Study On The Matrix Selection For Cultivating Bear Bile Powder Analogue In Vitro Based On Compositions Similarity

Bear bile is popularly used in traditional Chinese medicines, "Bear bile powder" is dried of the drainage of bile by the gallbladder with bile of selenarctos thibetanus curvier or Ursus arctos L., it belongs to the new medicine of Chinese medicine. With the constant and deepening development of the modernization of traditional Chinese medicine, all kinds of traditional prescriptions of bear bile and the new drug of TCM have developed rapidly, but limited to the special and scarce resources of bear bile powder, which greatly hindered the further development and the widely use of bear bile in traditional Chinese medicine. At present, looking for alternatives to bear bile powder has become a hot topic research in pharmacy, clinical medicine and other fields. This study is from "major new drugs innovation and development"(2014ZX09301306-007). In this study, given the sources and low cost of poultry bile powder, and on the basis of the methods and results of the material composition research of bear bile powder,this study used the theories and methods of multidisciplinary courses and modern instrument analysis to analyze and define the main bile acid of poultry bile powder( chicken powder, duck gall powder, geese gall powder) and other compositions of bile acid(amino acids, protein, trace elements, bilirubin, cholesterol), and then we compared all these compositions with bear bile powder is to make clear of the similarity and the main difference of poultry bile powder and bear bile powder, so as to screen out the best transformation matrix for “cultivation of bear bile powder analogue in vitro”.The main work and conclusions of this paper are as follows:(1) The content of TCDCA of poultry bile powder and bear bile powder were measured by using HPLC-ELSD The results show that: all poultry bile powder contain TCDCA, and the contents of TCDCA were: Goose bile powder(53.75%)> Silkie bile powder(49.35%)> chicken bile powder(48.5%)> duck bile powder(37.35%)> broiler bile powder(37.2%)> bear bile powder(31.0%). The contents of TCDCA of goose bile powder, silkie bile powder and chicken bile powder are greater than 38%, it accord with the requirements of the substrate in vitro simulation.(2) Taking advantage of the theories and methods of medicine, pharmacology, drug and modern instrumental analysis and other disciplines. the instrumental of amino acid analyzer, atomic spectrophotometer, UV spectrophotometer, HPLC and BCA kit methods were used for measuring the content of amino acids, trace elements, bilirubin, cholesterol and protein in each poultry bile powder and bear bile powder. The results show that:① Results of amino acid: the amino acids species of Bear bile powder and poultry are consistent, both contain 17 kinds of amino acids. Bear bile powder has only trace amounts of free amino acids and the order of the total content of hydrolytic amino acid is: bear bile powder< broiler bile powder< silkie bile powder< chicken bile powder< duck bile powder< goose bile powder.② Results of Trace Elements: Overall, the bear bile and poultry powder both contain 18 trace elements, and the content of trace elements in bear bile powder were significantly lower than those in other bile powder, the harmful metals(Pb, Cd, Be, Pr, Sr, Ba)in poultry bile powder are significantly higher than those in bear bile powder, but are lower than the definitive standard for heavy metal content prescribed by Chinese pharmacopoeia, therefore it consistent with the substrate requirements for vitro simulation.③ Bilirubin results: the content of bilirubin of broiler bile powder and bear bile powder are relatively higher than others, and no significant difference between the two. Silkie bile powder and goose bile powder are approaching, and no significant difference between the two.④ Cholesterol measurement: the order of the content of cholesterol of each sample: chicken bile powder> broiler bile powder> silkie bile powder> duck bile powder> goose bile powder> bear bile powder.⑤ The results of protein: the content of protein of goose bile powder is the highest,and there is no significant difference in broiler bile powder and bear bile powder.(3) Using Euclidean distance and generalized accard coefficient to evaluate the similarity between poutry bile powder and bear bile powder. Euclidean distance calculation results show that: the similarity between chicken bile powder and bear bile powder is the highest, 0.470; secondly,the similarity between broiler bile powder and bear bile powder is 0.454, and the similarity between silkie bile powder and bear bile powder is 0.415; ducks and geese bile powder are the lowest, the similarity are 0.389 and 0.352 respectively. The results of generalized Jaccard coefficient show that: for trace element classes, the similarity between chicken bile powder and bear bile powder is the highest; for the amino acids, bilirubin and the class, the similarity between the chicken bile powder and the bear bile powder is the highest; for the cholesterol, the similarity between goose bile powder and bear bile powder is the highest. This calculation results are consistent with the results of Euclidean distance.Based on all of the above findings: the content of TCDCA of Chicken bile powder is 45.8%, which is greater than 38%, and the similarity between chicken bile and bear bile powder is greater than other types of poultry chicken bile powder. Taken together, we choose the chicken bile powder as the best transformation matrix of " vitro cultivation of bear bile powder analogue"...