The Research On Preparation Craft Of Total Alkali Of Harmel Chitosan Nanoparticle As Injection

Objective:To develop the brain target responsive total alkali of harmel nanoparticle on the base of screening the extraction and purification program of total alkali of harmel and its pharmacologic action. Methods:1) The content of harmine and harmaline were selected as index, the extraction and purification program of total alkali of harmel was optimized by the uniform design.2) The huperzine A was selected as control drug, the effects of total alkali of harmel on ability of remembering and the content of SOD in two kind AD rats, induced by AlCl3 and by D-galactose, was observed. And the relationship of dose-effect was studied.3) The particle diameter of nanoparticle and encapsulation efficiency were selected as index, the program of cross-linking methods of harmel total alkali chitosan nanoparticle was optimized by the orthogonal design.4) The brain target effect of total alkali of harmel nanoparticle was evaluated according to the relative absorption rate (re), target effect (te) and the maximal concentration ratio (Ce). Results:1) The optimized extracting process is as following:adding 4 times 30% alcohol to the seed, extracting time lasting 7 hours. And the preliminary purification process is as following: adding 4 times 0.5% HCl to rude extracts, ultrasonic time lasting 10 minutes, adding ammonia water to adjust pH 9, filtering and drying. The last purification process is as following:adding 3 times 95% alcohol to the coarse powder and extracting 15 minutes, filtering and drying. The total content of harmine and harmaline in the purificated extracts was 87.3%.2) The difference of remembering ability and the content of SOD between experimental groups and model group (P<0.01), but there are no obviously difference between experimental groups and model group (P>0.05). It shows that total alkali of harmel has therapeutical effects on the aphronesia mice animal model induced by AlCl3and D- galactose. The best dosage of mice is 33mg/kg.3) The preparation program of nanoparticle is as following:the 20mg chitosan was dissolved in 20ml glacial acetic acid and the NaOH was added to adjust pH 4(Ⅰ). The sodium tripolyphosphate (TPP) 6mg and Tween-80 15mg were dissolved in 5ml water (Ⅱ). Then (Ⅱ) was slowly added to (Ⅰ) and the mixture was stirred 0.5h (700r/min). The mean diameter of nanoparticle is 96nm, encapsulation efficiency is 43.75%, loading dosage is 29.17%, zeta electric potential is +26.33mv.4) The re of total alkali of harmel nanoparticle in brain is 2.46. The maxium value of te is 2.88 and the minum value of te is 1.34. The Ce of total alkali of harmel nanoparticle in brain is 2.48. Conclusions:The optimized process of the extraction and purification is available, reasonable, and reproducible. The effects of total alkali of harmel on the aphronesia mice were obviously observed. And the results of verification test shows that the preparation program of total alkali of harmel chitosan nanoparticle is available, reasonable. The total alkali of harmel chitosan nanoparticle has obvious brain target effect.