本文關(guān)鍵詞：毛細管電泳—電化學檢測在生物胺及中草藥活性成分測定中的應(yīng)用研究，由筆耕文化傳播整理發(fā)布。

        第一章緒論毛細管電泳(Capillary Electrophoresis, CE),是一種發(fā)展迅速的新型分離分析技術(shù),它以毛細管為分離通道,以高壓直流電場為驅(qū)動力,根據(jù)被分析物特性的差異實現(xiàn)分離分析的技術(shù),具有分離效率高、速度快、樣品用樣量少、自動化程度高等優(yōu)點。CE的應(yīng)用得到了飛速發(fā)展,目前毛細管電泳已被廣泛用于蛋白質(zhì)、氨基酸、無機離子、有機化合物、藥物的分離分析,涉及食品化學、藥物化學、分析化學、臨床化學、環(huán)境化學等諸多學科領(lǐng)域。緒論簡要回顧了CE技術(shù)的發(fā)展歷程,介紹了其原理、分離模式、進樣方式和檢測技術(shù)特點,并著重介紹了CE與電化學檢測器聯(lián)用技術(shù),特別是安培檢測(AD)技術(shù)和電容耦合非接觸電導檢測(C4D)技術(shù)。根據(jù)本論文的研究內(nèi)容,側(cè)重論述了CE技術(shù)在食品和中藥分析中的應(yīng)用。本論文分別采用CE-AD和CE-C4D技術(shù),著重探討了水基質(zhì)中生物胺含量的測定,以及CE-AD對中草藥纈草中活性成分的分析檢測,為食品和中草藥的質(zhì)量檢驗和安全監(jiān)督提供了一種可選擇的新方法。第二章毛細管電泳-安培檢測技術(shù)在纈草生物活性成分中的檢測研究本實驗采用毛細管電泳—安培檢測法(CE-AD)同時測定了中藥纈草提取物中的八種生物藥學活性成分,八種化合物分別為：刺槐黃素、香葉木素、綠原酸、山奈酚、芹黃素、木樨草素、對羥基苯甲酸和咖啡酸。該方法通過線性范圍、重現(xiàn)性、檢出限和定量限等進行了方法確證。八種分析物的檢出限和定量限分別達到1.0x10-8～1.2x10-7g/mL和3.3×10-8～4.0×10-7g/mL。纈草樣品通過簡單的萃取程序,即可采用該方法實現(xiàn)分析和比較,所得電化學指紋圖譜可直觀顯示采自不同地區(qū)的中藥纈草及其不同纈草部位的電化學活性成分的含量差異。研究發(fā)現(xiàn),這些活性成分的含量可能會隨自然條件例如土壤、氣候、濕度等的改變而發(fā)生數(shù)量級的變化。該法己成功地將纈草中上述八種藥學活性成分進行分離和檢測,為中草藥分析提供了一種可行的方法。第三章毛細管區(qū)帶電泳-安培檢測技術(shù)在環(huán)境水生物胺檢測中的研究本實驗建立了一種基于18-冠醚-6作為緩沖液添加劑的毛細管區(qū)帶電泳-安培檢測方法,分離測定了環(huán)境水樣中的七種生物胺,包括精胺(Spm)、亞精胺(Spd)、組胺(His)、尸胺(Cad)、苯乙胺(Phe)、酪胺(Tyr)和色胺(Try)。考察了諸如緩沖液酸度和濃度、分離電壓、電極電位等實驗參數(shù)對分離檢測的影響,得到了最佳分離條件。在優(yōu)化條件下,以銅圓盤電極為工作電極,檢測電位為+650mV(vs.SCE),分離電壓為14kV,在180mmol/L18-冠醚-6/20mmol/L醋酸-醋酸鈉的緩沖溶液(pH3.6)中,七種生物胺在29min可實現(xiàn)基線分離。本方法實驗結(jié)果令人滿意,七種生物胺的含量檢出限值可達10ng/mL,峰面積(RSD≤4.8%)和遷移時間(RSD<2.4%)的重現(xiàn)性良好。該方法無需樣品預(yù)濃縮和衍生處理,為實際水樣中多種生物胺的同步定量分析提供了一個可選擇的新方法。第四章毛細管區(qū)帶電泳-非接觸電導檢測法在環(huán)境水及酒類生物胺檢測中的研究本實驗建立了一種基于18-冠醚-6作為緩沖液添加劑的毛細管區(qū)帶電泳-電容耦合非接觸電導法(CE-C4D),實現(xiàn)了對環(huán)境水樣和酒類中的八種生物胺,包括精胺(Spm)、亞精胺(Spd)、組胺(His)、腐胺(Put)尸胺(Cad)、苯乙胺(Phe)、酪胺(Tyr)和色胺(Try)的同時分析測定�？疾炝酥T如激發(fā)電壓和激發(fā)頻率、緩沖液酸度和濃度、分離電壓、進樣時間等實驗參數(shù)對分離檢測的影響,得到了最佳分離條件：激發(fā)電壓60V,激發(fā)頻率550KHz,分離電壓為16kV,緩沖溶液為150mmol/L18-冠醚-6/500mmol/L醋酸溶液。八種生物胺在24min能夠?qū)崿F(xiàn)基線分離。被測物濃度與峰面積在3-4個數(shù)量級范圍內(nèi)呈良好線性關(guān)系,最低檢測限范圍為(S/N=3)4.43×10-8-1.49x10-7g/mL。此法已被成功用于測定環(huán)境水和酒類中生物胺的分離和檢測。

    1. PrefaceCapillary electrophoresis (CE) is a new and rapidly developing separation technology which is in buffer-filled, narrow-bore capillaries, with a high-voltage power supply providing electric field; separation by electrophoresis relies on differences of characteristics of charged species. It has the characteristics of high efficiency, high speed, ultra-small sample volume, and high degree of automation. Nowadays, CE has been used to separate protein, ammo acid, inorganic ions, organic compounds, separation and determination of pharmaceutical compounds. It has been widely applied in food chemistry, medicine chemistry, analytical chemistry, clinical chemistry, environmental chemistry, and so on. In this part, the history of CE development has been retrieved, and the principle, the separation modes, the instrumental system, the technological characteristics of CE, in particular the method of capillary chromatography with amperometric detection (CE-AD) and capillary chromatography with capacitively coupled contactless conductivity detection (CE-C4D) have been introduced. According to the research area of this thesis, the application of CE in food and traditional Chinese medicine analysis has been reviewed. In this thesis,CE-AD and CE-C D has been used separately in the determination of biogenic amines in water matrix. CE has also been applied for the study on the valeriana medicinal plants. This method would find wide application in the fields of food and traditional Chinese medicine analysis.2. Study on electrochemical profiles of valeriana medicinal plants by capillary electrophoresisA method based on amperometric detection with capillary electrophoresis has been employed for the separation and determination of pharmacologically active ingredients in extracts of Valeriana medicinal plants, including acacetin, diosmetin, chlorogenic acid, kaempferol, apigenin, luteolin, p-hydroxybenzoic and caffeic acids. The method was validated for linearity, repeatability, limits of detection (LOD) and limits of quantification (LOQ), etc. The LODs and LOQs of eight compounds were found to be in the range from1.0×10-8to1.2×10-7and3.3×10-8to4.0×10-7g/mL,respectively. The proposed method was successfully applied for the analyses and comparison of bioactive components in Valerian samples after a relatively simple extraction procedure, and the resultant "electrochemical profiles" can intuitively demonstrate the content diversity of each electrochemically active ingredient in Valerian samples from different places and plant parts. It was found the content of bioactive ingredients may vary by an order of magnitude depending on natural conditions, e.g. soil, climate, humidity etc. This method has been successfully used in the analysis of pharmacologically active ingredients of Valeriana medicinal plants, which provided an alternative for the analysis of traditional Chinese medicine.3. Determination of biogenic amines in water matrix by zone electrophoresis with amperometric detectionA high-performance18-crown-6modified capillary zone electrophoresis with amperometric detection (CZE-AD)method is developed for the fast determination of seven biogenic amines (BAs), including spermine (Spm), spermidine (Spd), histamine (His), cadaverine (Cad), β-phenylethylamine (Phe), tyramine (Tyr) and tryptamine (Try). The effects of several important factors such as the acidity and concentration of running buffer, separation voltage, injection time and detection potential were investigated to acquire the optimum conditions. A copper disc electrode was used as the working electrode positioned carefully opposite the outlet of capillary at potential of+650mV (vs SCE). Under the optimum conditions, seven BAs could be well separated within29min at the separation voltage of14kV in a180mmol/L18-crown-6/20mmol/L acetate buffer solution (pH3.6). Extrapolated limits of detection for seven BAs were low to10ng/mL for the standard mixture. Repeatability of the peak current (<4.8%) and migration time (≤2.4%) of analytes for the method were achieved. The main advantages of the proposed CZE-AD method are no preconcentration and derivatization procedure, which provides an alternative method for the quantitative multi-analysis of BAs in real water matrix.4. Determination of biogenic amines in water matrix and wines by zone electrophoresis with capacitively coupled contactless conductivity detectionA high performance capillary zone electrophoresis with capacitively coupled contactless conductivity detection (CZE-C4D) method has been developed for the separation and sensitive determination of the eight biogenic amines, including Spermine (Spm), spermidine (Spd), histamine (His), putrescine(Put), cadaver (Cad), β-phenylethylamine (Phe), tyramine (Tyr) and tryptamine (Try).The effects of several important factors such as excitation voltage and frequency, the acidity and concentration of running buffer, separation voltage, and injection time were investigated to acquire the optimum conditions. Under the optimum conditions, BAs can be well separated within24min at the separation voltage of16kV in a150mmol/L18-crown-6/500mmol/L acetic acid solution and excitation voltage and frequency of60V and550KHz. Good linear relationships were established between the concentration and peak area of analytes over3-4orders of magnitude, the detection limits ranged from4.43×10-8～1.49×10-7g/mL. This proposed procedure has been successfully used for the analysis of water and wine samples.

        毛細管電泳—電化學檢測在生物胺及中草藥活性成分測定中的應(yīng)用研究

摘要6-8ABSTRACT8-10第一章 緒論12-46    第一節(jié) 毛細管電泳概述12-25    第二節(jié) 毛細管電泳電化學檢測技術(shù)25-28    第三節(jié) 毛細管電泳在食品、中草藥分析中的應(yīng)用28-38    第四節(jié) 本論文的研究目的和意義38-40    參考文獻40-46第二章 毛細管電泳-安培檢測技術(shù)在纈草生物活性成分中的檢測研究46-60    1 引言46-48    2 實驗部分48    3 結(jié)果與討論48-57    4 結(jié)論57-58    參考文獻58-60第三章 毛細管區(qū)帶電泳-安培檢測技術(shù)在環(huán)境水生物胺檢測中的研究60-72    1 引言60-61    2 實驗部分61-63    3 結(jié)果與討論63-69    4 結(jié)論69-70    參考文獻70-72第四章 毛細管區(qū)帶電泳-非接觸電導檢測法在環(huán)境水及酒類生物胺檢測中的研究72-82    1 引言72-73    2 實驗部分73-74    3 結(jié)果與討論74-80    4 結(jié)論80-81    參考文獻81-82附錄：碩士期間論文發(fā)表情況82-83致謝83

  本文關(guān)鍵詞：毛細管電泳—電化學檢測在生物胺及中草藥活性成分測定中的應(yīng)用研究，由筆耕文化傳播整理發(fā)布。