Studies On Process Quality Control Methods For The Column Chromatographic Process Of Panax Notoginseng

The column chromatography technique, which is identified as one of the most important separation techniques, has been widely used for the manufacture of Chinese medicine (CM). Currently, the CM column chromatographic manufacturing is still not able to ensure the batch-to-bath reproducibility of the products due to lacking advanced process quality control methods. For this reason, taking the column chromatographic process of Panax notoginseng as the research carrier, studies focusing on the process quality control methods for CM column chromatographic process were carried out. The main innovation and academic contributions of this dissertation are summarized as follows:1. The ultraviolet (UV) spectra combined with multivariate statistical process control (MSPC) method were used to in-line monitor Panax notoginseng column chromatographic process via score, Hotelling T2 and DModX control charts. This method could be used to evaluate the batch-to-bath reproducibility and detect process variations.2. The elution amount of total saponins and apparent purity of total saponins in effluent were used to represent the performance of elution process. In order to achieve acceptable elution performance, multivariate regression models, which mapped the influence of the total saponins concentration in loading solution, elution time to the elution performance, were established. The potential operating range of the total saponins concentration in loading solution and elution time were determined under the framework of quality by design (QbD).3. The total saponins adsorption amount and loss rate were used to represent the performance of adsorption process. Multivariate regression models, which mapped the influence of the total saponins concentration in loading solution, adsorption time to the adsorption performance, were established. Based on the total saponins concentration in loading solution, the adsorption time could be adjusted early-on for acceptable adsorption performance. The UV spectra of effluent during adsorption process in different batches were collected. Principal component analysis (PCA) was carried to visualize the UV spectra and the principal components were obtained. Multivariate regression models, which mapped the influence of the principal components of the UV spectra, elution time to the elution performance, were established. Based on the principal components of the UV spectra of adsorption process, the elution time could be adjusted early-on for acceptable elution performance. These studies provided a new idea for the process quality control of CM column chromatographic process.4. The experimental breakthrough curves of notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1 and ginsenoside Rd were obtained via an offline ultra performance liquid chromatography (UPLC) method. The Yoon-Nelson model was used to fit the breakthrough curves of five saponins in different batches and the model parameters were obtained. The relationships between saponins concentrations in loading solution, flow rate and the model parameters were investigated by multivariate regression models. According to the multivariate regression models, the model parameters were calculated and the breakthrough curves of five saponins could be predicted. The predicted breakthrough curves of five saponins provided guidance for process quality control of Panax notoginseng column chromatographic adsorption process.5. In-line UV spectra combined with moving block standard deviation (MBSD), difference between the moving block average and the target spectrum (DMBA-TS), soft independent modeling of class analogy (SIMCA) and partial least squares discriminant analysis (PLS-DA) were used to determine the end-point of adsorption process. Compared with MBSD and DMBA-TS, the SIMCA and PLS-DA were more suitable for the determination of the adsorption end-point. The uninformative variable elimination-partial least squares (UVE-PLS) models based on spectroscopy were established for quantitative analysis of the saponins concentrations in effluent during adsorption and elution process. These researches provided a theoretical basis for real-time process quality control of Panax notoginseng column chromatographic process.