绞股蓝纳米乳的制备及其抗衰老作用的研究
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摘要
为了更好的开发中药绞股蓝,本实验制备了绞股蓝纳米乳并考察了其理化性质、稳定性和安全性。主要内容如下:
     1.绞股蓝纳米乳的制备及其理化性质
     为了制备出符合药用要求的绞股蓝纳米乳。利用转相法制备纳米乳,通过滴定法绘制伪三元相图,研究其处方的组成并考察了温度、Km和离子浓度对纳米乳形成的影响;并研究其对绞股蓝的增溶作用,最终筛选出最佳配方,利用最佳配方制备出绞股蓝纳米乳,考察了其外观、电导率、折光率、粘度、形态及粒径等理化性质。结果:配方RH40/正丁醇/乙酸乙酯/蒸馏水纳米乳区最大,所制备的绞股蓝纳米乳为O/W型纳米乳,其外观为淡黄色、澄清透明、可见蓝色乳光、流动性好的液体;其电导率、折光率、粘度分别为105.6±1.23μs/cm、1.427、25mPa.S;投射电镜下观察其形态为圆球形,分布均匀,粒径介于40-60nm。绞股蓝纳米乳经透射电镜观察、理化性质考察、粒度分布检测,其各项指标符合纳米乳的特性。
     2.绞股蓝纳米乳的稳定性
     为了考察绞股蓝纳米乳的稳定性。通过影响因素试验、恒温加速试验和留样观察试验考察了绞股蓝纳米乳的稳定性。影响因素试验表明:绞股蓝纳米乳在各种条件下放置10天,外观仍为淡黄色、透明均一、流动性良好的液体,未见分层和油滴,未发生明显改变;光照及40℃条件下,含量下降,粒径无明显变化,光照试验对pH影响最大,可能是由于光照能促进磷脂的氧化分解所致。因此,本品对热、光都敏感,应于适宜温度下避光贮存,不得冷冻。恒温加速试验和留样观察试验显示:绞股蓝纳米乳各项指标基本无变化,稳定性良好。
     3.绞股蓝纳米乳的安全性
     为了考察绞股蓝纳米乳的安全性。通过小鼠急性毒性试验、家兔眼部刺激试验和细胞毒性试验考察了绞股蓝纳米乳的安全性。急性毒性试验显示绞股蓝纳米乳对2种性别小鼠的急性毒性半致死浓度为LD50 > 10 g/kg,符合药用要求。眼部刺激试验按评分标准判断其刺激程度积分为2,属轻度刺激。细胞毒性试验显示绞股蓝纳米乳对细胞毒性为1级,无明显毒性。结果表明绞股蓝纳米乳安全,无明显的毒性和刺激性。
     4.绞股蓝纳米乳的抗衰老作用
     为了研究绞股蓝纳米乳的抗衰老作用,通过测定健康老龄(18个月)SD大鼠血清中超氧化物歧化酶(SOD)活性及丙二醛(MDA)含量发现,绞股蓝纳米乳可极显著提高SOD活性和极显著降低MDA含量(p<0.01),表明绞股蓝纳米乳具有抗衰老作用。
     综上所述:本实验所制备的纳米乳符合纳米乳的各种特性,安全、稳定,符合药用要求。
To develop GP for better,GP nanoemulsion was prepared and its physical and chemical properties,stability and security was also tested.The chief content was followed:
     1. The preparation of GP nanoemusion and Its physical and chemical properties
     To prepare GP namoemulsion which according to the medicinal requirments.The namoemulsion was prepared by phase inversion emulsification.the pseudo-tertiary diagram was protractedby titration,the influence of temperature,Km and ion density on the prescription of nanoemulsion was stydied,the solubilization of GP was also studied,the optimun formula,the physical and chemical properties of its appearance conductivity refractive index biscosity and diameter distribution was measured.Results:The nanoemulsion region of RH40/Butanol/Ethyl acetate/distilled water was the maximum;the type of GP nanoemulsion was O/W,the apperance of GP namoemulsion was clarify light yellow liquid with good flowability;the conductivity refractive index viscosity of GP namoemulsion was 105.6±1.23μs/cm,1.427, 25mPa.S,the morphological character of GP namoemulsion was spheroidal,in uniform distribution,the range of its diameter was 40~60nm.conclusion:All the indexes of GP namoemulsion according to the peculiarities of namoemulsion after observed under transmission electron microscope,surveyed of its phsical and chemical properties and detected of its range of diameter.
     2. The stability of GP namoemulsion
     To study stability of GP namoemulsion .the stability of GP namoemulsion was studied through influence factors test,constant temperature accelerated test.Results: The infuence factors test indicate: GP namoemulsion was, after deposited 10 days,still clarify light yellow liquid with good flowability,had no layer and oil,below light exposure,the diameter of GP namoemulsion had no distinct change,Its content decreace but not distinct,The pH altered distinct by light exposure,the cause maybe phospholipids oxygenolysis .So,It should storage lucifuge in appropriate temperature.the constant temperature acceleratec test and the stability test indicate:the index of GP namoemulsion had no change.conclusion:the stability of GP namoemulsion was good,but It should storage lucifuge in appropriate temperature.
     3. The security of GP namoemulsion
     To study the security of GP namoemulsion.The security of GP namoemulsion was studied through acute toxiciy and cytotoxic test.Result:The acute toxicity indicate:the half lethal concentration LD50>10g/kg,according to the medicinal requirements.The integrations of stimuli was mild.The cytotoxic test indicate:the cytotoxic of GP namoemulsion was 1 degree,the cytotoxic was not distinct.
     4. The anti-aging effect of GP namoemulsion
     To study the anti-aging effect of GP namoemulsion.The activity of SOD and the content of MDA was measured in aging SD wistar.Result:compared with control group,the GP namoemulsion 10g/L group and 5g/L group can remarkably decrease the content of MDA and remarkably enhance the activity of SOD ,Its anti-aging effect approach ginsensoide group.The results indicate GP namoemulsion had anti-aging effect,the mechanisms maybe relate with clear away free-radical and anti oxidation.
     Above all,the GP namoemulsion was according to the peculiarity of namoemulsion,safe and stable,in accordance with the requirements of medicine and had ramarkable anti-aging effect.
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