Identification of bear bile powder and its adulterants by using DNA barcoding technology / 中国中药杂志

To identify the precious bile powder and its adulterants by DNA barcoding, and establish its standard experimental process to ensure the safe and effective utilization. Total twelve sequences from samples of bear bile powder which come from Ursus thibetanus for DNA extraction, PCR(polymerase chain reaction) and sequence, then using CodonCode Aligner V 7.0.1 shear primer region to obtain COI sequence. The COI sequences of U. arctos and their adulterants were obtained from GenBank. MEGA7.0 software was applied for analyzing mutation, calculating intraspecific and interspecific K2P(Kimura 2-Parameter) genetic distance and constructing the Neighbor-joining tree(NJ). The results showed that the maximum K2P genetic distance of bear bile powder of U. thibetanus and U. arctos are far less than minimum K2P genetic distance within its adulterants species, and the results of NJ tree demonstrated that each species could be distinguished from the counterfeits obviously. DNA barcoding is a safe, convenient and reliable technique for species identification, and it is important to establish the standard sequence of COI sequences for animal medicines.

Primary exploration of key influential factors recognization method of Tanreqing injection / 中国中药杂志

To recognize the key influential factors during the liquid preparation process of Tanreqing injection, the near infrared(NIR) spectra of the raw materials and the operating parameters of 24 batches of physical manufacturing were recorded as independent variables, and the total soluble solids contents and the light inspection acceptance rate of the final products were collected as dependent variables. The calibration models were developed using the partial least-square regression (PLSR) method, and the correlation coefficients between the independent variables and the dependent variables were calculated. For the quantitative models, the correlation coefficients for the calibration and inner cross validation of total soluble solids contents and the light inspection acceptance rate reached 0.911 9, 0.724 2 and 0.873 8, 0.795 9, respectively. Using the correlation coefficients diagrams, several key influence factors were preliminarily determined, and the physical significance were analyzed combined with production experience. This work demonstrated that NIR spectroscopy with PLSR algorithm could be used for the key influential factors recognization during the liquid preparation process of Tanreqing injection and can be popularized to solve similar problems..

Screening and identification of multiple components in Tanreqing injection using RP-HPLC combined with DAD and ESI-TOF/MS / 中国天然药物

A liquid chromatography coupled with diode array detector (DAD) and electrospray ionization time-of-flight mass spectrometry (ESI-TOF/MS) method was developed for the screening and identification of the multiple components in Tanreqing injection, a well-known Chinese medicine injection in China. By combining the DAD spectrum and the accurate mass measurement of ESI-TOF/MS, twelve components in Tanreqing injection were identified. This study contributes to clarifying the nature of Tanreqing injection, and provides an effective and reliable process for the comprehensive and systematic characterization of complex traditional Chinese medicine preparations.

NIRS-based total quality control system construction of Tanreqing injection and relative thinking / 中国中药杂志

In this paper, NIRS (near infrared spectroscopy)-based total quality control system for the Tanreqing injection was introduced briefly. By analyzing and summing up the significance and difficulties, several important problems of the practical applications which need urgent solutions are proposed. And also the applicationprospect of NIRS is fully discussed and put forward in the end.

Technological parameter optimization method for washing of coarse bear gall powder extracts based on NIR spectroscopy / 中国中药杂志

During the washing process of coarse bear gall powder extracts, it is necessary to adjust the amount of ethyl acetate according to the properties of raw materials, which aims to improving the yield and purity of the final product. In the research, using NIR spectra to reflect the comprehensive properties of coarse bear gall powder extracts, the process is optimized in a flexible way. Forty batches experiments are designed according to the weight ratio of ethyl acetate and coarse extracts of bear gall powder. The NIR spectra of the coarse extracts of bear gall powder are collected and processed using principal component analysis (PCA) method. The first 8 principal components combined with the amount of the ethyl acetate are used as the input variables, and calibration models are established to predict the yield and purity of the final product 30 batches are used as calibration set, which is used to establish the models, and other 10 batches are used as validation set, which is used for the performance appraisal of the established models. The correlation coefficients of the calibration, inner cross-validation and external validation for the purity model are 0.902, 0.896 and 0.883, respectively, and the RMSEC, RMSECV and RMSEP are 1.22%, 1.48% and 1.59%, respectively. The correlation coefficients of the calibration, inner cross-validation and external validation for the yield model are 0.921, 0.859 and 0.916, respectively, and the RMSEC, RMSECV and RMSEP are 1.39%, 1.65% and 1.53% respectively. This work demonstrated that NIR spectra combined with technology parameter could be used to predict the yield and purity of the final product. Using the established models, the most appropriate amount of the ethyl acetate can be determined according to the properties of the coarse bear gall powder extracts, and the yield and purity of the final product can be improved.

Quality control of Lonicerae japonicae flos with standard reference extract / 中国中药杂志

The high price of the reference substances is an obstacle for the HPLC analysis of Lonicerae Japonicae Flos. To solve this problem, a new method based on the standard reference extract (SRE) was proposed. In this study, the extract of Lonicerae Japonicae Flos was calibrated, and the long-term stability was investigated. Different concentration solutions of SRE were prepared for establishment of the calibration profiles, and 6 organic acids were determined. T-test was used for the comparison of the determination results via reference substances and SRE, and the results demonstrated that there is no significant difference between the two methods. The presented method can be used for the quality control of Lonicerae Japonicae Flos, and will also offer reference to resolve similar problems.

Screening and mutation of a marine Bacillus megaterium to resolute 3-chloro-1, 2-epoxypropane / 中国药学杂志

OBJECTIVE: To obtain and mutate a marine Bacillus megaterium to resolute pharmaceutical intermediate, 3-chloro-1,2-epoxypropane. METHODS: From marine sediment, a 3-chloro-1,2-epoxypropane-resolute strain was isolated and mutated by ultraviolet rays and N+ ion. RESULTS: The strain was identified as Bacillus megaterium on the basis of biophysical and biochemical test. The optimum salinity for growth was 2%, and the result of optimum salinity showed that the microbe was a typical marine microorganism. A positive mutant exhibited excellent resolution with ee value of 99% and yield of 42%. CONCLUSION: The marine bacillus megaterium shows a large potential for industrial production.

A new method for simultaneous assay of six organic acids in Lonicerae Japonicae Flos / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To establish a method of quantitative analysis of multi-components by single marker(QAMS) for quality control of Lonicerae Japonicae Flos.</p><p><b>METHODS</b>The contents of chlorogenic acid(CA) and caffeic acid(CfA) were determined, and the relative correction factors(RCF) of other organic acids were calculated, which were used for the indirect determination.</p><p><b>RESULTS</b>The RCFs for the neochlorogenic acid(NCA), 3, 4-O-dicaffeoylquinic acid(3, 4-DCA), 3, 5-O-dicaffeoylquinic acid(3, 5-DCA), and 4, 5-O-dicaffeoylquinic acid(4, 5-DCA) were 5.462, 5.689, 2.313, 2.382(to CA) and 3.941, 4.103, 1.669, 1.718(to CfA), respectively. The established method was validated in different laboratories with different high performance liquid chromatography(HPLC) instruments and different chromatographic columns; the result indicated that the reproducibility was satisfactory. There was no significant difference between the established QAMS method and external reference method (P>0.05).</p><p><b>CONCLUSION</b>The established QAMS method can be used for simultaneous determination of 6 organic acids as quality control of Lonicerae Japonicae Flos with easy available standard substances.</p>