Study On The Rapid Determination Of Reduced Glutathione Tablets With FT-NIRs And Chemometrics

The clinical applications of reduced glutathione tablets are mainly used for the treatment of liver damage, antivirus and antiparkinsonism. Currently, the analytical methods for reduced glutathione tablets include HPLC, UV and so on. However, these methods have disadvantages, such as complex sample preparation, destruction of samples, time consumption and pollution.Diffuse reflectance Fourier transform near infrared spectroscopy (diffuse reflectance FT-NIR) combined with chemometrics can be used for the determination of solid samples fast and non-destructively. On the other hand, the method can be used for the process analysis.In this study, the methods for the rapid determination of reduced glutathione tablets were established based on diffuse reflectance FT-NIR combined with chemometrics and the reference values (HPLC) for quality control and process analysis, respectively. Root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP) and correlation coefficient(R) were used for the model evaluation.OBJECTIVES:1. To establish a method for the determination of reduced glutathione tablets with FT-NIR diffuse reflectance spectra obtained by an integrating sphere attachment for quality control.2. To establish a method for the determination of reduced glutathione tablets with FT-NIR diffuse reflectance spectra obtained by a fiber optic probe for process analysis.METHODS:1. Measurement of FT-NIR spectra of reduced glutathione tablets(1) Measurement of FT-NIR diffuse reflectance spectra by an integrating sphere attachmentThe spectra of reduced glutathione tablets were measured by FT-NIR analyzer with an integrating sphere attachment. The measurement conditions were: scan region: 10 000 4000cm^-1, number of scans: 64, resolution: 8cm?1, ambient temperature: (22±1)℃, ambient humidity: (55±5 )% RH.(2) Measurement of FT-NIR diffuse reflectance spectra by a fiber optic probeThe spectra of reduced glutathione tablets were measured by FT-NIR analyzer with a fiber optic probe. The measurement conditions were: scan region: 10 000 4000cm^-1, number of scans: 32, resolution: 8cm?1, ambient temperature: (22±1)℃, ambient humidity: (55±5 )% RH.2. Determination of the content (HPLC) of reduced glutathione tablets The reference values of reduced glutathione tablets were measured by HPLC analyzer. The chromatographic conditions were: column: C₁₈ (4.6 mm×250 mm, 5μm), mobile phase: 0.1% trifluoroacetic acid-acetonitrile (98:2), column temperature: 30℃, flow rate: 1ml/min, detection wavelength: 210nm, injection volume: 20μl.3. Calibration and validation of the models for the determination of reduced glutathione tabletsSpectral ranges were selected. The spectra processing were performed respectively for quality control and process analysis. Then, PLS and BP-ANN models were established respectively for quality control and process analysis. RMSECV, RMSEP and R were used for the validation of the models.RESULTS:1. Methods for the determination of reduced glutathione tablets for quality control(1) Calibration and validation of PLS model: The PLS model was established with 5 factors. RMSECV was 0.02413, RMSEP was 0.00857 and R for the prediction set was 0.97254.(2) Calibration and validation of BP-ANN model: The structure parameters of the BP-ANN were (11,5,1). RMSECV was 0.02673, RMSEP was 0.02808 and R for the prediction set was 0.63906.2. Methods for the determination of reduced glutathione tablets for process analysis(1) Calibration and validation of PLS model: The PLS model was established with 7 factors. RMSECV was 0.03403, RMSEP was 0.0100and R for the prediction set was 0.93311.(2) Calibration and validation of BP-ANN model: The structure parameters of the BP-ANN were (8,4,1). RMSECV was 0.04043, RMSEP was 0.02123 and R for the prediction set was 0.80131.CONCLUSIONS:1. The PLS model built with the diffuse reflectance spectra obtained by an integrating sphere attachment could be used for the rapid prediction of the content of reduced glutathione tablets for quality control.2. The PLS model built with the diffuse reflectance spectra obtained by a fiber optic probe could be used for the rapid prediction of the content of reduced glutathione tablets for process analysis.