Studies On The Preparation Of The Lyophilization Injection Of Hydrochloric Liensinine

Objective: To investigate the formula of lyophilization injection of hydrochloric linesinine and technique of preparation, to control its quality comprehensively, to make the quality standard of liensinine raw material and lyophilization for injection; to study the bio-security of lyophilization injection of hydrochloric liensinine including irritability, hemolysis and vas stimulation; to develop a RP-HPLC method for the preliminary study of fingerprint on phenolated alkaloids in plumula nelumbinis and quantitatively analyze the content of liensinine, isoliensinine and neferine, which are the three main phenolated alkaloids in plumula nelumbinis.Methods: The lyophilization injection of hydrochloric liensinine with mannitol, glucose, lactose as skeleton excipients was prepared and its content was detected by RP-HPLC method(CH3CN: H2O: Triethylamine (adjust pH to 3.3 with H3PO4)=25: 75: 0.1 as mobile phase and UV detection at 282nm, Flow rate was 0.8ml min-1 and 20 l was injected every time); the freeze-drying curve was obtained when the lyophilization injection formula was produced in GMP workshop; the physical and chemical properties including content, relative matter and stability of the injection were evaluated. Guinea pigs were irritated then attacked, level of anaphylaxis was observed comparing with negative and positive control groups; the lyophilization was also investigated to show the effect to 2% red cell sedimentation solution and stimulation to the vein of rabbit ear, the local reaction of injection-spots were observed by naked eyes and vein tissues were sliced up to show the pathological change in microscope. RP-HPLC was applied to the preliminary study of fingerprint.Results: The optimum formula included liensinine 5mg and mannitol 160mg, prepared for the solution of 2ml per ampoule to be lyophilized. Stability study showed the lyophilization injection of hydrochloric liensinine had a better stability in the shadow and the liensinine raw material was stable under high temperature, but had great hygroscopicityand can be affected seriously by strong light. The two concentration solutions of liensinine showed negative irritability to guinea pig and had no coagulative and hemolysis effect to red cell. The vein of rabbit ear had an intact endodermis and showed no dilation, denaturalization putrescence. In the fingerprint, 16 common peaks of phenolated alkaloids were confirmed. The quantitative analysis showed the three main phenolated alkaloids' content, with liensinine (40.36 0.49)%, isoliensinine (44.46 0.91)% and neferine (2.62 0.11)%.Conclusions: The formula of lyophilization for injection of hydrochloric liensinine is rational, the preparation method can be applied in lab and also provide gist for workshop production. The quality standard is reliable, which can control quality of lyophilization injection of hydrochloric liensinine. Bio-security tests showed Liensinine has no irritability, no effect of coagulation and hemolysis and no significant stimulative influence to vein. The RP-HPLC method is simple and accurate with a good reproducibility and specificity and the preparatory study of fingerprint can be used scientifically as a quality control method for extract of the phenolated alkaloid in plumula nelumbinis.