The Study On Modern Processing Method And Uplc-dad-tof-ms Fingerprint Of Wen-hou-po

Objective: Cortex Magnoliae Officinalis is a widely used TCM, which is documented in Chinese Pharmacopoeia (2005 edition). Wen-Hou-Po, as one of the famous-region drug of cortex Magnoliae Officinalis, is a main source of cortex Magnoliae Officinalis and primarily produced in Zhejiang province. At present, the processing method of Wen-Hou-Po used in factories is not very suitable, and the quality control means for both herbs and processed drugs of Wen-Hou-Po need to be consummated. To solve these problems, we take studies on modern processing method and fingerprint of Wen-Hou-Po.Methods: The moistening method was firstly optimized by One Way ANOVA. And then, orthogonal design L9(34) was used to screen out the best processing conditions of four different processing methods (stir-frying with ginger juice, boiling with ginger juice, baking with ginger juice and microwave heating with ginger juice). After this, only one and the optimum processing method was defined through comparing the four optimized processing methods with the total content of magnolol and honokior as index. SPME-GC-MS was used to analyze the volatile components of Wen-Hou-Po herbs and its processed drugs (processed according to the optimum processing method). The stuctures, types and the relative content were given, which showed the variation trends of the volatile components after processing. Wen-Hou-Po herbs and processed drugs (processed according to the optimum processing method) from two species in five areas in Zhejiang Province were analyzed to establish two fingerprints for Wen-Hou-Po herbs and processed drugs (processed according to the optimum processing method). Data were evaluated statistically using similarity analysis, cluster analysis and discriminant analysis in order to establish fingerprints for quality control of Wen-Hou-Po herbs and processed drugs (processed according to the optimum processing method).Results: Through comparison, the optimum moistening time was 60 min, and the optimum volume of ginger juice to dosage of herbs ratio was 60 L/100 Kg. The results of four optimized processing methods were as follows, microwave heating with ginger juice (31.95 mg·g^-1) > baking with ginger juice (31.79 mg·g^-1) > boiling with ginger juice (30.55 mg·g^-1) > stir-frying with ginger juice (29.72 mg·g^-1) > commercial drugs (20.07 mg·g^-1), and the difference are not remarkable. The optimized method, boiling with ginger juice, was chosen as the optimum processing method. The result of SPME-GC-MS analysis showed that, the volatile components of Wen-Hou-Po herbs and its processed drugs were in the same types, but in different content. The absolute and relative contents of isolimonene rised after processed, while other components reduced to varying extent. The composition of the volatile oil changed greatly. Through the fingerprinting analysis of Wen-Hou-Po herbs and its processed drugs (processed according to the optimum processing method), which were from two species in five areas in Zhejiang Province, the similarity standard for Wen-Hou-Po herbs was 0.95, while the similarity standard for Wen-Hou-Po processed drugs (processed according to the optimum processing method) was 0.93. The result of hierarchical cluster analysis showed that the samples from two species in five areas could be divided into two distinct groups (the same as the groups of the samples divided by their species) based on their compositional fingerprints. And then, two rapid and convenient discriminant functions were established to discriminate the species of unknown Wen-Hou-Po, and the cross validation result was 100%. By comparing the UV and MS data with those of the authentic standards and literature, some common peaks in the fingerprints were identified.Conclusion: In this study, the optimum processing method for Wen-Hou-Po was established; the effect of the optimum processing method to volatile components was summarized; two UPLC-DAD-TOF-MS fingerprints were established for quality control of Wen-Hou-Po herbs and its processed drugs (processed according to the optimum processing method); based on the results of cluster analysis and discriminant analysis, two species-identified methods for Wen-Hou-Po herbs and its processed drugs were established, which could provide some reference for further studies.