Development Separation And Determination Of Ten Key Biogenic Amines By Capillary Electrophoresis

Biogenic amines (BAs) are existence not only in the organization of animals and plants,butalso in all kinds of food, especially in the fermentation products. BAs plays an important role onhuman respiratory system, and has the inseparable relations to circulatory system, immune system,and even the nervous system.Some of BAs are important markers of clinical detections of diseasessuch as Parkinson’s disease, pheochromocytoma. But excessive intake can also cause poisoning andallergic reaction, so the detection of BAs is one of the important indicators of monitoring thedetermination of the food freshness and safety. Compared with the traditional technique of the highperformance liquid chromatography (HPLC), capillary electrophoresis (CE) has characteristics,such as more separation modes, higher separation efficiency, faster analysis, less sampleconsumption, etc, it has been widely used in the separation and analysis of inorganic ions, sugars,amino acids, proteins, nucleic acids, environmental samples, drugs, food, and chiral enantiomers.Many BAs often do not have the UV absorption. Traditional UV and fluorescence detection (FD)often take the derivative method to analyze the BAs, but operation is too complex, in this paper, CEtechnology without derivatization, analysis BAs at the same time with direct or indirect UVdetection, has been applied to the analysis of the actual sample such as urine, soy sauce, etc.First chapter briefly introduced the related theories and separation modes of CE, especiallytheir separation analysis applications to BAs, this paper introduced the properties of BAs and itsrole in the human body and food, comparesd the different approaches to the analysis of BAs, suchas thin layer chromatography (TLC), gas chromatography (GC), liquid chromatography (LC) andCE, etc.This paper summarizes CE technology commonly used detectors and its application in BAs analysis, finally puts forward this topic conception.Second chapter choosed10BAs with UV absorption (phenethylamine (PEA), histamine (HA)and tryptamine (Try), tyramine (TA), serotonin (5-HT), octopamine (OA), dopamine (DA),norepinephrine (NE), epinephrine (E), and carnosine (CAR)), developed their CE-direct UVdetection analysis and separation method, and under the standard sample optimized the separationvoltage, detection wavelength, the constitute of buffer, the concentration of buffer, the pH of thesystem, and the effect of different additives and their concentration, and finally found the besteffect in150.0mmol L-1sodium phosphate buffer system (pH=6.10),, containing the1.0mmol L-1borax,, separation voltage at+10kV, detection wavelength at200nm (UV). The method cananalyse BAs without derivatization,10kinds of analytes LODs between0.2to1.3μmol L-1, theRSD of separation time and peak area in0.08-0.12%and2.85-4.51%, respectively.Third chapter explored the separation method of four BAs without UV absorption (cadaverine(Cad), putrescine (Put), spermine (Spm) and spermidine (Spd)) using indirect UV detector.Discusse the selection of indirect UV detection electrolyte and its concentration, the effects of thepH of the buffer system to separation, but so far we can not make all4kinds of BAs separation, soconsider adding different kinds of additives to achieve, and find EDTA raised the separating degreeof four BAs, made its baseline separation, finally summed up the optimum separation conditions of10mmol L-1imidazole-acetic acid buffer system (pH=4.50), containing0.5mmol L-1EDTA,separation voltage at+20kV, detection wavelength at200nm.Fourth chapter put forward the direct and indirect UV detection at the same time, used theselection of14BAs (PEA, HA Try, TA,5-HT, OA, DA, NE, E, CAR, Cad, Put, Spm and Spd) asthe standard sample, preliminary explored the the possibility of this method to analyize two kindsof BAs at the same time (UV absorption and no UV absorption),and the influence of the backgroundelectrolyte concentration, pH, additives and their concentrations to the separation condition of14kinds of BAs, and the optimal conditions to make the selection of12kinds of BAs.Fifth chapter firstly used the method developed in this paper, determinated of several BAs ofathletes’ urine, contrasts their changes of contents before and after the athletes fatigue, and exploredthe relationship between the changes and sports fatigue. And then we analysed contents of Cad, Put,Spm and Spd of soy sauce by CE-indirect UV detection. Finally because in the detection processof actual samples such as urine, soy sauce and so on, the analysis process tended to be affected bythe sample’s complex matrix, therefore in order to eliminate the interference, we need to developthe pre-treatment technology of actual samples, this paper only used the solid phase extractionmethod and carried on the preliminary exploration, used Cad, Put, Spm and Spd as standard sample,CE as detecting tool, explored the adsorption behavior of BAs on different solid phase extractioncolumn (HLB, C18, MCX), hoped the method has a better application in the future.