The Authenticity Identification Of The Traditional Chinese Medicine Bezoar And Its Substitutes And The Quality Control Method Of The Bezoar Medicinal Material And The Patent Medicine

Bovis Calculus is a dry gallstone obtained from the gallbladder or bile duct of bovine(Bos taurus domesticus Gemlin).It is a pathological product of cattle and is very precious.Because Calculus Bovis is expensive and rare,there is a high demand in the market.Chinese scholars have developed Calculus Bovis substitutes-in vitro cultured Calculus Bovis and cultivated Calculus Bovis,artificial Calculus Bovis and so on.Chinese Pharmacopoeia(2015 edition)contains Calculus Bovis,artificial Calculus Bovis and in vitro cultured Calculus Bovis.Because the prices of Calculus Bovis and its substitutes is different(natural Calculus Bovis 300000 ~ 400000 yuan / kg,cultivated in vitro,4 0000~ 50,000 yuan / kg,artificial Calculus Bovis,several hundred yuan / kg),In recent years,there has been a situation in the market in which the adulteration of Calculus Bovis is adulterated to seek illegal benefits,the use of cheap Calculus Bovis substitutes to replace or mix with Calculus Bovis,and the insufficient feeding of Calculus Bovis and its substitutes in the production of proprietary Chinese medicine,these problems seriously affect the quality of traditional Chinese medicine and traditional Chinese medicine,and threaten the safety and health of people to use medicine.Therefore,it is necessary to identify and distinguish Calculus Bovis and its substitutes(cultivated Calculus Bovis,in vitro cultured Calculus Bovis,and artificial Calculus Bovis),and to control and evaluate the quality of traditional Chinese medicine and patent medicine in order to solve the problems in the Calculus Bovis market and to ensure the safety and effectiveness of peopleundefineds drug use.This paper mainly includes the following four parts:Part ? Progress in research on chemical constituents and quality control methods of Calculus Bovis and its substitutes.The chemical composition and quality control method of Calculus Bovis and its substitute are reviewed in this paper.Through the study of literature,the chemical components of Calculus Bovis,cultivated Calculus Bovis and in vitro cultured Calculus Bovis were similar,but the proportion of different types of bilirubin was different.At present,the specific components of Calculus Bovis have not been found to distinguish and,in vitro cultured Calculus Bovis.At present,the specific components of Calculus Bovis have not been found to distinguish Calculus Bovis from,in vitro cultured Calculus Bovis and cultivated Calculus Bovis.Part ? To establish a method for the identification of bezoar mixed substitutes(mainly in vitro cultured Calculus Bovis).Progenesis QI statistical analysis software combined with multivariate data analysis method was used to distinguish Calculus Bovis and its two substitutes,and then partial least square(OPLS-DA)was used to identify the specific characteristic components of Calculus Bovis and in vitro cultured Calculus Bovis.The characteristic components found by the QQQ-MS instrument with higher sensitivity were verified to avoid false negative results.Finally,various animal bile powder and bezoar powder close to the Calculus Bovis component are added for final verification,and the specificity of the characteristic components is confirmed.This method can accurately distinguish Calculus Bovis from its substitutes,and can also detect whether it is mixed with cheaper substitutes in precious Chinese patent medicine containing Calculus Bovis.Part ? On the basis of 20 batches of Calculus Bovis samples,the characteristic total ion chromatogram(TIC)of Calculus Bovis was established.According to the scanning mode of precursor ion and its daughter ion,the transmission voltage and collision voltage of the parent ions of five chemical components of Calculus Bovis were optimized.Two stable ion pairs of each parent ion were obtained,and ten sets of ion pairs were established.The characteristic chromatogram of the chemical constituents of Calculus Bovis were established by using the(MRM)mode of mass spectrometry and the data collection of ten groups of ion pairs.Part ? Multi-index quality control and analysis of natural Calculus Bovis,in vitro cultured Calculus Bovis medicinal material and its patent medicine..In this study,a simple UPLC-QQQ-MS method was established to determine the contents of 13 kinds of bile acids in Calculus Bovis,in vitro cultured Calculus Bovis,Xihuang pills containing Calculus Bovis and Xihuang pills containing in vitro cultured Calculus Bovis.Verified by methodology,Based on the analysis of bile acids,the quality of Calculus Bovis,in vitro cultured Calculus Bovis and Xihuang pills containing Calculus Bovis(or in vitro cultured Calculus Bovis)were evaluated,and their fingerprints of bile acids were established.The results showed that the total content of bile acids(total 13 bile acids)in vitro cultured Calculus Bovis was significantly higher than that of Calculus Bovis.With the free bile acid-cholic acid(especially cholic acid)as the index,the content and proportion of cholic acid in vitro cultured Calculus Bovis and its Xihuang pills were significantly higher than those of Calculus Bovis and its Xihuang pills.According to the content of combined bile acid(taurine-binding bile acids and glycine-binding bile acids),most of Calculus Bovis and its Xihuang pill were slightly higher than those of in vitro cultured Calculus Bovis and its Xihuang pill.