| Calculus bovis,cattle gallstones, is a precious Chinese herbal medicines. The main chemical composition of bile acids, deoxycholic acid, chenodeoxycholic acid, bilirubin, etc., with heat, phlegm,resuscitation, and detoxification effect. Calculus bovis cultivated by glucuronidase is a kinds of calculus bovis,as the raw material is fresh bile formation under the principle of natural calculus bovis, by enzyme preparation.The method of fingerprint for calculus bovis cultivated by glucuronidase and natural calculus bovis has established. Through systematic research in the bile acid, bilirubin, cholesterol, trace elements and other components on consistency with the evaluation of new drugs for the calculus bovis cultivated by glucuronidase development and product quality control standards to provide experimental evidence and technical information. Main contents and results of research are as follows:1.The studies of qualitative and quantitative in the DMSO extracts of calculus bovis cultivated by glucuronidase and natural calculus bovis.The HPLC fingerprint of natural calculus bovis with a comparative study of calculus bovis cultivated by glucuronidase was established. The fingerprint of calculus bovis cultivated by glucuronidase has high similarity, suggesting that the production of calculus bovis cultivated by glucuronidase more stable. However, a total of 10 natural calculus bovis peaks, can be detected 5 peaks in calculus bovis cultivated by glucuronidase, most of them less than the peak signal strength of natural calculus bovis. The main component of the calculus bovis cultivated by glucuronidase lower than natural calculus bovis.But the calculus bovis cultivated by glucuronidase also has a component higher than natural calculus bovis. Calculus bovis cultivated by glucuronidase and natural calculus bovis that DMSO extracts of the type and content were different.The HPLC method for determination of bilirubin was established. C18 column, methanol-chloroform-1% acetic acid (65:32:3) mobile phase, the detection wavelength 450 nm, flow rate 1.0 mL·min-1. The content of calculus bovis cultivated by glucuronidase and natural calculus bovis were 9.35μg·mg-1,45.32μg·mg-1, bilirubin are quite different.2.The studies of qualitative and quantitative on bile acids of calculus bovis cultivated by glucuronidase and natural calculus bovis.The HPLC-ELSD fingerprint was established with calculus bovis cultivated by glucuronidase and natural calculus bovis..The software of Traditional Chinese Medicine fingerprint similarity evaluation system (version 2004A) were used for data proeessing.The fingerprint similarity of calculus bovis cultivated by glucuronidase was 0.980,0.991,0.979,0.990,0.984,0.992,0.983,0.989,0.991 and 0.983,indicating that the higher similarity between different batches of calculus bovis cultivated by glucuronidase.It has a stable production. Lower similarity in common pattern natural calculus bovis and calculus bovis cultivated by glucuronidase were detected. Constituents of natural calculus bovis can be detected eight acid peaks,but five peaks of calculus bovis cultivated by glucuronidase can be detected, which the four and five peaks no less than the peak signal strength of natural calculus bovis. The three,sewen and eight peaks were not detected in calculus bovis cultivated by glucuronidase, but the nine,ten and eleven peaks of natural calculus bovis not detected, show that the composition of acids were different.The HPLC-ELSD method of six bile acids was established. C18 column, ELSD detector, methanol and 0.1% acetic acid mobile phase gradient elution. Bile acid, chenodeoxycholic acid and deoxycholic acid were detected in natural calculus bovis; but bile acid,chenodeoxycholic acid and hyodeoxycholic acid detected bezoar were detected in calculus bovis cultivated by glucuronidase, that is equal to the composition of two acid types, one different, and differences in content, the detection of three batches of 3 kinds of two calculus bovis acid content of ingredients are basically the same class.3.The comparison of trace elements in calculus bovis cultivated by glucuronidase and natural calculus bovisThe 12 elements of calculus bovis were determinated by flame atomic absorption spectrometry and graphite furnace atomic absorption spectrometry. The small difference of Cu, Zn, Fe, Mn, Ca, Mg, K, Na 8 elements and Pb, Cr, Ni, Cd heavy metals were compared with calculus bovis cultivated by glucuronidase and natural calculus bovis. The comparative study of trace elements for the further development and utilization to provide a scientific basis for calculus bovis cultivated by glucuronidase.4.The quantitative comparison of cholesterol in calculus bovis cultivated by glucuronidase and natural calculus bovisThe HPLC method for determination of cholesterol was established. C18 column, ELSD detector, acetonitrile and isopropanol (1:1) as mobile phase.The content of the three batches of natural calculus bovis was 6.53μg·mg-1,5.69μg·mg-1,5.12μg·mg-1, but the content of three batches of calculus bovis cultivated by glucuronidase was 2.78μg·mg-1,2.34μg·mg-1,2.13μg·mg-1.The high content of cholesterol, suggesting that may be related to the natural mechanism of the body into a stone on.
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