The Research On Separation And Detection Of Amlodipine Besylate

A large amount of researches and clinical practice indicated that the enantiomers of chiral drugs always have significantly differences in pharmacology activity, metabolic process and toxicity performance in human bodies. With the rising morbidity and mortality of angiocardiopathy, the demand for cardiovascular drugs is increasing continuously. Many of those cardiovascular drugs have one or more chiral atoms, and it could probably bring serious side effects and the corresponding syndrome when provided with racemic drug. At present, most of the common methods for separating the chiral drug are small-scaled in the laboratory and hard to realize large-scaled production. Therefore, it is a significant work to construct a relatively useful method for the separation of cardiovascular chiral drugs with large-scale. In this thesis, a chiral cardiovascular drugs-amlodipine besylate was chosen as the research object. Combined with a variety of detection methods such as HPLC and Fluorescence Spectroscopy, the controlled molecular imprinting technique/hollow fiber membrane and aqueous two-phase extraction technology were used to carry out the research on separation and detection of amlodipine besylate. This research of this thesis aims to achieve the separation of chiral drugs amlodipine besylate with high throughput, so as to provide technical support for the industrial production of cardiovascular chiral drug about amlodipine besylate. The details are as follows:1. Using S-amlodipine besylate as template, molecularly imprinted with PVDF membrane as supporting membranes, a high-capacity imprinted-membrane materials were prepared. The optimum conditions for the synthesis of polymer membranes were ivestigated. The imprinted layer of molecularly imprinted membrane had memorial hole to S-amlodipine besylate, so it had high selective property and the maximum adsorption factor was1.52. Compared with non-imprinted membrane and blank membrane, the adsorption process for S-alodopine besylate was nonspecific adsorption. SEM result indicated that the membrane surface was coated with a layer of dense imprinted polymer.2. After the synthetic surface controllable and high-capacity imprinted-membrane materials of S-amlodipine besylate were assembled to the independently developed membrane modules, the study was carried out to separate the chiral drug of amlodipine besylate with high-throughput, in order to achieve large-scale separation. Then through the study on amlodipine besylate by high-throughput screening, the scale of separation was achieved. Also, the mathematical model of the hollow fiber membranes to separated amlodipine besylate: S/R=0.780e0.096NTU was established. Using the hollow fiber membranes with the length of175cm, the purity of amlodipine enantiomers could be up to90%. Some new progresses were expeated to appear in the high separation of chiral drugs, which would provide the technical basis for the industrial production of cardiovasclular chiral drug about amlodipine besylate.3. Based on the Polyethylene glycol(PEG)/Ammonium sulfate aqueous two phase system with sulfobutyl-β-cyclodextrin (SBE-β-CD), HPLC was proposed for the chiral separation of amlodipine. The effect of the molecular weight of PEG, the amount of SBE-β-CD and PEG, pH, temperature, the amount of ethanol were investigated. The results showed that all the above factors had serious effects on the separation result. SBE-β-CD was inclined to recognize S-amlodipine besylate. PEG6000was finally chosen as the study object in this experiment. Under the optimum conditions, the separation factor could be0.72. This method has explored a new path for preparative separation of chiral drug of amlodipine besylate.4. The separation of amlodipine besylate was investigated by using the Polyethylene glycol(PEG)-salt aqueous two pHase system with hydroxypropyl-β-cyclodextrin (HP-β-CD). The effect of the molecular weight of PEG, the amount of PEG, the amount of HP-β-CD, pH and temperature were investigated. The result showed that HP-β-CD was firstly inclined to recognize S-amlodipine besylate. The optimal experimental conditions were0.25mmol/L HP-β-CD,2.0g ammonium sulfate,3.0g PEG6000and pH5.0. Under the optimum conditions, the separation factor was0.74.5. Based on the agility and practicability of chiral mobile phase additives method, a simple, rapid and sensitive method was developed for the separation of amlodipine besylate in16min, which used sulfobutyl-β-cyclodextrin(SBE-β-CD) and Polyethylene glycol (PEG-20M) as dual additives. At the same time, the mechanism of the separation was discussed and the result was satisfactory. This method was more rapid and easy to operate compared with other separation methods. Also, it provided a more convenient method for the detection of S-and R-amlodipine besylate.6. To set up a rapid, simple fluorescence spectrometry for the determination of the S-amlodipine besylate. The spectral properties between amlodipine besylate and SBE-β-CD has been evaluated and the interaction mechanism has been investigated. Based the fluorescence enhanced effect of SBE-β-CD, the enhancement of the relative fluorescence intensity was observed on S-amlodipine besylate at455nm. Under the optimum conditions, the calibration curve exhibited an excellent linear relationship with a correlation coefficient of0.9998. The limits of detection was0.17μg/mL and the limits of quantification was0.55μg/mL. The method was shown to have the advantage of simple, low cost and wide detection range.