| Realgar is a common mineral medicine, realgar and the Chinese medicines which contain realgar play an important role in the traditional Chinese medicine. As it is used in medicine it is insoluble, resulting in oral overdosage and low bioavailability, which limits the development of realgar and the Chinese medicine containing realgar in the international market.For the reasons above, in this paper, a high activity strain of bacteria named Acidithiobacillus ferrooxidans (At.f), which was isolated and identified by the professor Li Hongyu’s lab of Lanzhou university, was used to bioleach realgar, and the bioleachate of this strain was then made into an injection. This solved the problems of oral overdosage and low bioavailability. Analysis of realgar bioleachate by the method of capillary electrophoresis was used to determine its composition. Pharmacodynamics of the realgar injection were also studied preliminarily. The results are as follows:1. Four arsenic species were analyzed rapidly by capillary electrophoresis with direct UV and indirect UV detection. The arsenic compounds identified were iAsIII, iAsV, MMAV and DMAV. The optimum conditions are as follows: (1) The background electrolyte ( BGE ) of capillary electrophoresis analysis with direct UV and the separation conditions: 10mM borate, 30 kV, 25℃, pH 9.50, 195 nm, 0.5 psi, 5 s; (2) Capillary Electrophoresis with Indirect UV analysis and separation of BGE conditions: borate 10mM with CTAB 0.5mM,-20kV, 25℃, pH 9.50, 195 nm, 0.5 psi, 5 s. We used two analytical methods for realgar bioleachate and found that the species in realgar bioleachatewere iAsIII, iAsV. 2. The stability of realgar bioleachate was studied, and it was found that heat and light had a great impact on the realgar bioleachate. Thus, we chose antioxidant and chelating agents as the accessories of realgar injection in order to increase the stability of realgar injection. Orthogonal design was used to optimize the realgar injection. The quality of self-made realgar injection was tested, it was showed that quality of self-made realgar injection was in line with the requirements of injection.3. The protein, tannin, oxalate, resins, potassium ion and residue of realgar injection were tested, showed that impurity of realgar injection was in line with the general inspection requirements. After heat and light stability tests of the realgar injection, color of the samples did not change significantly, the pH value and content is relatively stable over 10 days, which indicate that high temperature and light had no effect on the stability for the physical and chemical agents in short period. Inspection of the safety of agents through the hemolysis test, acute toxicity test, abnormal toxicity test and pyrogen test showed that realgar injection achieved met the desired criteria.4. The biological activity of realgar injection was studied, the results of inhibition of tumor cells in vitro was tested and showed by an inhibiting rate of approximately 30% for human leukemia cells K562, hepatocellular carcinoma cell HepG2, gastric cancer cells MGC803, and Lewis lung cancer cells. The realgar injection have a better inhibition in vivo, with an inhibition rate for mice sarcoma S180 up to 33.85% using a 0.03mg / g realgar injection. |
