Study On Chiral Separation And Determination Of Phenylalanine Enantiomer By Cyclodextrin-capillary Zone Electrophoresis

Phenylalanine(Phe) owns DL-,D- and L- configurations, L-Phe of which is one of eight kinds of necessary amino acids that can not be synthesized by the human being. It is a fundamental component of clinical physician and pharmacist, and is the important constituent of food additive.And D-Phe is an important intermediate material of aids virucides.So the separation of phenylalanine enantiomers is necessary .Then their chiral analysis is the first question to solve.As yet, simple and fast chiral analysis method of phenylalanine has not been reported. This dissertation mainly deals with the chiral analysis of phenylalanine.Firstly, a method whose chiral selector is commercial composite 2,6-dimethyla-ted-β-cyclodextrin(DM-β-CD) to determine the contents of D,L-phenylalanine is established. Phenylalanine can be determined directly with no derivative, because they have strong absorption in the UV range. The influential factor such as the type of buffer, pH, the concentration of phosphate salt, the concentration and type of chiral selector, temperature, voltage, the size and material of capillary, etc, are studied. Finally, the optimal chromatographic parameters are decided as follows: the material and size of capillary: 75μm×40㎝fused silica uncoated capillary(effec-tive length is 30㎝)detective wavelength: 205nmvoltage: 22kvelectrode: normaltemperature: capillary temperature is25℃,sample temperature is 25℃buffer: 40mmol·L-1 sodium dihydrogen phosphate with 60mmol·L-1 DM-β-CD, pH=2.2inject mode: 0.5psi×5sec,followed by 0.1psi×10sec inject of distillation water and the rinse outlet is buffer mode of rinse between two run: capillary rinsed by distillation water for 1min,rinsed by 0.5mmol.L-1KOH for 2min, rinsed by distillation water for 1min,rinsed by 1%H3PO4 for 1 min, rinsed by distillation water for 1min, rinsed by buffer for 1min in turnOn these conditions, the linear ranges of D-Phe and L-Phe are 0.00338-0.10813 g·L-1(R=0.9995)and 0.00354-0.11327 g·L-1 (R=0.9996);the ranges of recycle rate are 97.8%~ 101.1% and 98.1%~102.4%; the relative standard deviations expressed by RSD are 3.55% and 3.82%; the detectable qualities are 0.00162 g·L-1and 0.00161 g·L-1;the minimum determinable qualities are 0.00323 g·L-1and 0.00322 g·L-1.The method is simple, quick and accurate, it can be finished in twelve minutes and D,L-phenylalanine can get a baseline separation . This simple and fast analysis method will play an important part in the produce and quality control of phenylalanineThis thesis also make a research on the synthesization of partially methylated β-cyclodextrin which will decrease the analysis cost and we then take a preliminary identification of it. The result of chiral separation and determination of D,L-phenylal-alanine with self made partially methylatedβ-cyclodextrin to the is good. The chromatographic parameters with self made partially methylatedβ-cyclodextrin are as follows:buffer: 40mmol·L-1 sodium dihydrogen phosphate with 80mmol·L-1 self made partially methylatedβ-cyclodextrin,pH=1.85voltage: 20kvother parameters: the same as the chromatographic parameters with commercial composite DM-β-CD...