New Cyclodextrin Chiral Stationary Phase On Fluoxetine Enantiomers Split

The production of individual enantiomers has become more and more important as the increased knowledge of the different medicine effect of enantiomers. There are several approaches to obtain enantiomerically pure chemicals. They are asymmetric synthesis, chemical derivation, crystallization, biotransformation, inclusion, membrane separation and chiral chromatography. The chiral chromatography is more attractive and has been developed rapidly in recent years. However, research was concentrated on the chiral analysis and chiral recognition mechanism, and the study on the preparative separation is not excessive. This work chooses the chiral widely used antidepressant drug — fluoxetine as the object to study the common technique of chiral chromatographic separation. After comparing the chiral resolution methods of fluoxetine, we intend to separate the enantiomers of fluoxetine by the perphenylcarbamoylated B-cyclodextrin bonded CSP chromatography.The experimentation of dissertation is consisted of four parts: the preparation of perphenylcarbamoylated B-cyclodextrin bonded chiral column, the static adsorptive capacity of CSP, the chromatographic separation of fluoxetine enantiomers on self-made column and the analysis of the chromatographic process.In the first part, the perphenylcarbamoylated B-cyclodextrin CSP was prepared step by step. In the project, mono-6-p-toluenesulfonyl-B-cyclodextrin, mono-6-N-allylamino-β-cyclodextrin and mono-6-N-allylamino-perphenylcarbamoylated-β-cyclodextrin were synthesized and purified;and then, the last was bonded to the y-aminopropyl silica gel. The slurry method was applied to prepare the HPLC column with methanol as the packing solvent. After suspended in CCl₄-dioxane (2/1, V/V) and sonicated for 10 min, slurry of CSPs were packed into a stainless steel column at a pressure of 28 MPa. At 0.5 mL/min of flow rate, the theoretical plate numbers of two columns were 6222 and 5630, respectively.Secondly, separation of fluoxetine enantiomers on five chiral stationary phases (Chiralcel OD-H, Chiralcel OJ-H, Chiralpak AD-H, Cyclobond I 2000 DM and Kromasil CHI-TBB) was investigated. Baseline separation was obtained on OD-H, AD-H, and Cyclobond I 2000 DM while the best was obtained on the last one. Afterthe analytical method was determined, the static adsorptive capacity of CSP was tested. Experimental results demonstrated that the chiral selector has selectivity, that is, the adsorption of S-fluoxetine on perphenylcarbamoylated B-cyclodextrin is easily.The enantiomers of fluoxetine were separated on perphenylcarbamoylated B-cyclodextrin bonded CSP with the mobile phase of methanol/TEAA. After determining of the chromatographic model, the composition, ionic strength and pH of mobile phase were investigated, as well as the flow rate and column temperature. The first eluate by self-made column is the R-enantiomer. Besides, paroxetine, naproxen, propranolol and alprenolol were baseline separated on the column;the preparation of CSPs and packing of columns are repeatable.At the end of the dissertation, the chromatography separation process of fluoxetine enantiomers was studied. The effects of the composition and flow rate of mobile phase on the throughput and energy of solvent evaporation were studied. It was found that the throughput increases with the increase of the content of methanol and the flow rate of the mobile phase. The energy of solvent evaporation increases with the increase of the flow rate of mobile phase, but decreases with the increase of the content of methanol in mobile phase. Taking into account of the resolution, throughput and energy of solvent evaporation, the compromise composition of mobile phase may be methanol/TEAA=40/60 (V/V- The elution curve was analyzed with moment analysis. The equilibrium constant of solute between the mobile and stationary phases can be calculated from the first absolute moment, the total mass transport resistance can be calculated from the second central moment. A linear driving force model was set up to simulate the experimental elution curve. Based on the model, effects of loading and flow rate of mobile phase on separation process were studied. The results could be used to scale up of preparative chromatography.In this paper, a new CSP perphenylcarbamoylated 6-cyclodextrin bonded chiral stationary phase was prepared. And it was the foundation of preparation of some enantiomerically pure chemicals, such as fluoxetine enantiomers etc.