urine, there were no specific peaks in the lipid soluble extracts of Xinjiang herb, and the peaks were found in the feces, such as levopurple, acetylenetin, beta - beta two methacryrodacysin. Conclusion: the blood samples and urine of rats after the gavage of the stomach were not detected in the liposoluble extract of Xinjiang. The related components of the extract were found in the feces, indicating that the liposoluble extract of Xinjiang herb was hardly absorbed or absorbed in the gastrointestinal tract, and its absorption, distribution, metabolism, and excretion process need to be further studied.

【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R29;O657.72

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