The Study On Nanoemulsion Drug Delivery System Of Bererine

In this research, for the merits of nanotechnology, the traditional berberine was recasted by using the nanoemulsion drug delivery system as a new carrier. At first, the establishment of analytical method, screening of blank nanoemulsion prescription and the inspection of influence factor were studied. Nanoemulsion drug delivery system of berberine was prepared by pseudotertiary phase diagram. And the quality stability, security, pharmacodynamics, pharmacokinetics and release properties were systemically studied. The abstracts are as follows:1. The establishment of analytical method on nanoemulsion drug delivery system of berberine.To establish the analytical method for content determining of berberine nanoemulsion drug delivery system, Ultraviolet was used in preparation and HPLC was used in preparation and dynamic. The Ultraviolet absorption wavelength, standard curve, recovery and accuracy were studied by Ultraviolet spectrophotometer and HPLC. Results shown that berberine has the maximal absorption in 340 nm. The good linear range was 1～40μg/ml.The mean recovery is 99.7 %.RSD of the recovery is 0.39 %,the type duplication is 2.5 %,the accuracies with-day and between-day are lesss than 0.6 %.HPLC:The good linear range of the preparation was 50～1 600 ng/ml, The examination limit is 2 ng/mL, the mean recovery is 99.4% and the mean retention time is 7.499 min. RSD of the recovery is 0.79 %,the type duplication is 0.64 % and 1.44 %, the accuracies with-day and between-day are is less than 0.6 %; The good linear range of the blood plasma was 10～1 600 ng/ml.The examination limits is 0.36 ng/mL, the mean recovery is 95.6% and the mean retention time is 5.602 min. RSD of the recovery is 3.64 %, the type duplication is 0.42 % and 1.42 %,the accuracies with-day and between-day are lesss than 1%.The analysis method we established in this study have high recovery, accuracy and repeatable, it can be used in quality control and dynamics research.2. The prescription screening and the influence factors inspection of blank nanoemulsion drug delivery systemThe prescriptions of the blank nanoemulsion were screened and its influence factors were observed. Prescriptions of nanoemulsion were screened with the HLB and the nanoemulsion evaluation standard, pseudoternary phase diagrams were used to be observed the formula of nanoemulsion; the enhanced solubility was studied also. Results show that the prescriptions of screening that can form nanoemulsion are: the surfactant is: the Tween kind (80 and 60), Span kind (80 and 60), EL-40and RH-40; the cosurfactant is: the absolute ethyl alcohol, the glycerine, synth eticnbutyl alcohol, n-Butanol and 1, 3-butyleneglycol. The oil phase is: IPM and liquid paraffin. The influence factors inspection result shows that surfactant and the type and retio of the oil phase are the main and the key factors for the formula of nanoemulsion. In some distance, the higher HLB of surfactant, the bigger area of nanoemulsion is. The shorter chain of cosurfactant, the bigger area of nanoemulsion is. The HLB of the oil phase and the mixed surfactant more matches, the bigger area of nanoemulsion. The the ratio (Km) to surfactant and cosurfactant be more appropriate, the bigger area of nanoemulsion. Within limits, the amount of the medicine added increase, the area of nanoemulsion increased; the solubilization effect shows that the berberine has higher solubility in nanoemulsion than in water and micelle. The prescription of the blank nanoemulsion of being screened and the influence factors inspection results laid a good foundation for further study preparation.3. The preparation and quality evaluation on nanoemulsion drug delivery system of berberine.The nanoemulsion drug delivery system of berberine was prepared and its quality was evaluated. The suitable oil phase, surfactant and cosurfactant were choused, the nanoemulsion drug delivery system of berberine were optimized by studying the pseudoternary phase diagram. The preparation method was optimized by entrapment efficiency and drug loading as evaluation standard. Their characters were detected by electron microscope, photon correlation spectroscope, the viscosity statement, refraction and electric conductivity, the content of berberine was determined by HPLC, its'stability was also detected. Results show that the nanoemulsion drug delivery system of berberine that contains EL40-glycerine-IPM was transparent spherical bubble. Its average diameter is 43.5 nm (blank nanoemulsion is 17.9 nm); the optimal preparation method was magnetic stirring, 37 oC, and 150 r/min, disperses 12 h and the order of the third kind of medicine. The physical and chemistry characters, stability parameter, acceleration and light acceleration indicate that the quality of berberine nanoemulsion drug delivery systerm was stable, its term of validity can be conjected to be 2.5 years by long-term and the classical constant temperature experiment. The quality on nanoemulsion drug delivery system of berberine was stable. It conforms to the request of nanoemulsion drug delivery system. 4. The safety evaluation on nanoemulsion drug delivery system of berberine.To the safety evaluate of berberine nanoemulsion drug delivery system. Evaluate the safety of berberine nanoemulsion drug delivery systerm by acute toxicity, skin irritation, eye irritation (single and several), cell toxicity experiment. Results show that the maximal tolerance dose test on mice of the berberine nanoemulsion drug delivery systerm was 875mg/kg. Converts into adult's dosage (50 kg), is 21,875 times of clinical amount used. According to Bliss, counts software with SAS6.0, self-procedure, calculates its accumulation LD50 is 3 055 mg/kg. The stimulation to the party of rabbit's skin and eye are all non-toxic. The cell toxicity is 1 level, not obvious toxicity. The berberine nanoemulsion drug delivery system is non-toxic to mouse, domestic rabbit skin and eye, cell, clinical with medicine security, conforms to the request of nanoemulsion drug delivery system.5. The pharmacodynamics evaluation on nanoemulsion drug delivery system of berberine.The pharmacodynamics on nanoemulsion drug delivery system of berberine was evaluated. The pharmacodynamics on nanoemulsion drug delivery system of berberine was evaluated by antibacterial activity in vitro, preventing and treating effect on bacterial diarrhea caused by E.coli, the mice blood sugar influence by normal and high glucose tolerance, the mice serum lipids influence experiment by acute hyperlipemia. Results show that the nanoemulsion drug delivery system of berberine had the same antibacterial activity to Staphylococcus aureus, E.coli, S.agalactae, Sam onellal, its antibacterial diameter is respectively 4, 3.83, 3.83, 3.66 times as that of tablet, capsule, watery solution, and blanking nanoemulsion. Bacterial diarrhea caused by E.coli, The diarrhea times that berberine nanoemulsion decreased is respectively2,2.2,2,1.93 times(Preventing tests) and 2.7,2.13,1.88,2.22 times(Treating tests) that of watery solution, tablet, capsule, Xieliting tablets. Continuously administrated for 6d, with 50mg/kg, 1 time/d, the normal blood sugar of mice reduced.But there is no remarkable differences between each experimental group(P>0.05). high glucose tolerance model caused by ip 2.0 g/kg glucose,administere the drug as the method above, berberine nanoemulsion is respectively 4.48,3.57,3.88 times that of tablet, capsule, glibenclamide for decreasing the average of mice ralitive blood glucose(0, 30, 60, 120min). The drug and the method above were administered. The acute yperlipemia model caused by ip 0.25 ml/10 g yolk emulsion (75 %). After administrated using the method above, the reduction of the total cholesterol and the arteriosclerosis index relative value of rats caused by berberine nanoemulsion is 4.51, 2.18, 4.60:2.08, 1.06, 1.27; 2.64, 1.64, 1.44 times as that of tablet, capsule and enofibrate. The pharmacodynamics results show that the berberine nanoemulsion is better than tablet and capsule.6. The pharmacokinetics and release study on nanoemulsion drug delivery system of berberine.The pharmacokinetics and in vitro release of berberine nanoemulsion were studied. The berberine nanoemulsion was administered to the rabbit by ig with 50 mg/kg,in the time of 0,0.16,0.33,0.5,0.75,1,2,4,8,12,18,24 h, berberine was determined through HPLC in its plasma. The medicine kinematics parameter was calculated. The release behavior of berberine nanoemulsion,tablet and capsule in SGF,SIF,PBS (pH6.8)was inspected by dialysis seven normal release model carve were used to fitting the release curve. Results show that berberine nanoemulsion, tablet and capsule are all fit for the two room open model, in rabbit. The average peak time values of berberine nanoemulsion (4.205 h) enhance 2.826 h,3.088 h than tablet(1.379 h) and capsule(1.117 h).The average highest plasma concentration of its (113.699μg/L)is 1.820,1.894 times of ablet (62.466μg/L)and capsule(60.021μg/L), The relative bioavailability is339% and 332% that of tablet and capsule.The release result shows that berberine nanoemulsion fit for Hixon-crowell equation in SGF and PBS.Fit for one level release equation in SIF. The pharmacokinetics indicated that the relative bioavailability of nanoemulsion increased obviously and there is some controlled release effect. The release behavior shows that the release content of erberine nanoemulsion is highest in SIF and the release of the drug is passive diffusion.