Quality Analysis On Key Processes Of Glucosamine Production By Using Miniature Near Infrared Spectrometer

Glucosamine(GlcN)is a popular functional product of bone-protecting,and has a great market potential.The main production methods of it containing hydrolyzing the shells of shellfish,chitin enzymolysis and microbial fermentation,and more and more manufacturers turn to use the last approach to get GlcN which is free from the supply and the price of raw materials and environmental restrictions.While the zymotechnics offers many advantages,it costs a lot on analysis due to it is a nonlinear dynamic process with high complexity and multiple parameters,and the time-consuming of common chemical determination methods prevents timely feedback of information or cost reduction,which affect the quality of the products and squeeze on profit.Therefore,it will make a significant sense on reduce the production cost and improve the quality of products if introduced an advanced Process Analytical Technological(PAT)into the production process of GlcN to achieve the quick detection of important quality indices in the key processes with a PAT tool characterized by fast,accurate,green and low-cost-Near Infrared Spectroscopy(NIRS).NIRS has been a relatively mature and widely used PAT tools after more than half a century development,and a wide range of the near infrared spectrometers are available in present.With the development of science and technology,the design of near infrared spectrometers are under the trend of miniaturization,portability and cost minimization,and it will be used more and more in the pharmaceutical industry.This research will be analyze feasibility of NIRS applied in determination the concentration of GlcNAc at first because it is the direct product of fermentation process,and it will be GlcN thr-ough deacetylation on subsequent process.The next,application feasibility of NIRS in the production process of GlcN will be investigated by introduced it into the fermentation process and concentration process.The specific research contents are listed as below:(1)The feasibility of NIRS applied in the determination of GlcNAc under given systemThis research analyzed the feasibilities of using Fourier transform near infrared spectrometer(FT-NIR)and miniature near infrared spectrometer to determine the concentration of GlcNAc under given system,and conducted comparative analysis between the two instruments prediction results,which lay a theory foundation for introducing the miniature near infrared spectrometer into the production process of GlcN.At first,pure water-GlcNAc system was taken as research object and 40 samples prepared in laboratory with concentration range from 4 g/L to 160g/L.The spectra collection of all samples were accomplished by a FT-NIR and Micro-NIR1700.Before established the Least Square Regression(PLSR)quantitative analysis model for determining the concentration of GlcNAc,content-grads method was used to split the sample set into two parts:27 samples as calibration set and 13 samples as validation set and pretreatment was used to improve spectral quality.Finally,Passing-Bablok regression was selected to compare the prediction accuracy between the two instruments,and result showed there was no significant difference.Next,adopted the same method to analysis the feasibility of NIRS,especially the miniature near infrared spectrometer,applying in determining the concentration of GlcNAc under fermentation broth complex system,and the results proved the possibility of it.In addition,the result of Passing-Bablok regression showed there was no significant difference between the two instruments even in the complex system.(2)The determination of GlcNAc concentration and optical density(OD)values in the fermentation process by using miniature near infrared spectrometerThis research introduced Micro-NIR1700 near infrared spectrometer into the fermentation process to explore its feasibility to determine the concentration of GlcNAc and OD values in fermentation liquor.Firstly,135 samples were collected on the normal production lines.The concentrations of GlcNAc were measured by High Performance Liquid Chromatography(HPLC)and the OD values were determined by spectrophotometric method.Then,the PLSR quantitative model of GlcNAc concentration and OD values were both established by 90 samples and validated by 45 samples,and them would be further optimized by comparing different dataset partition,pretreatment and variable selection methods.The best models of GlcNAc concentration and OD values got the values ofRc2?Rp2?RMSECV and RMSEP like 0.994?0.995?2.7927 g/L and 1.9468 g/L and 0.957?0.947?7.8032 and 7.1084,respectively.The research results showed that the established models have the abilities in quick determining the GlcNAc concentration and OD values with fine prediction results.(3)The application research of NIRS used in the determination of GlcNAc content in the concentration processThis study applied Micro-NIR1700 near infrared spectroscopy into the concentration process in order to achieve the quick determination of GlcNAc content and the endpoint.First,we simulated two batches of concentration process and got 37 samples,then sampled 13 from manufacturing process.In total,50 samples were obtained and collected spectra using Micro-NIR1700 near infrared spectroscopy.Before established the best quantitative model of the content of GlcNAc,variable dataset partition methods,spectra pretreatment methods and variable selection methods were carried out and compared.Finally.38 variables were selected to established the optimal model,and its values of Rc2?Rp2?RMSECV and RMSEP were 0.992?0.991?1.3436%and 0.9496%,respectively.The results showed that the established model has good prediction ability and has capacity to determine the content of GlcNAc and the end point quickly.The innovations of this research are listed as below:(1)Compared the accuracy of FT-NIR and Micro-NIR spectrometers on determining the concentration of GlcNAc under fermentation background for the first time,and proved the feasibility of Micro-NIR applied in the production process of GlcN.(2)This study introduced Micro-NIR spectrometer into the fermentation process of GlcN for the first time,and proved the rapid and simultaneous determination of GlcNAc concentration and OD values was feasible.(3)This study introduced Micro-NIR spectrometer into the concentarion process of GlcN for the first time,and achieved the quick determination of GlcNAc and end point in this process.