Synthesis Of Oseltamivir Molecular Imprinted Polymers And Its Evaluation

Molecularly imprinted polymers (MIP) have many advantages, such as strongly molecularrecognition, simple and rapid preparation of stationary phase, relatively stable, lower solventconsumption, template and MIP can be recycled, etc. It shows a good prospect in the enantiomers andisomersseparation, solid phase extraction, controlled release drug delivery systems, clinical druganalysis, membrane separation technology, mimic enzyme catalysis, chemical sensors and other areas ofbionic. The method of traditional antiviral drugs screening has some shortcomings, which containscomplicated process, higher risk and rigorously experimental conditions. To establish non-biologicaldrug screening technology of anti-viral active component screening, separation and identification fromChinese traditional herbs through MIP will have important application value. In this paper, oseltamivir(OS) was used as template to prepare molecularly imprinted polymers (MIP) by bulk polymerization,which had been evaluated. We hope using OSMIP-HPLC-MS method to Screen of new anti-influenzavirus compounds.1. Anti-influenza drugs and their pharmacological action, the definition of non-biological drugscreening methods and their application, the synthesis of molecular imprinted polymers and its reagentselection, and the application of MIT in the separation of the active ingredient group, screening activeingredients, and the field of antiviral drugs was reviewed.2. Thin layer chromatography was used for elementary identification of OS. The developingsolvent was 2 mL of methanol, 4 mL of ethylacetate, 2 mL of toluene and 1 mL of triethylamine. Glassplates, which pre-coated with silica gel GF254, were detected with UV light of 254 nm. The resultshowed that the presence of OS was indicated by a dark spot at a travel distance of about 0.5.3. The further identification and quantitation of OS were tested by mass spectrometry, tandem massspectrometry (MS/MS) and multiple reaction monitor (MRM). Methanol-water (75 : 25, V/V) and C₁₈column were used as mobile phase and stationary phase, respectively. The ion monitored was OS [M +H]+ m/z 313.30, and product ion 166.20 was used as the quantifying ion whereas ion 208.20 was used asthe accessorial qualitative ion. And there was a good correlation coefficient between 5^-160 ng·mL^-1 withMRM method,the correlation coefficient of linearity R² was 0.9995, the average recovery was 100.94%,the intra- and inter- day precision values were 99.89% and 98.90% , respectively.4. To optimize the process parameters of OS-MIP, U₈(8₂×4₂) uniform design table was made bymeans of sorption quantity. The optimized parameters were 0.1 mmol OS, functional monomer 0.69mmol, EGDMA 5 mmol, 30% AA + 70% 4-VP as functional monomer, toluene as porogenic solvent.The sorption capacity of prepared OS-MIP for OS was 220.9543μg·50 mg–1, whereas the prepared NIPfor OS was 12.0817μg·50 mg^-1.5. To investigate the difference of OS-MIP from NIP, scanning electron microscopy was employedto observe their structure. The SEM image clearly showed that pores were embedded in the OS-MIP andNIP networks, and that there were substantial differences in morphology between OS-MIP and NIP. NIP had a smoother structure with fewer and smaller cavities than were found on OS-MIP.6. In order to evaluate affinity and selectivity of OS-MIP for template, OSMIP-HPLC-MS methodwas used to detect the retention time of OS, aspirin, chlorogenic acid, forsythin and the complexmatrix on OSMIP-HPLC column. TIC showed the column can separate the template molecule fromother compounds. The retention time of its analogs was about 35min, but the template molecule wasabout 110 min (due to the different physical and chemical properties, the retention time will change).In conclusion, thin layer chromatography method was simple, fast and can be used to identify theauthenticity of OS in the market. The further identification and quantitation of OS were tested by liquidchromatography tandem mass spectrometry, which provided technical support for further detection ofmolecularly imprinted polymers sorption capacity for OS. Uniform design was used to optimizeOS-MIP synthesis parameters. The prepared OS-MIP has a stronger sorption for OS. Scanning electronmicroscopy was employed to observe the polymers'structure. The results showed the preparedOS-MIP had consistent structure with imprinted polymer morphology. The experimental results ofevaluating OSMIP-HPLC column showed the self-made columns had specific affinity and selectivityfor OS to separate the template molecule from other compounds. The results showed that theOSMIP-HPLC-MS method could separate OS from matrix, and it may also be helpful in finding moreactive analogs of OS from Chinese traditional herbs.