Establishment On Prediction Method For Substantial Variations In The Manufacture Of Tradition Chinese Drug (Ⅱ)

In manufacture of tradition Chinese drug(TCD), the quality of product could be influenced by many variations of manufacture. Objective evaluation on whether these variations were substantial is important in the approval and registration of TCD. Being lack of unified and objective standards, how to evaluate substantiality of the variations have become a key and difficult technical problems in the approval of new drugs. Especially, Some preparation process such as concentration, drying and shaping were normally regarded as bring about substantial variations in the quality of TCD by reason of inadequate proofs.In order to establish an objective evaluation on "substantiality of the variations" in manufacture of TCD, Radix et Rhizoma Rhei(Rhubarb) was studied as model due to its extensive application in clinic and relatively clarity of chemical components. Rhubarb Water extract by different drying techniques was investigated on physico-chemical properties and antibacterial biological activity. On this basis, the early and rapid prediction method of evaluating the variations in TCD preparation was trying to be established. Major investigation methods and results are as follow:1. The physical properties of Rhubarb water extract by different drying techniques such as appearance, pH and critical relative humidity (CRH) were investigated systematically. The results showed that appearances of different samples were notablely different, but were not obviously different in pH. The value of CRH of samples by normal press drying(69.8±5.6%) were higher than that of other samples, and there were statistical different between the samples by normal press drying and that by. spray drying (55.4±2.6%). That made a conclusion that samples by different drying techniques were stabler than those by other drying techniques. On the other hand, the results of thermo-gravimetric analysis(TG), different thermal analysis(DTA)and different scanning calorimetry(DSC)showed that samples by spray drying and microwave drying were different among all samples in the first phase of weight loss of spectrum with TG. And there were too complex component in samples by different drying techniques to sufficient information in the spectrum with DTA and DSC.2. The chemical properties of Rhubarb water extract by different drying techniques were investigated by the content of chemical components, fingerprint with ultra-performance liquid chromatogram(UPLC) and spectrum with fourier transform infrared spectrometry(FTIR). The results of determination of chemical components showed the content of total anthraquinones (3.36±0.08 %) and conjugated anthraquinones (2.34±0.10 %) in freeze-dried samples were highest , compared with samples by normal press drying a statistically significant(p<0.05). But the content of free anthraquinones and tannins in all samples were little different, compared with each other no statistically significant.(p>0.05). According to information reflected by UPLC fingerprints and FTIR spectrum, samples by different drying techniques could be divided into three sorts by through of the drying conditions.3. Antibacterial biological activity of samples by different drying techniques was investigated preliminarily with biothermodynamics. First at all, the experimental parameters of biothermodynamics methods, such as culture medium and bacteria were optimized. the results of experiment with optimal parameter show growth of E. coli were inhibited after adding Rhubarb water extract by different drying techniques ,with. compared with blank group. Growth rate parameter in second logarithmic growing phase(k2) could stand for overall patterns of information in biothermodynamics, and dose-effect relationship was significant with the parameter k2 (Y =-0.00036 X+ 0.01546,r =0.9524,p =0.0109). The above all explained that antibacterial bioactivity or biopotency of Rhubarb water extract by different drying technique could be evaluated by through of the information which k2 reflected in biothermodynamics. Afterwards, difference in antibacterial biological activity of different samples was investigated. According to the results of k2 and similarity of biothermodynamics curves (S), k2 of samples by freeze drying were higher than that of samples by other drying methods. S was no statistically significant, compared with each othe(rp>0.05). And there was some difference in k2 of different samples. These come to the conclusion that antibacterial activity could be used as an important method of investigating difference in biological effects of samples by different drying conditions.4. The relativities and difference in the results by different dective methods were synthetically compared with. By means of it, so was an program for evaluation on substantial of the variations in the manufacture of TCD, with based on common requirement of physico-chemical properties., analysis of multiple key components, and analysis of biothermodynamics which constitute a predicting sequel, which could provide new research ideas and technical support for the development and approval of new drug.