Studies On Improvement Of Quality Standard Of Pheretima Aspergillum And Standard Decoction

Objective:To supplement the identification method of Pheretima aspergillum,establish the content determination,fingerprint detection and analysis method of Pheretima aspergillum,determine the preparation process of Pheretima aspergillum standard decoction,and study the quality standard of Pheretima aspergillum standard decoction,so as to provide some reference for the research of Pheretima aspergillum related quality standard.Methods:1.The DNA barcoding technology was used to identify the basic source of the collected samples.Combined with the methods of characteristic,microscopic identification and TLC identification in the 2015 edition of Chinese pharmacopoeia,the samples used in the research were identified in the 2015 edition of Chinese pharmacopoeia.2.The single-factor investigation of liquid chromatography conditions and test solution was performed,and the contents of hypoxanthine and inosine in 30 batches of Guangdilong medicinal materials were determined by high performance liquid chromatography(HPLC).Simultaneously establish HPLC fingerprints of 30 batches of Guangdilong.3 To extract rate and hypoxanthine and inosine content transfer rate of multiple indicators of water,decoction time,decoction times factors such as the single factor investigation,and select the best factor level,orthogonal experiment was carried out,and the specific detail parameters,through the orthogonal experiment data analysis,screening to determine preparation technology parameters determined by process validation test process.4.Collect 13 batches of Pheretima aspergillum samples to prepare Pheretima aspergillum standard decoction with the determined preparation process,determine p H value,total solids and content,and calculate the transfer rate of index components;At the same time,the fingerprint of Pheretima aspergillum standard decoction was studied.Results:1 DNA barcoding technology can effectively identify the basic source of the collected samples and the results are accurate.2.The results of the assay methodology showed that the linear ranges of hypoxanthine and inosine were 1.58 m L ~ 31.6 m L /m L(r=0.999 9)and 5.52 m L ~ 110.4 m L(r=0.999 8),respectively.RSD of precision,stability and repeatability test was all less than 2.0%(n=6).The average recovery was 103.0 %(RSD=1.7 %,n=6)and 101.2 %(RSD=1.2 %,n=6),respectively.HPLC fingerprints of 30 batches of Guangdilong samples were successfully established.By comparing with the reference spectra,12 common peaks were matched and 6 components were identified.Among them,the HPLC fingerprints of 28 batches were similar to the HPLC control fingerprints.Degree> 0.900.3.The preparation process was as follows: 100.0g of Pheretima aspergillum was taken,soaked in 8 times of water for 30 min,refluxed for 20 min,filtered while it was hot,added 6 times of water to the residue,refluxed for 20 min,filtered while it was hot,combined with 3 times of filtrate,reduced pressure and concentrated to 500 m L,then the Pheretima aspergillum standard decoction was obtained.4.In this study,13 batches of Pheretima aspergillum standard decoction were prepared,of which the transfer rate of total subxanthine and inosine ranged from 78.7 to 101.8 %,the p H value ranged from 5.6 to 6.0,and the total solids ranged from 0.22 to 0.48 g.The fingerprints of 13 batches of Pheretima aspergillum standard decoction were established.Conclusion:The DNA barcoding technology is accurate and reliable,and the DNA barcoding technology can be used for the identification of the base source of radix lumbricolae.The HPLC fingerprint and the method for the determination of hypoxanthine and inosine were established.The optimized preparation process of Pheretima aspergillum standard decoction is stable,reliable and repeatable,and the research on the quality standard of Pheretima aspergillum standard decoction is carried out to provide reference and data support for the research on the forms of traditional Chinese medicine granule and the establishment of the standardization of traditional Chinese medicine,which is conducive to the realization of unified supervision and management.