Application Of Capillay Electrophoresis In Several Antihistamines

Capillary electrophoresis(CE), also called High Performance Capillary Electrophoresis (HPCE), is a new analytical technique considered as one of the powerful analytical Chemistry branches, and also an important improvement of the separation sciences after high Performance liquid chromatography in the early1980s.Laser-induced fluorescence (LIF) detector is one of the most sensitive methods, applied in CE due to its low LIF slow background and extremely small sample volume. The application of LIF with CE becomes more and more widely. The application fields include environmental and clinical analysis, peptide and protein analysis.At the thesis, we mainly studied the contents of two parts: 1.By using of Capillary electrophoresis, we studied several antihistamine compounds chiral separation. There are chlorpheniramine, carbinoxamine, dioxopromethazine, doxylamine, oxomemazine, hydroxyzine hydrochloride, cetirizine. We investigated the impact of separation of several factors:(1)Chiral selector type of the impact for separation: We investigated the impact ofβ-CD,γ-CD, CM-β-CD, HP-β-CD, Tri-OMe-β-CD, DM-β-CD on chiral separation of antihistamines . Results: CM-β-CD as chiral selector were more effective on chiral separation the five antihistamines.(2)Chiral selector concentration of the impact on separation: Chiral selector concentration is also affected enantioseparation an important factor. We investigated CM-β-CD of 5～15mmol/L about the affect on chiral antihistamine drugs separation. Results: CM-β-CD concentration range 5～15mmol/L, with the concentration increasing, the separation was better. But.the separation time is longer. Considerated the separation and analysis time, we selected the the best separation concentration 10 mmol/L.(3)Running buffer of pH value on the impact of separation: We investigated the buffer solution pH value of 3.0, 3.5, 4.0, 4.8, 5.5 , 6.3on separation antihistamines of the effects. Result: We selected 3.5 as the best pH value of chlorpheniramine and oxomemazine.We selected 4.8 as the best pH value of carbinoxamine, dioxopromethazine and doxylamine.(4)The voltage on the impact of separation: We studied different voltage on the impact of separationthe in this experiment. Result: The voltage have an impact on the separation and analysis time.We considerated the separation and analysis time, then we selected the best voltage: Chlorpheniramine 20kV, Carbinoxamine 20kV, Oxomemazine 12kV, Doxylamine 16kV, Dioxopromethazine 12kV.(5)Column temperature on the impact of separation: In this experiment we investigated the column temperature on the impact of separation at the same time.The Column temperature not only affects the migration time, but also affected the cyclodextrin with enantiomers of inclusion complexation between the balances. Considerated,we selected the column temperature: Chlorpheniramine 25℃, Carbinoxamine 25℃,Oxomemazine 16℃,oxylamine 16℃,Dioxopromethazine 16℃.2. The sotalol was derivatized by the fluorescein isothiocyanate (FITC), and then analyzed the derivatization by capillary electrophoresis. We used the detect method of capillary electrophoresis-laser induced fluorescence. Investigated the derivatization of buffer solution(50mmol/L borax solution, 0.01 mmol/L sodium hydroxide solution, 200mmol/L carbonic salt solution, 200mmol/L phosphate solution) on the impact in this reaction. Results: Results is better derivatization in the borax solution, interference at least, the strongest signal can clearly be saw the product of derivatives of the peak. However, in other three buffers after derivatization of the product have more interference.So we selected 50mmol/L borax solution.We also investigated the derivatives derivatization reaction time within the 2～24h on the impact of rection. Investigated the derivatization of temperature and capillary electrophoresis separation conditions (as for: the buffer solution concentration, the buffer of pH value, temperature and voltage and so on). So we can select the most optimized derivatization conditions and separation conditions. Results: We ultimately selected the conditions :50mmol/L borax solution as the derivative of the solution , reaction time 12h, at room temperature, holding the dark. the conditions of electrophoretic separation is 25mmol / L of borax solution (pH = 9.2) as separation buffer , separation voltage of 20kV, separation temperature 25℃, the effective length of capillary column 37cm.