当归、川芎挥发油静脉给药微乳研究及体内评价
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摘要
挥发油是中药中常见的有效部位，不仅含量较高，药理作用也非常明确，在抗心脑血管疾病、抗肿瘤、抗病毒等方面发挥了重要作用。但是因为中药挥发油在水中的溶解度太小，静脉给药制剂的开发一直是药剂学的难题。本文中选用当归、川芎组方后经水蒸气蒸馏法提取的挥发油作为模型药，研究能够静脉给药用O/W型微乳，并对其理化性质、体内分布和药物动力学进行了研究。
微乳(Microemulsion，ME)作为静脉给药载体研究已经成为药剂学领域一个新的热点。与乳剂、脂质体和胶团溶液等相比，微乳具有热力学稳定，可热压灭菌，也可微膜滤过；粘度低，注射时不引起疼痛；粒径低于血红细胞的直径；药物能缓慢释放，延长药效并具有靶向作用等特点，是一种有广泛前景的药物载体。但是由于微乳的制备过程中采用了大量的乳化剂和助乳化剂，对人体有潜在的毒副作用，成为限制其应用的最大障碍。因此在选用高效低毒乳化剂和助乳化剂的基础上，通过伪三元相图法研究微乳的形成条件，通过优化工艺，寻找用最少的乳化剂和助乳化剂制备微乳的新方法，并能够避免注入体内被血液稀释而导致的微乳相转变和微乳粒径变大等现象的发生。
藁本内酯是当归和川芎挥发油中共同含有的指标性成分，含量较高，药效显著，但是在室温条件下很容易被降解，极其不稳定，因此在进行研究前，首先采用硅胶柱层析方法经过二次洗脱制备其对照品，并用IR、UV、NMR、MS光谱对其结构进行鉴定，确认为Z-藁本内酯，纯度大于98％，并以此建立了高效液相色谱方法用于藁本内酯的含量测定。
在进行处方研究之前，先对水溶液中导致藁本内酯不稳定的各方面因素进行了探讨，明确光照是导致藁本内酯降解的最主要因素，需要严格避光保存，另外，贮存温度、载体系统的组成、pH值和稳定剂的加入等均能在不同程度上影响藁本内酯的降解速度。同时，对不同载体系统中藁本内酯的降解趋势进行拟合，发现其符合Weibull概率分布。实验发现，能够使藁本内酯充分稳定的载体系统中含有20％的1，2-丙二醇和0.30％的稳定剂S，溶剂系统的pH值调至5.8，并向溶液和容器中通入惰性气体，整个操作过程和贮存时需要严格避光。经典恒温法加速实验和Arrhenius方程推算藁本内酯在上述载体系统中室温条件下放置的贮存期(T_(90))为1.77年，室温留样观察为1.75年，二者基本一致，为含有藁本内酯的液体制剂的开发提供参考。
本课题中研究的模型药物是当归、川芎经水蒸气蒸馏法提取得到的挥发油，其中碳链长度在10～12之间的内酯类成分占60％以上，本身就可以做为微乳的油相，避免了另外加入其它油相引起的潜在的毒性，同时预实验也证明，加入大豆油等其他的油相成分必须同时增加乳化剂和助乳化剂的使用量，这必将进一步增加制剂的刺激性和毒性。
本课题在文献报道能够静脉注射的四种乳化剂(磷脂、泊洛沙姆188、Cremophor EL35和Solutol HS15)中通过绘制伪三元相图进行乳化剂的选择，结果发现Solutol HS15所制备微乳的状态最佳，而且其能够提高微乳的稳定性，延长药物在血液循环中的时间，有利于本制剂的开发。助乳化剂确定为无水乙醇，且乳化剂与助乳化剂的质量比锁定为1：1，可以防止液晶态和凝胶相的形成，而且所得微乳的粘度较小，适合于注射给药。
综合微乳处方研究的伪三元相图中得到的信息，确定微乳处方中乳化剂/助乳化剂与挥发油的比例保持在7：3时，所制备的微乳区域可延伸到水相的顶点，即可用水大量稀释而不会发生微乳相转变。
本课题在研究过程中，考察了改变微乳中的含水量时，相应样本的粘度、电导率、折光率的变化，并对其相转变过程进行了初步探讨，确定出乳化剂和助乳化剂含量不变情况下，增溶最大量挥发油的组成点，并确定微乳经过大量稀释后粒径不会变大。外界温度的变化对微乳区域的形状有一定的影响，但是在伪三元相图中存在着相同一部分区域，可以认为该区域内受外界温度变化的影响不明显，操作过程中可以忽略其变化以简化过程。而以生理盐水替代双蒸水所制备得到的伪三元相图并没有区别，证明增加离子性添加物并不会影响微乳的形成。透射电镜显示，所制备的微乳为球形的粒子，平均粒径在110nm左右，经过生理盐水稀释后，粒径明显减小。
采用HPLC方法测定微乳中藁本内酯的含量，并以藁本内酯含量、粒径、电导率、折光率、粘度为指标对微乳的理化稳定性进行考察，加速及长期留样6个月，当归、川芎挥发油微乳基本稳定。
Bliss法计算当归、川芎挥发油微乳小鼠尾静脉注射单次给药的半数致死量(LD_(50))为321.04mg/kg，95％置信区间为239.44～430.45mg/kg。
建立了2倍体积的乙腈蛋白沉淀法处理血浆和生物组织样品，对当归、川芎挥发油微乳在家兔体内的药物动力学和小鼠体内组织分布进行了考察，方法学考察符合体内药物分析的要求。通过家兔耳缘静脉注射当归、川芎挥发油微乳和1，2-丙二醇溶液，发现微乳给药后，在家兔体内分布符合二室模型，t_(1/2α)为1.37min，t_(1/2β)为10.48min，AUC为28051.56ng min/ml；而当归、川芎挥发油的1，2-丙二醇溶液注射给药后很快消除，4min后检测不到药物。藁本内酯在小鼠体内的组织分布考察表明，当归、川芎挥发油的1，2-丙二醇溶液给药后藁本内酯很快从血浆中消除，血浆内测到的藁本内酯的最高浓度仅为微乳得到的1/6左右，其药时曲线下的面积(AUC)仅为75.09μg．min/g，而微乳的AUC为946.71μg．min/g，而藁本内酯主要浓集于肾脏。说明微乳给药后藁本内酯在血浆内的循环时间明显延长，AUC显著增大，而且体内各个组织器官中分布较为均匀，没有发生器官浓集现象。藁本内酯在小鼠血浆中分布符合二室模型，与家兔体内结果基本一致。
当归、川芎挥发油静脉注射给药后藁本内酯在家兔和小鼠血浆内的药物动力学研究表明，虽然以微乳为载体能较大程度上提高其在血浆内的滞留时间，但其半衰期仍较短，难以维持有效治疗浓度，因此提示，在临床上可以将微乳用生理盐水稀释到适当的浓度后采用静脉滴注的形式给药，有利于发挥药效并减少刺激性。
Volatile oil is one of the common parts in Traditional Chinese Medicines with great contents and noticeable pharmacological effects, which is used extensively in some diseases such as cardio-cerebrovascular, tumor and virus aspects. However, because of poor solubility for most volatile oil, studies about its praeparatum through veins have great difficulties. In this study, the volatile oil extracted by steaming distillation from Danggui and Chuanxiong was chosen as model drug to develop an O/W microemulsion vehicle for intravenous administration, what is more, the physical and chemical properties, pharmacokinetics in rabbits and distribution in mice were investigated, too.
Microemulsion has become one of the hot subjects of applied study for intravenous administration in pharmaceutics domains. Comparing to emulsion, liposome and micellar solution, drug delivery system of microemulsion offers considerable potential predominance as follows: it is a thermodynamically stable, isotropic transparent or semitransparent system, so it can pass through micropore filter, stand autoclave sterilization; the viscosity of microemulsion is rather low, which would lessen much pain during injection; the particle diameters of microemulsion are usually less than the that of red blood cells; the drugs entrapped in microemulsion will be released slowly, at the same time, it maybe has some targeting effects. However, the large amount of surfactant and cosurfactant in composition of microemulsion is the main barrier which blocks its more application in intravenous administration. Therefore, after selecting high effective and low toxic surfactant and cosurfactant, the forming condition of microemulsion can be optimized by pseudo-ternary phase diagrams to obtain a formula with less surfactant and cosurfactant and more volatile oil. At the same time, the microemulsion prepared should prevent phase transition when it is diluted by blood after injection.
Z-ligustilide is the primary ingredient both in volatile oil of Danggui and Chuanxiong. But its extreme instability at room temperature restricts its application in pharmacy. In this study, before pharmaceutical experiments, the standard substance of Z-ligustilide was separated by gel silica column chromatography through elution twice, and its purity was more than 98% by HPLC. The structure was identified by IR, NMR, MS and UV methods to be Z-ligustilide.
As mentioned before, Z-ligustilide is rather instable, and illumination is the major factor which induced its degradation. Other factors, which could affect its stability in aqueous solutions including pH value, composition of solvents, kinds and amounts of stabilizer, etc al, were detailedly investigated by accelerating experiments. The results indicated that the most suitable vehicle for Z-ligustilide included 20% 1, 2-propanediol and 0.30% stabilizer S, and its pH value was adjusted to 5.8. The degradation trend of Z-ligustilide was fitted according with Weibull probability distribution, and its shelf-life(T_(90)) was 1.77 years through Arrhenius equation by classical thermal methods, which was accordant to the facts.
As the model drug in this study was volatile oil from Traditional Chinese Medicines, in which the lactone with carbon chain between 10～12 was more than 60%, it may serve as the oil phase of microemulsion per se, which could avoid the toxicity of other oils. Meanwhile, experimental results presented that, the addition of other oils such as soybean oil needs more surfactant and cosurfactant to form microemulsion, thus could increase the potential danger of preparations.
Four surfactants including phospholipids, Poloxamer 188, Cremophor EL 35 and Solutol HS 15, which could be adopted by intravenous injection as reported were considered by pseudo-ternary phase diagrams, and results showed that Solutol HS 15 was more suitable to prepare microemulsion, furthermore, the microemulsion containing Solutol HS 15 could improve its stability and prolong existing time in blood circulation, which was more favorable for this preparation. The cosurfactant was ethanol and the mass weight ratio of surfactant to cosurfactant was fixed at 1:1, which could prevent appearance of liquid crystal and gels. The viscosity of microemulsion prepared is low enough for injection.
According to all the information obtained form pseudo-ternary diagrams, the weight ratio of surfactant/cosurfactant to volatile oil was maintained as 7:3, thus, the O/W microemulsion domain could extended to the culminated point of water phase in diagrams, which suggested that the microemulsion would not occur phase inversion even was diluted by large amount of mediums.
During study, the viscosity, electric conductivity, refractivity and particle diameters of samples containing different amount of water were reviewed respectively, and according to these results, the phase transition process was approached initially. When the amount of surfactant and cosurfactant was fixed, the composition with maximum volatile oil was determined. Moreover, the particle diameters would not enlarge when it was diluted. The surrounding temperatures would affect the shape and size of O/W microemulsion in diagrams to a certain degree, however, there existed some common domains under different temperatures, which could be predicated to be less influenced by temperature, and this could simplify the preparation process. The O/W microemulsion domain had no change when the distilled was substituted by normal saline, which proved ionic addictives would not affect the formation of microemulsions. The photo under transmission electron microscope displayed the microemulsion drops as spherical particles with average diameter of 110nm, and dilution by normal saline or water would reduce its diameters.
The content of Z-ligustilide in microemulsion was determined by HPLC. The stability of microemulsion was investigated by viscosity, electric conductivity, refractivity, particle diameters and content of Z-ligustilide and results of accelerating testing(40℃) and room temperature showed it was basically stable during 6 months.
Through single intravenous injection to mice, the median lethal dosage(LD50) of microemulsion was 321.04mg/kg, and the 95% confidence interval was 239.44～430.45mg/kg treated by Bliss procedure.
There were few reports about the in vivo study about Z-ligustilide in volatile oil of Danggui and Chuanxiong. A protein precipitation method using 2 times of acetonitrile was developed to treat the samples of plasma and biological tissues, and the pharmacokinetics of Z-ligustilide in rabbits and tissue distribution in mice were investigated by intravenous injection. The distribution of Z-ligustilide in microemulsion was fitted a two-compartment model in rabbits, and t_(1/2α) and t_(1/2β) was 1.37min and 10.48min, respectively, and the area under curves(AUC) was 28051.56 ng min/ml. However, when the volatile was administrated as 1, 2-propanediol solution, no drug could be detected after 4min. The results of tissue distribution in mice showed that AUC of microemulsion in plasma was 12 times more than that of solution, and the distribution of Z-ligustilide in every tissue was uniform. However, the concentration of Z-ligustilide in plasma was only sixth to that of microemulsion soon after the volatile oil solution was injected; other drugs were found mostly enriched in kidney. The distribution of Z-ligustilide in mice plasma was fitted a two-compartment model, which was accordant to that in rabbits.
As the basis of pharmacokinetics of Z-ligustilide both in rabbits and mice after the volatile oil microemulsion was administrated by intravenous injection, it was found that, although the existence of Z-ligustilide in plasma had been greatly improved, it would be eliminated from plasma quickly. This fact meant that it was difficult for Z-ligustilide to remain an effective concentration. Thus, the volatile oil microemulsion was recommended to be administrated by intravenous infusion after it was diluted to certain concentration, and this method might profit the pharmacodynamic action and reduce its stimulation.
微乳(Microemulsion，ME)作为静脉给药载体研究已经成为药剂学领域一个新的热点。与乳剂、脂质体和胶团溶液等相比，微乳具有热力学稳定，可热压灭菌，也可微膜滤过；粘度低，注射时不引起疼痛；粒径低于血红细胞的直径；药物能缓慢释放，延长药效并具有靶向作用等特点，是一种有广泛前景的药物载体。但是由于微乳的制备过程中采用了大量的乳化剂和助乳化剂，对人体有潜在的毒副作用，成为限制其应用的最大障碍。因此在选用高效低毒乳化剂和助乳化剂的基础上，通过伪三元相图法研究微乳的形成条件，通过优化工艺，寻找用最少的乳化剂和助乳化剂制备微乳的新方法，并能够避免注入体内被血液稀释而导致的微乳相转变和微乳粒径变大等现象的发生。
藁本内酯是当归和川芎挥发油中共同含有的指标性成分，含量较高，药效显著，但是在室温条件下很容易被降解，极其不稳定，因此在进行研究前，首先采用硅胶柱层析方法经过二次洗脱制备其对照品，并用IR、UV、NMR、MS光谱对其结构进行鉴定，确认为Z-藁本内酯，纯度大于98％，并以此建立了高效液相色谱方法用于藁本内酯的含量测定。
在进行处方研究之前，先对水溶液中导致藁本内酯不稳定的各方面因素进行了探讨，明确光照是导致藁本内酯降解的最主要因素，需要严格避光保存，另外，贮存温度、载体系统的组成、pH值和稳定剂的加入等均能在不同程度上影响藁本内酯的降解速度。同时，对不同载体系统中藁本内酯的降解趋势进行拟合，发现其符合Weibull概率分布。实验发现，能够使藁本内酯充分稳定的载体系统中含有20％的1，2-丙二醇和0.30％的稳定剂S，溶剂系统的pH值调至5.8，并向溶液和容器中通入惰性气体，整个操作过程和贮存时需要严格避光。经典恒温法加速实验和Arrhenius方程推算藁本内酯在上述载体系统中室温条件下放置的贮存期(T_(90))为1.77年，室温留样观察为1.75年，二者基本一致，为含有藁本内酯的液体制剂的开发提供参考。
本课题中研究的模型药物是当归、川芎经水蒸气蒸馏法提取得到的挥发油，其中碳链长度在10～12之间的内酯类成分占60％以上，本身就可以做为微乳的油相，避免了另外加入其它油相引起的潜在的毒性，同时预实验也证明，加入大豆油等其他的油相成分必须同时增加乳化剂和助乳化剂的使用量，这必将进一步增加制剂的刺激性和毒性。
本课题在文献报道能够静脉注射的四种乳化剂(磷脂、泊洛沙姆188、Cremophor EL35和Solutol HS15)中通过绘制伪三元相图进行乳化剂的选择，结果发现Solutol HS15所制备微乳的状态最佳，而且其能够提高微乳的稳定性，延长药物在血液循环中的时间，有利于本制剂的开发。助乳化剂确定为无水乙醇，且乳化剂与助乳化剂的质量比锁定为1：1，可以防止液晶态和凝胶相的形成，而且所得微乳的粘度较小，适合于注射给药。
综合微乳处方研究的伪三元相图中得到的信息，确定微乳处方中乳化剂/助乳化剂与挥发油的比例保持在7：3时，所制备的微乳区域可延伸到水相的顶点，即可用水大量稀释而不会发生微乳相转变。
本课题在研究过程中，考察了改变微乳中的含水量时，相应样本的粘度、电导率、折光率的变化，并对其相转变过程进行了初步探讨，确定出乳化剂和助乳化剂含量不变情况下，增溶最大量挥发油的组成点，并确定微乳经过大量稀释后粒径不会变大。外界温度的变化对微乳区域的形状有一定的影响，但是在伪三元相图中存在着相同一部分区域，可以认为该区域内受外界温度变化的影响不明显，操作过程中可以忽略其变化以简化过程。而以生理盐水替代双蒸水所制备得到的伪三元相图并没有区别，证明增加离子性添加物并不会影响微乳的形成。透射电镜显示，所制备的微乳为球形的粒子，平均粒径在110nm左右，经过生理盐水稀释后，粒径明显减小。
采用HPLC方法测定微乳中藁本内酯的含量，并以藁本内酯含量、粒径、电导率、折光率、粘度为指标对微乳的理化稳定性进行考察，加速及长期留样6个月，当归、川芎挥发油微乳基本稳定。
Bliss法计算当归、川芎挥发油微乳小鼠尾静脉注射单次给药的半数致死量(LD_(50))为321.04mg/kg，95％置信区间为239.44～430.45mg/kg。
建立了2倍体积的乙腈蛋白沉淀法处理血浆和生物组织样品，对当归、川芎挥发油微乳在家兔体内的药物动力学和小鼠体内组织分布进行了考察，方法学考察符合体内药物分析的要求。通过家兔耳缘静脉注射当归、川芎挥发油微乳和1，2-丙二醇溶液，发现微乳给药后，在家兔体内分布符合二室模型，t_(1/2α)为1.37min，t_(1/2β)为10.48min，AUC为28051.56ng min/ml；而当归、川芎挥发油的1，2-丙二醇溶液注射给药后很快消除，4min后检测不到药物。藁本内酯在小鼠体内的组织分布考察表明，当归、川芎挥发油的1，2-丙二醇溶液给药后藁本内酯很快从血浆中消除，血浆内测到的藁本内酯的最高浓度仅为微乳得到的1/6左右，其药时曲线下的面积(AUC)仅为75.09μg．min/g，而微乳的AUC为946.71μg．min/g，而藁本内酯主要浓集于肾脏。说明微乳给药后藁本内酯在血浆内的循环时间明显延长，AUC显著增大，而且体内各个组织器官中分布较为均匀，没有发生器官浓集现象。藁本内酯在小鼠血浆中分布符合二室模型，与家兔体内结果基本一致。
当归、川芎挥发油静脉注射给药后藁本内酯在家兔和小鼠血浆内的药物动力学研究表明，虽然以微乳为载体能较大程度上提高其在血浆内的滞留时间，但其半衰期仍较短，难以维持有效治疗浓度，因此提示，在临床上可以将微乳用生理盐水稀释到适当的浓度后采用静脉滴注的形式给药，有利于发挥药效并减少刺激性。
Volatile oil is one of the common parts in Traditional Chinese Medicines with great contents and noticeable pharmacological effects, which is used extensively in some diseases such as cardio-cerebrovascular, tumor and virus aspects. However, because of poor solubility for most volatile oil, studies about its praeparatum through veins have great difficulties. In this study, the volatile oil extracted by steaming distillation from Danggui and Chuanxiong was chosen as model drug to develop an O/W microemulsion vehicle for intravenous administration, what is more, the physical and chemical properties, pharmacokinetics in rabbits and distribution in mice were investigated, too.
Microemulsion has become one of the hot subjects of applied study for intravenous administration in pharmaceutics domains. Comparing to emulsion, liposome and micellar solution, drug delivery system of microemulsion offers considerable potential predominance as follows: it is a thermodynamically stable, isotropic transparent or semitransparent system, so it can pass through micropore filter, stand autoclave sterilization; the viscosity of microemulsion is rather low, which would lessen much pain during injection; the particle diameters of microemulsion are usually less than the that of red blood cells; the drugs entrapped in microemulsion will be released slowly, at the same time, it maybe has some targeting effects. However, the large amount of surfactant and cosurfactant in composition of microemulsion is the main barrier which blocks its more application in intravenous administration. Therefore, after selecting high effective and low toxic surfactant and cosurfactant, the forming condition of microemulsion can be optimized by pseudo-ternary phase diagrams to obtain a formula with less surfactant and cosurfactant and more volatile oil. At the same time, the microemulsion prepared should prevent phase transition when it is diluted by blood after injection.
Z-ligustilide is the primary ingredient both in volatile oil of Danggui and Chuanxiong. But its extreme instability at room temperature restricts its application in pharmacy. In this study, before pharmaceutical experiments, the standard substance of Z-ligustilide was separated by gel silica column chromatography through elution twice, and its purity was more than 98% by HPLC. The structure was identified by IR, NMR, MS and UV methods to be Z-ligustilide.
As mentioned before, Z-ligustilide is rather instable, and illumination is the major factor which induced its degradation. Other factors, which could affect its stability in aqueous solutions including pH value, composition of solvents, kinds and amounts of stabilizer, etc al, were detailedly investigated by accelerating experiments. The results indicated that the most suitable vehicle for Z-ligustilide included 20% 1, 2-propanediol and 0.30% stabilizer S, and its pH value was adjusted to 5.8. The degradation trend of Z-ligustilide was fitted according with Weibull probability distribution, and its shelf-life(T_(90)) was 1.77 years through Arrhenius equation by classical thermal methods, which was accordant to the facts.
As the model drug in this study was volatile oil from Traditional Chinese Medicines, in which the lactone with carbon chain between 10～12 was more than 60%, it may serve as the oil phase of microemulsion per se, which could avoid the toxicity of other oils. Meanwhile, experimental results presented that, the addition of other oils such as soybean oil needs more surfactant and cosurfactant to form microemulsion, thus could increase the potential danger of preparations.
Four surfactants including phospholipids, Poloxamer 188, Cremophor EL 35 and Solutol HS 15, which could be adopted by intravenous injection as reported were considered by pseudo-ternary phase diagrams, and results showed that Solutol HS 15 was more suitable to prepare microemulsion, furthermore, the microemulsion containing Solutol HS 15 could improve its stability and prolong existing time in blood circulation, which was more favorable for this preparation. The cosurfactant was ethanol and the mass weight ratio of surfactant to cosurfactant was fixed at 1:1, which could prevent appearance of liquid crystal and gels. The viscosity of microemulsion prepared is low enough for injection.
According to all the information obtained form pseudo-ternary diagrams, the weight ratio of surfactant/cosurfactant to volatile oil was maintained as 7:3, thus, the O/W microemulsion domain could extended to the culminated point of water phase in diagrams, which suggested that the microemulsion would not occur phase inversion even was diluted by large amount of mediums.
During study, the viscosity, electric conductivity, refractivity and particle diameters of samples containing different amount of water were reviewed respectively, and according to these results, the phase transition process was approached initially. When the amount of surfactant and cosurfactant was fixed, the composition with maximum volatile oil was determined. Moreover, the particle diameters would not enlarge when it was diluted. The surrounding temperatures would affect the shape and size of O/W microemulsion in diagrams to a certain degree, however, there existed some common domains under different temperatures, which could be predicated to be less influenced by temperature, and this could simplify the preparation process. The O/W microemulsion domain had no change when the distilled was substituted by normal saline, which proved ionic addictives would not affect the formation of microemulsions. The photo under transmission electron microscope displayed the microemulsion drops as spherical particles with average diameter of 110nm, and dilution by normal saline or water would reduce its diameters.
The content of Z-ligustilide in microemulsion was determined by HPLC. The stability of microemulsion was investigated by viscosity, electric conductivity, refractivity, particle diameters and content of Z-ligustilide and results of accelerating testing(40℃) and room temperature showed it was basically stable during 6 months.
Through single intravenous injection to mice, the median lethal dosage(LD50) of microemulsion was 321.04mg/kg, and the 95% confidence interval was 239.44～430.45mg/kg treated by Bliss procedure.
There were few reports about the in vivo study about Z-ligustilide in volatile oil of Danggui and Chuanxiong. A protein precipitation method using 2 times of acetonitrile was developed to treat the samples of plasma and biological tissues, and the pharmacokinetics of Z-ligustilide in rabbits and tissue distribution in mice were investigated by intravenous injection. The distribution of Z-ligustilide in microemulsion was fitted a two-compartment model in rabbits, and t_(1/2α) and t_(1/2β) was 1.37min and 10.48min, respectively, and the area under curves(AUC) was 28051.56 ng min/ml. However, when the volatile was administrated as 1, 2-propanediol solution, no drug could be detected after 4min. The results of tissue distribution in mice showed that AUC of microemulsion in plasma was 12 times more than that of solution, and the distribution of Z-ligustilide in every tissue was uniform. However, the concentration of Z-ligustilide in plasma was only sixth to that of microemulsion soon after the volatile oil solution was injected; other drugs were found mostly enriched in kidney. The distribution of Z-ligustilide in mice plasma was fitted a two-compartment model, which was accordant to that in rabbits.
As the basis of pharmacokinetics of Z-ligustilide both in rabbits and mice after the volatile oil microemulsion was administrated by intravenous injection, it was found that, although the existence of Z-ligustilide in plasma had been greatly improved, it would be eliminated from plasma quickly. This fact meant that it was difficult for Z-ligustilide to remain an effective concentration. Thus, the volatile oil microemulsion was recommended to be administrated by intravenous infusion after it was diluted to certain concentration, and this method might profit the pharmacodynamic action and reduce its stimulation.
引文
1．王靖，张铁军，王文燕．羟丙基-β-环糊精对降香挥发油包合作用的研究[J]．中草药，2006，37(5)：700-703
2．李文志，冯汝就．鸦胆子油乳注射液联合TACE治疗原发性肝癌的临床疗效观察[J]．中药材，2006，29(6)：632-633
3．王云杰，黄立军，张志培，王小平．康莱特注射液治疗原发性肝癌的临床研究[J]．现代肿瘤医学，2006，14(1)：36-37
4．禹玉洪，李雪春，吴涛，屠鹏飞．注射用柴胡挥发油脂质体制备工艺研究[J]．中国中药杂志，2004，29(6)：521-525
5．罗琥捷，李临生．鱼腥草挥发油纳米脂质体的制备[J]．陕西中医，2005，26(12)：1370-1371
6．姚世霞，倪京满，钱松，任凤娇．当归挥发油脂质体制备工艺研究[J]．中成药，2007，29(1)：54-57
7．韩静，唐星，巴德纯．降香挥发油固体脂质纳米粒的制备工艺研究[J]．沈阳药科大学学报，2004， 21 (4): 257-260, 296
8. Malcolmson C, Lawrence M J. A comparison of incorporation of model steroids into non-ionic micellar and microemulsion systems [J]. Journal of Pharmacy and Pharmacology. 1993, 45 (1): 141-143
9. Hoar T P, Schulman J. Transparent water in oil dispersions: oleopathic hydromicelle [J]. Nature (Lond.), 1943, 152:102-103
10. Danielsson I, Lindman B. The definition of a microemulsion [J]. Colloids and Surfaces, 1981, 3: 391-392
11．张正全，陆彬．微乳给药系统研究概况[J]．中国医药工业杂志，2001，32(3)：139-142
12. Winsor P A. Solvent properties of amphiphilic compounds [M]. London, Butterworth's Scientific publication, 1954
13．郭荣，李干佐．阴离子型微乳液的电导行为及其溶液结构[J]．化学学报，1987，45(1)：55-58
14．李泉，李维红，翁诗甫，吴瑾光，徐光宪．水／AOT／正庚烷微乳体系中水结构的FT-IR研究[J]．应用化学，1998，15(1)：1-4
15. Lindman B, Moseley M E, Stilbs P. Fourier-transform NMR self-diffusion and microemulsion structure [J]. Journal of Colloid and Interface Science, 1981, 83 (2): 569-582
16. Jahn W. Srtrey R. Microstructure of microemulsions by freeze fracture electron Microscopy [J]. Journal of Physical Chemistry, 1988, 92:2294-2301
17. Cebula D J, Myers D Y, Ottewill R H. Studies on microemulsions Part 1: Scattering studies on water-in-oil microemulsions [J]. Colloid and Polymer Science, 1982, 260 (1): 96-107
18. Hou M J, Kim M, Shah D O. A light scattering study on the droplet size and interdroplet interaction in microemulsions of AOT-oil-water system [J]. Journal of Colloid Interface Science, 1988, 123 (2): 398-412
19. Attwood D, Ktistic G. A light scattering study on oil-in-water microemulsions [J]. International Journal of Pharmaceutics, 1989, 52 (2): 165-171
20. Shinoda K, Friberg S. Microemulsions: colloidal aspects [J]. Advances in colloid and interfaces science, 1975, 4:281-300
21. Bidner M S, Larson R G, Scriven L E. On the theory of micellization with reference to etroleum recovery [J]. Rev.Latinoam.Ing.Quim.Apl., 1976, 6 (1): 1-32
22．哈润华，侯斯健，栗付平．微乳液结构和丙烯酰胺反相微乳液聚合[J]．高分子通报，1995，1：10-19
23. Prince L M. Microemulsion, theory and practice [M]. Chapter 5, New York: Academic press, 1997.
24. Shinoda K, Araki M, Sadaghiani A, Khan A, Lindman B. Lecithin based microemulsions: phase behavior and microstructure [J]. The Journal of Physical Chemistry, 1991, 95:989-993
25. Aboofazeli R, Lawrence M J. Investigation into the formation and characterization of phosphor-lipids microemulsions. Ⅰ. Pseudo-ternary phase diagrams of systems containing water-lecithin-alcoholisopropylmyristrate [J]. International Journal of Pharmaceutics, 1993, 93: 161-175
26. Milan J S, Katrin S. Microemulsion in technical processes [J]. American Chemical Society, 1995, 95: 849-864
27．吴顺芹，李三鸣，赵国斌．微乳及其在药剂学中的应用[J]．沈阳药科大学学报，2003，20(5)：381-385
28．鲁莹，刘英，蒋雪涛．新型药物载体：微乳[J]．国外医药——合成药 生化药 制剂分册，1999，20(4)：253-258
29．涂秋榕，陈洁，朱家璧．自乳化药物传递系统[J]．国外医药——合成药 生化药 制剂分册，2002，23(3)：170—173
30．崔晶，翟光喜，邹满．微乳给药系统的研究与应用[J]．食品与药品，2005，7(1A)：23-26
31. M. Jayne Lawrence, Gareth D. Rees. Microemulsion based media as novel drug delivery systems [J]. Advanced Drug Delivery Reviews. 2000, 45:89-121
32．王晓黎，蒋雪涛．微乳在药剂学上的应用[J]．解放军药学学报，2000，16(2)：88-91
33．王倩，丁红梅，周海源．微乳相图及工艺研究[J]．西北药学杂志，1997，12(2)：72-73
34．陈华兵，翁婷，杨祥良．微乳在现代药剂学中的研究进展[J]．中国医药工业杂志，2004，35(8)：502-506
35．杨惊宇，严冬，罗英杰，杨大坚，陈士林．新型药物剂型——微乳[J]．中国医学工程，2005，13(4)：378-381
36．陈耀祖，陈能煜，马学毅，李海泉．当归化学成份分析研究——毛细管气相色谱-质谱法鉴定当归根挥发油成份[J]．高等学校化学学报，1984，5(1)：124-128
37. Zhao K J, Dong T, Tu P F, Tsim KWK. Molecular genetic and chemical assessment of Radix angelica (Danggui) in China [J]. Journal of Agricultural and Food Chemistry, 2003, 51: 2576-2583
38．闫升，乔国芳，刘志峰，刘珂，王嘉陵．当归油对大鼠离体子宫平滑肌收缩功能的影响[J]．中草 药，2000，31(8)：604-606
39．刘卫，刘玉玲，吕泰省，刘传绩，俞惠琴．芎归滴丸对海马神经元钠电流的保护作用[J]．中国药学杂志，2002，37(1)：824-826
40．孙所，俞惠琴．芎归滴丸对大鼠脑血流量的影响[J]．山东医药工业，1999，18(4)：44-45
41．郭书英，刘卫，俞惠琴，赵晓民，高允生，苏定冯．芎归滴丸对选择性去窦弓神经大鼠动脉压力感受反射敏感性的影响[J]．中国药房，2003，14(6)：329-331
42. Hou Y Z, Zhao G R, Yang J, Yuan Y J, Zhu G G, Hiltunen R. Protective effect of Ligusticum Chuanxiong and Angelica Sinensis on endothelial cell damage induced by hydrogen peroxide [J]. Life Science, 2004, 75:1775-1786
1．胡长鹰，丁霄霖．当归中藁本内酯的提取、分离与结构鉴定[J]．无锡轻工大学学报，2003，22(5)：69-71
2. Kaouadji M, Pachtere F D, Paouget C, Chulia A J. Three additional phthalide derivatives, an epoxymonomer and two dimers, from Ligusticum wallichii rhizomes [J]. Journal of Natural Products, 1986, 49 (5): 872-877
3．路新华，梁鸿，赵玉英．当归中藁本内酯类化合物的分离与结构鉴定[J]．中国中药杂志，2003，28(5)：423-425
4．宋治中，崔育新，朱启秀，贾忠建．二维H-NMR研究双藁本内酯结构[J]．波谱学杂志，1991，8(2)：209-214
5．宋金春，罗顺德，蔡鸿生，徐锋．反相高效液相色谱法测定生化汤中阿魏酸及其血药浓度[J]．中国医院药学杂志，2001，21(4)：213-215
6．王映芬，何燕萍，蔡阳．复方偏头痛冲剂主要成份当归、川芎中阿魏酸的含量测定[J]．北京医科大学学报，1994，26(3)：218-218
7．贾晓斌，施亚芳，黄一平，上官方．当归和川芎中阿魏酸高效液相色谱测定方法的改进[J]．中成药，1998，20(6)：37-38
8．常明向，刘小平．当归、川芎及配伍后煎煮液中阿魏酸含量的变化[J]．中药饮片，1992，2：24-25
9．梁明金，贺浪冲．藁本内酯和丁烯酜内酯对bFGF诱导的大鼠平滑肌细胞异常增殖的抑制作用[J]．药学学报，2006，41(2)：161-165
10．石力夫，郑晓梅，蔡溱，吴柏生．藁本内酯分解前后川芎挥发油对兔球结膜微循环影响的比较 [J]．中国药理学与毒理学杂志，1995，9(2)：157-158
11．汪程远，杜俊荣，钱忠明．藁本内酯的研究进展[J]．中国药学杂志，2006，41(12)：889-891
12．金灯萍，彭国平，陆晓峰．川芎中藁本内酯对照品的制备[J]．中草药，2006，37(1)：64-65
13．马建龙，李汉蕴，戴荣继，邓玉林．当归化学成分藁本内酯的提取纯化[J]．药品评价，2005，2(3)：214-215
14．苗爱东，梁乾德，刘勇，周喆，王升启．SPE-HPLC制备色谱法从当归挥发油中分离纯化Z-藁本内酯[J]．中药材，2005，28(5)：778-779
15. Li Huabin, Chen Feng. Preparative isolation and purification of chuanxiongzine from the medicinal plant Ligusticum Chuanxiong by high-speed counter-current chromatography [J]. Journal of chromatography A, 2004, 1047:249-253
16．张达磊，李桂生，任召言，曲桂武．气相色谱法测定川芎挥发油中Z-藁本内酯及川芎内酯A的含量[J]．药物分析杂志，2006，26(7)：895-897
17．汪程远，张浩，钱忠明．HPLC测定不同川芎药材中藁本内酯[J]．中草药，2006，37(6)：447-449
18．苗爱东，梁乾德，王升启．SPE-HPLC测定四物合剂中藁本内酯的含量[J]．中药材，2005，27(6)：737-738
19．赵志鸿，刘晨江．薄层荧光扫描法测定参茸近视康复片中藁本内酯的含量[J]．中国实验方剂学杂志，1997，3(2)：4-6
20．王爱武，邱福军，吕文海．双波长薄层扫描法测定当归散中藁本内酯的含量[J]．中国医院药学杂志，2003，23(1)：31-32
21．周长新，邹建凯，陈耀祖，赵昱．气相色谱-质谱法测定当归挥发油中藁本内酯的含量[J]．药物分析杂志，2002，22(4)：290-292
22. Lin Longze, He Xianguo, Lian Lizhi, King Wayne, Elliott Jerry. Liquid chromatographic-electrospray mass spectrometric study of the phthalides of Angelica Sinensis and chemical changes of Z-ligustilide [J]. Journal of Chromatography A, 1998, 810:71-79
23．梁忠明，曹铁城．试述中药提取工艺和新进展[J]．时珍国药研究，1993，4(4)：36-38
24．黄保民．中药提取分离工艺中高新技术应用的进展[J]．中医研究，1998，11(5)：56-58
25．李菁，葛发欢，黄晓芬，李莹，辉国钧．超临界CO_2萃取当归挥发油的研究[J]．中药材，1996，19(4)：187-189
26．胡志方，郭慧玲．川芎、当归挥发油成分混合提取工艺的探讨[J]．江西中医学院学报，2004，16(3)：49-50
27．李桂生，马成俊，刘志锋，刘珂．超临界CO_2萃取法与水蒸气蒸馏法提取当归挥发油的比较[J]．中草药，2001，32(7)：579-583
28．李桂生，刘岩，刘珂．水蒸汽蒸馏法对当归挥发油提取过程中成分异构化的影响[J]．中成药，2001，23(11)：784-786
1．陈耀祖，陈能煜，马学毅，李海泉．当归化学成份分析研究——毛细管气相色谱．质谱法鉴定当归根挥发油成份[J]．高等学校化学学报，1984，5(1)：124-128
2．黄远征，溥发鼎．川芎根茎挥发油化学成分的研究[J]．药学学报，1988，23(6)：426-429
3．石立夫，邓延昭，吴柏生．川芎干燥根茎挥发油化学成分及其稳定性的研究[J]．药物分析杂志，1995，15(3)：26-30
4．李桂生，马成俊，李香玉，刘珂．藁本内酯的稳定性研究及异构化产物的GC-MS分析[J]．中草药，2000，31(6)：405-407
5．周长新，李新华．藁本内酯的稳定性与溶剂化效应的关系[J]．药学学报，2001，36(10)：793-795
6. Bohrmann H, Stahl E, Mitsuhashi H. Constituents of umbelliferous plants (ⅩⅢ). Chromatographic studies on the constituents of cnidium officinale [J]. Chemical and Pharmaceutical Bulletin, 1967, 15:1606-1608
7．李慧，王一涛．藁本内酯稳定性的影响因素及稳定化措施[J]．江西中医学院学报，2003，15(1)：56-57
8．王曦，张曼红，孙杰．硝酸甘油注射液的制备[J]．山东医药工业，1999，18(5)：9-10
9．姜洪志，陆忠贤．苯巴比妥钠注射液贮存期预测[J]．中国药房，1991，2(4)：23-23
10. Naoya Okusa. Prediction of stability of drugs. Ⅲ. Application of Weibull probability paper to prediction of stability [J]. Chemical and Pharmaceutical Bulletin, 1975, 23 (4): 794-802
11．张伟德，叶兴凯．金属离子在液相氧化中的催化作用[J]．分子催化，1993，7(3)：203-211
12. Kobayashi M, Mitsuhashi H. Studies on constituents of Umbelliferae plants. ⅩⅦ. Structures of three new ligustilide derivatives from Ligusticum wallichii [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (12): 4789-4792
13. Kaouadji M, Pouget C, Pouget C. Three additional phthalide derivatives, an epoxymonomer and two dimers, from Ligusticum wallichii rhizomes [J]. Journal of Natural Products. 1986, 49 (5): 872-877
14. Tsuchida T, Kobayashi M, Kaneko K, et al. Studies on the constituents of Umbelliferae plants. ⅩⅥ. Isolation and structures of three new ligustilide derivatives from Angelica acutiloba [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (11): 4460-4464
15. Kobayashi M, Fujita M. Studies on constituent of Umbelliferae plants. ⅩⅤ. Constituents of Cnidium officinale: Occurrence of pregnenolone, coniferylferulate and hydroxyphthalides [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (4): 1427-1433
16. Lin Longze, He Xianguo, Lian Lizhi, King Wayne, Elliott Herry. Liquid chromatographic electrospray mass spectrometric study of the Angelica sinensis and chemical changes of Z-ligustilide [J]. Journal of Chromatography A, 1998, 810:71-79
17．杜金山．光线与药物的稳定性[J]．天津药学，1995，7(2)：33-35
18．Moore D，郑集声．药物光降解作用原理和实验研究[J]．国外医药药学分册，1998，3：174-177
19．乔传武，东星月，吕秀荣．防止或延缓药物水解保证药物质量的疗效[J]．中国误诊学杂志，2003，3(3)：454-454
20．王仿成．制剂因素与药物质量[J]．安庆医学，2000，21(4)：195—195
21．Mchael JA．药剂产品中的稳定剂[J]．国外药学合成药，生化药制剂分册，1985，6(1)：45-51
22．刘承叶，苗惠珠．儿茶酚胺类注射液的稳定性试验及处方改进[J]．医药工业，1986，17(3)：102-106
23．朱匀芳，刘棠华．应用恒温加速试验法预测药物稳定性的探讨[J]．广东药学，1998，8(2)：1-4
24．何平，王庚保．25％抗坏血酸注射液的稳定性考查及贮存期预测[J]．中国药学杂志，1991，26(6)：349-351
25．郭英喜，黄建忠．400g／l抗坏血酸注射剂变化降解规律及稳定性预测方法的探讨[J]．中国药学杂志，1996，31(4)：214-216
1．王红喜，蒋雪涛．微乳制剂的研究进展[J]．国外医学药学分册，1996，23(4)：206-211
2. Shinoda K, Friberg S. Microemulsions: Colloidal aspects [J]. Advances in Colloid and Interface Science, 1975, 4 (4): 281-300
3. Ingvar Danielsson, Bjorn Lindman. The definition ofa microemulsion [J]. Colloids and Surfaces, 1981, 3:391-392
4．陆彬．纳米乳与亚微乳给药系统[J]．中国药师，2004，7(10)：759-761
5．王晓黎，蒋雪涛．微乳在药剂学上的应用[J]．解放军药学学报，2000，16(2)：88-91
6．王倩，丁红梅，周海源．微乳相图及工艺研究[J]．西北药学杂志，1997，12(2)：72-73
7．张正全，陆彬．微乳给药系统研究概况[J]．中国医药工业杂志，2001，32(3)：139—142
8. Schechter R.S. Microemulsions and Related Systems [M]. New York: Marcel Dekker, 1998, p1-200
9．李干佐，郝京城，李方，刘尚营，汪汉卿．阳离子乳化剂中相微乳液的形成和特性[J]．物理化学 学报，1995，11(6)：553—557
10. Cho Y.W., Flynn M.J. Oral pharmaceutical formulations involving chylomicra formation [P]. WO 9003 164, 1990-04-05
11. Von, C.C., Thoren, P., Engstrom, S. Triglyceride-based microemulsion for intravenous administration of sparingly soluble substances [J]. Journal of Pharmaceutical Science, 1998, 87:200-208
12. Shunmugaperumal Tamilvanan, Sven Schmidt, Rainer H. Müller and Simon Benita. In vitro adsorption of plasma proteins onto the surface (charges) modified-submicron emulsions for intravenous administration [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2005, 59(1): 1-7
13. K.-H. Fro"mming, C. Kraus, W. Mehnert, Physico-chemical properties of the mixed micellar system of Solutol HS15 and sodium deoxycholate [J]. Acta Pharmaceutical. Technology. 1990, 4:214-220
14. T. Reinhart, K.H. Bauer, The hemolysis and solubilization behavior of nonionic polymer surface-active agents classes [J]. Pharmazie,1995, 50:407-408
15．张立超，胡晋红，原永芳，凌代文．非离子乳化剂微乳的制备及抗菌性考察[J]．中国医院药学杂志，2004，24(10)：588-590
16．崔福德．药剂学[M]．北京，中国医药科技出版社，2002，p208
17. Muhannad Jumaa, Bernd W. Mu"ller. Parenteral emulsions stabilized with a mixture of phospholipids and PEG-660-12-hydroxy-stearate: evaluation of accelerated and long-term stability [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 54:207-212
18. M.Y. Levy, S. Benita. Design and characterization of a submicronized o/w emulsion of diazepam for parenteral use [J]. International Journal of Pharmaceutics, 1989, 54:103-112
19. K. Buszello, S. Harnischb, R.H. MuEllerb, B.W. MuEller. The influence of alkali fatty acids on the properties and the stability of parenteral O/W emulsions modified with Solutol HS 15 [J]. European Journal of Pharmaceutics and Biopharmaceutics 2000, 49:143-149
20. Alany RG, Rades T, Agatonovic-Kustrin S, Davies NM, Tucker IG. Effect of alcohols and diols on the phase behaviour of quaternary systems [J]. International Journal of Pharrnaceutics, 2000, 196 (2): 141-145
21．潘国梁，贾晓斌，魏惠华，王勇．药用微乳伪三元相图的几种制备方法比较研究[J]．中国药房，2006，17(1)：9-11
22．鲁莹，蒋雪涛，曾仁杰．卵磷脂微乳的制备与理化性质考察[J]．药学学报，2000，35(1)：52-55
23．鲁莹，吉小欣，高申，刘毅清．利多卡因——卵磷脂微乳处方的初步研究[J]．药学实践杂志，2004，22(3)：141-143
24．俞媛，高申，陈琰，钟延强，程明和．盐酸哌甲酯-卵磷脂微乳的制备及其透皮吸收作用[J]．中国药学杂志，2005，40(11)：843-845
25．陆彬，张正全．用三角相图法研究药用微乳的形成条件[J]．药学学报，2001，36(1)：58-62
26．郭涛，储晓琴，宋洪涛，邓意辉，何进，孙学惠，颜鸣．静脉注射用大蒜油亚微乳的制备[J]．中国药学杂志，2005，40(8)：602-605
27. Seung Rim Hwang, Soo-Jeong Lim, Jeong-Sook Park, Chong-Kook Kim. Phospholipid-based microemulsion formulation of all-trans-retinoic acid for parenteral administration [J]. International Journal of Pharmaceutics, 2004, 276:175-183
28. Kyung-Mi Park a, Mi-Kyung Lee, Ki-Jun Hwang, Chong-Kook Kim. Phospholipid-based microemulsions of flurbiprofen by the spontaneous emulsification process [J]. International Journal of Pharmaceutics, 1999, 183:145-154
29．鲁莹，刘英，蒋雪涛，新型药物载体：微乳[J]．国外医药——合成药 生化药 制剂分册，1999，20(4)：253-258
30. Liang W.P., Tharwat F.T. Novel method to prepare O/W microemlusion within a wide temperature range [J]. Acta Physicochemica Sinica, 2000, 16 (6): 538-542
31．郝京城，邹志陈，鲁绍芬，李文华，柴金玲，李干佐，王汉卿．CTAB微乳液结构研究[J]．化学物理学报，1996，9(6)：547-551
32. D. O. Shah and R. M. Hamlin Jr. Structure of water in microemulsions: electrical, birefringence, and nuclear magnetic resonance studies [J]. Science, 197, 171:483-485
33．王文清，顾国兴．电导法研究水／十二烷基苯磺酸钠／正戊醇／正庚醇体系反向胶束和W／O型微乳液的导电机理[J]．铀冶，1997，16(4)：252-257
34. Lagourette B, Peyrelasse J, Boned C, Clausse, M. Percolative conduction in microemulsion type systems [J]. Nature, 1979, 281:60-62
35．吴文娟，王菊平，杨少湖．十二烷基硫酸钠微乳体系的相行为及电导行为研究[J]．国际医药卫生 导报，2006，12(15)：6-8
36．刁兆玉，刘继金，冷鹏，王泽新．微乳液结构的光散射及折光方法的研究[J]．济南教育学院学报，1999．1：53-55
37．徐信兰，胡晓颖，卢乃弟．纳米乳液的透射电镜样品制备及图像分析[J]．电子显微学报，2002，21(5)：786-787
38. Saito H, Shinoda K. The solubilization of hydrocarbons in aqueous solutions of nonionic surfactants [J]. Journal of Colloid Interface Science, 1967, 24 (1): 10-15
39. Strey R, Jonstromer M. Role of medium-chain alcohols in interracial films of nonionic microemulsions [J]. The Journal of Physical Chemistry. 1992, 96:4537-4542
40. Malcolmson C A. The physicochemical properties of nonionic oil in water microemulsions [M]. Ph D Thesis, University of London, 1992
41．孙瑞元．定量药理学[M]．北京：人民卫生出版社，1987：167—172
42．周海钧．生物检定统计方法[M]．北京，人民卫生出版社，1988：214-216
43．黎七雄，汪晖，肖清秋，孔锐．半数致死量(LD_(50))Bliss法的评价及计算[J]．数理医药学杂志，1995，8(4)：318-320
44．操继跃．生物半数效量通用计算方法的探讨[J]．中国兽药杂志，1992，26(3)：50-53
45．王普民，尹萍．Weibull分布：药物最小致死量的理论分布[J]．沈阳药科大学学报，1997，4(4)：308-310
46．李保华，董向丽，孙显明，赵川德，才秀华，孙立宁．用于计算半数致死量(LD_(50))的SAS程序[J]．莱阳农学院学报，2004，21(4)：269-274
47．章元明．半数致死量LD50的刀切估计及其抽样方差[J]．兽牧兽医学报，1997，28(6)：499—503
1．周长新，邹建凯，陈耀祖，赵昱．气相色谱-质谱法测定当归挥发油中藁本内酯的含量[J]．药物分析杂志，2002，22(4)：290-292
2．林燕芝，唐星，毕开顺．RP-HPLC测定川芎挥发油中藁本内酯的含量[J]．中国中药杂志，2004，29(2)：154-157
3．赵惠茹，周晓棉，冯素香，杨广德，贺浪冲．当归挥发油中藁本内酯在家兔体内的药代动力学[J]．中成药，2005，27(12)：1434—1436
4. Shi YF, He LC, Wang SC. Determination of ligustilide in rat blood and tissues by capillary gas chromatography/mass spectrometry [J]. Biomedical chromatography, 2006, 20 (10): 993-998
5．余奇飞．对凯氏定氮法测定食品中蛋白质含量的测量不确定度评定[J]．农产品加工(学刊)，2006，11：50-51
6．陈凤清，梁忠岩．凯氏定氮法测定松杉灵芝子实体水溶性多糖中蛋白质含量[J]．长春工程学院学报(自然科学版)，2005，6(4)：65-66
7．刘邻渭，陶健，毕磊．双缩脲法测定荞麦蛋白质[J]．食品科学，2004，25(10)：258-261
8．胡卓逸，孙承琦．蛋白质定量方法的进展[J]．生物化学与生物物理进展，1990，17(1)：23-26
9．周卫．Lowry法测定眼宁滴眼液中蛋白质的含量[J]．江苏药学与临床研究，2003，11(1)：20-21
10．张康．生物制品蛋白质含量Lowry测定法应注意的几个问题[J]．海峡药学，2001，13(4)：47-47
11．高敏．分光光度法测定蛋黄提取物中蛋白质的含量[J]．光谱实验室，2003，20(2)：284-286
12．李娟，张耀庭，曾伟，罗璇，廖长春．应用考马斯亮蓝法测定总蛋白含量[J]．中国生物制品学杂志，2000，13(2)：118-120
13．江南，吴开力，黄强，潘苏华．一种简便的考马斯亮蓝G250蛋白质染色方法[J]．生物化学与生物物理进展，2000，27(5)：560-561
14．徐杰伟，温少磊，蔡早育．考马斯亮蓝法测定微量蛋白的条件探讨[J]．江西医学检验，2003，21(5)：353-354
15．童姗姗，余江南，刘文英，安登魁．体内药物分析中的样品预处理技术[J]．药学进展，2001，25(1)：24-27
16．钟大放．以加权最小二乘法建立生物分析标准曲线的若干问题[J]．药物分析杂志，1996，16(5)：343—346
17. Bradford M M. A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding [J]. Analytical Biochemistry, 1976, 72 (1-2): 248-254
18. Ausubel FM, Brent R, Kingston RE, Moore DD. Current protocols in molecular biology [M]. New York: John Willey and Sons Inc, 1994, 10 (1): 1-4
19. M. J. Lee, M. H. Lee, C. K. Shim. Inverse targeting of drugs to eticuloendothelial system-rich organs by lipid microemulsion emulsified with poloxamer 338 [J]. International Journal of Pharmaceutics, 1995, 113:175-187
20. B. B.Lundberg, B. C. Mortimer, T. G. Redgrave. Submicron lipid emulsion containing amphipathic polyethylene glycol for use as drug-carriers with prolonged circulation time [J]. International Journal of Pharmaceutics, 1996, 134:119-127
21. Muhannad Jumaa, Bernd W. Mu"ller. Parenteral emulsions stabilized with a mixture of phospholipids and PEG-660-12-hydroxy-stearate: evaluation of accelerated and long-term stability [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 54:207-212
22. M.Y. Levy, S. Benita. Design and characterization of a submicronized o/w emulsion of diazepam for parenteral use [J]. International Journal of Pharmaceutics, 1989, 54:103-112
23. K. Buszello, S. Harnischb, R.H. MuEllerb, B.W. MuEller. The influence of alkali fatty acids on the properties and the stability of parenteral O/W emulsions modified with Solutol HS 15 [J]. European Journal of Pharmaceutics and Biopharmaceutics 2000, 49:143-149
2．李文志，冯汝就．鸦胆子油乳注射液联合TACE治疗原发性肝癌的临床疗效观察[J]．中药材，2006，29(6)：632-633
3．王云杰，黄立军，张志培，王小平．康莱特注射液治疗原发性肝癌的临床研究[J]．现代肿瘤医学，2006，14(1)：36-37
4．禹玉洪，李雪春，吴涛，屠鹏飞．注射用柴胡挥发油脂质体制备工艺研究[J]．中国中药杂志，2004，29(6)：521-525
5．罗琥捷，李临生．鱼腥草挥发油纳米脂质体的制备[J]．陕西中医，2005，26(12)：1370-1371
6．姚世霞，倪京满，钱松，任凤娇．当归挥发油脂质体制备工艺研究[J]．中成药，2007，29(1)：54-57
7．韩静，唐星，巴德纯．降香挥发油固体脂质纳米粒的制备工艺研究[J]．沈阳药科大学学报，2004， 21 (4): 257-260, 296
8. Malcolmson C, Lawrence M J. A comparison of incorporation of model steroids into non-ionic micellar and microemulsion systems [J]. Journal of Pharmacy and Pharmacology. 1993, 45 (1): 141-143
9. Hoar T P, Schulman J. Transparent water in oil dispersions: oleopathic hydromicelle [J]. Nature (Lond.), 1943, 152:102-103
10. Danielsson I, Lindman B. The definition of a microemulsion [J]. Colloids and Surfaces, 1981, 3: 391-392
11．张正全，陆彬．微乳给药系统研究概况[J]．中国医药工业杂志，2001，32(3)：139-142
12. Winsor P A. Solvent properties of amphiphilic compounds [M]. London, Butterworth's Scientific publication, 1954
13．郭荣，李干佐．阴离子型微乳液的电导行为及其溶液结构[J]．化学学报，1987，45(1)：55-58
14．李泉，李维红，翁诗甫，吴瑾光，徐光宪．水／AOT／正庚烷微乳体系中水结构的FT-IR研究[J]．应用化学，1998，15(1)：1-4
15. Lindman B, Moseley M E, Stilbs P. Fourier-transform NMR self-diffusion and microemulsion structure [J]. Journal of Colloid and Interface Science, 1981, 83 (2): 569-582
16. Jahn W. Srtrey R. Microstructure of microemulsions by freeze fracture electron Microscopy [J]. Journal of Physical Chemistry, 1988, 92:2294-2301
17. Cebula D J, Myers D Y, Ottewill R H. Studies on microemulsions Part 1: Scattering studies on water-in-oil microemulsions [J]. Colloid and Polymer Science, 1982, 260 (1): 96-107
18. Hou M J, Kim M, Shah D O. A light scattering study on the droplet size and interdroplet interaction in microemulsions of AOT-oil-water system [J]. Journal of Colloid Interface Science, 1988, 123 (2): 398-412
19. Attwood D, Ktistic G. A light scattering study on oil-in-water microemulsions [J]. International Journal of Pharmaceutics, 1989, 52 (2): 165-171
20. Shinoda K, Friberg S. Microemulsions: colloidal aspects [J]. Advances in colloid and interfaces science, 1975, 4:281-300
21. Bidner M S, Larson R G, Scriven L E. On the theory of micellization with reference to etroleum recovery [J]. Rev.Latinoam.Ing.Quim.Apl., 1976, 6 (1): 1-32
22．哈润华，侯斯健，栗付平．微乳液结构和丙烯酰胺反相微乳液聚合[J]．高分子通报，1995，1：10-19
23. Prince L M. Microemulsion, theory and practice [M]. Chapter 5, New York: Academic press, 1997.
24. Shinoda K, Araki M, Sadaghiani A, Khan A, Lindman B. Lecithin based microemulsions: phase behavior and microstructure [J]. The Journal of Physical Chemistry, 1991, 95:989-993
25. Aboofazeli R, Lawrence M J. Investigation into the formation and characterization of phosphor-lipids microemulsions. Ⅰ. Pseudo-ternary phase diagrams of systems containing water-lecithin-alcoholisopropylmyristrate [J]. International Journal of Pharmaceutics, 1993, 93: 161-175
26. Milan J S, Katrin S. Microemulsion in technical processes [J]. American Chemical Society, 1995, 95: 849-864
27．吴顺芹，李三鸣，赵国斌．微乳及其在药剂学中的应用[J]．沈阳药科大学学报，2003，20(5)：381-385
28．鲁莹，刘英，蒋雪涛．新型药物载体：微乳[J]．国外医药——合成药 生化药 制剂分册，1999，20(4)：253-258
29．涂秋榕，陈洁，朱家璧．自乳化药物传递系统[J]．国外医药——合成药 生化药 制剂分册，2002，23(3)：170—173
30．崔晶，翟光喜，邹满．微乳给药系统的研究与应用[J]．食品与药品，2005，7(1A)：23-26
31. M. Jayne Lawrence, Gareth D. Rees. Microemulsion based media as novel drug delivery systems [J]. Advanced Drug Delivery Reviews. 2000, 45:89-121
32．王晓黎，蒋雪涛．微乳在药剂学上的应用[J]．解放军药学学报，2000，16(2)：88-91
33．王倩，丁红梅，周海源．微乳相图及工艺研究[J]．西北药学杂志，1997，12(2)：72-73
34．陈华兵，翁婷，杨祥良．微乳在现代药剂学中的研究进展[J]．中国医药工业杂志，2004，35(8)：502-506
35．杨惊宇，严冬，罗英杰，杨大坚，陈士林．新型药物剂型——微乳[J]．中国医学工程，2005，13(4)：378-381
36．陈耀祖，陈能煜，马学毅，李海泉．当归化学成份分析研究——毛细管气相色谱-质谱法鉴定当归根挥发油成份[J]．高等学校化学学报，1984，5(1)：124-128
37. Zhao K J, Dong T, Tu P F, Tsim KWK. Molecular genetic and chemical assessment of Radix angelica (Danggui) in China [J]. Journal of Agricultural and Food Chemistry, 2003, 51: 2576-2583
38．闫升，乔国芳，刘志峰，刘珂，王嘉陵．当归油对大鼠离体子宫平滑肌收缩功能的影响[J]．中草 药，2000，31(8)：604-606
39．刘卫，刘玉玲，吕泰省，刘传绩，俞惠琴．芎归滴丸对海马神经元钠电流的保护作用[J]．中国药学杂志，2002，37(1)：824-826
40．孙所，俞惠琴．芎归滴丸对大鼠脑血流量的影响[J]．山东医药工业，1999，18(4)：44-45
41．郭书英，刘卫，俞惠琴，赵晓民，高允生，苏定冯．芎归滴丸对选择性去窦弓神经大鼠动脉压力感受反射敏感性的影响[J]．中国药房，2003，14(6)：329-331
42. Hou Y Z, Zhao G R, Yang J, Yuan Y J, Zhu G G, Hiltunen R. Protective effect of Ligusticum Chuanxiong and Angelica Sinensis on endothelial cell damage induced by hydrogen peroxide [J]. Life Science, 2004, 75:1775-1786
1．胡长鹰，丁霄霖．当归中藁本内酯的提取、分离与结构鉴定[J]．无锡轻工大学学报，2003，22(5)：69-71
2. Kaouadji M, Pachtere F D, Paouget C, Chulia A J. Three additional phthalide derivatives, an epoxymonomer and two dimers, from Ligusticum wallichii rhizomes [J]. Journal of Natural Products, 1986, 49 (5): 872-877
3．路新华，梁鸿，赵玉英．当归中藁本内酯类化合物的分离与结构鉴定[J]．中国中药杂志，2003，28(5)：423-425
4．宋治中，崔育新，朱启秀，贾忠建．二维H-NMR研究双藁本内酯结构[J]．波谱学杂志，1991，8(2)：209-214
5．宋金春，罗顺德，蔡鸿生，徐锋．反相高效液相色谱法测定生化汤中阿魏酸及其血药浓度[J]．中国医院药学杂志，2001，21(4)：213-215
6．王映芬，何燕萍，蔡阳．复方偏头痛冲剂主要成份当归、川芎中阿魏酸的含量测定[J]．北京医科大学学报，1994，26(3)：218-218
7．贾晓斌，施亚芳，黄一平，上官方．当归和川芎中阿魏酸高效液相色谱测定方法的改进[J]．中成药，1998，20(6)：37-38
8．常明向，刘小平．当归、川芎及配伍后煎煮液中阿魏酸含量的变化[J]．中药饮片，1992，2：24-25
9．梁明金，贺浪冲．藁本内酯和丁烯酜内酯对bFGF诱导的大鼠平滑肌细胞异常增殖的抑制作用[J]．药学学报，2006，41(2)：161-165
10．石力夫，郑晓梅，蔡溱，吴柏生．藁本内酯分解前后川芎挥发油对兔球结膜微循环影响的比较 [J]．中国药理学与毒理学杂志，1995，9(2)：157-158
11．汪程远，杜俊荣，钱忠明．藁本内酯的研究进展[J]．中国药学杂志，2006，41(12)：889-891
12．金灯萍，彭国平，陆晓峰．川芎中藁本内酯对照品的制备[J]．中草药，2006，37(1)：64-65
13．马建龙，李汉蕴，戴荣继，邓玉林．当归化学成分藁本内酯的提取纯化[J]．药品评价，2005，2(3)：214-215
14．苗爱东，梁乾德，刘勇，周喆，王升启．SPE-HPLC制备色谱法从当归挥发油中分离纯化Z-藁本内酯[J]．中药材，2005，28(5)：778-779
15. Li Huabin, Chen Feng. Preparative isolation and purification of chuanxiongzine from the medicinal plant Ligusticum Chuanxiong by high-speed counter-current chromatography [J]. Journal of chromatography A, 2004, 1047:249-253
16．张达磊，李桂生，任召言，曲桂武．气相色谱法测定川芎挥发油中Z-藁本内酯及川芎内酯A的含量[J]．药物分析杂志，2006，26(7)：895-897
17．汪程远，张浩，钱忠明．HPLC测定不同川芎药材中藁本内酯[J]．中草药，2006，37(6)：447-449
18．苗爱东，梁乾德，王升启．SPE-HPLC测定四物合剂中藁本内酯的含量[J]．中药材，2005，27(6)：737-738
19．赵志鸿，刘晨江．薄层荧光扫描法测定参茸近视康复片中藁本内酯的含量[J]．中国实验方剂学杂志，1997，3(2)：4-6
20．王爱武，邱福军，吕文海．双波长薄层扫描法测定当归散中藁本内酯的含量[J]．中国医院药学杂志，2003，23(1)：31-32
21．周长新，邹建凯，陈耀祖，赵昱．气相色谱-质谱法测定当归挥发油中藁本内酯的含量[J]．药物分析杂志，2002，22(4)：290-292
22. Lin Longze, He Xianguo, Lian Lizhi, King Wayne, Elliott Jerry. Liquid chromatographic-electrospray mass spectrometric study of the phthalides of Angelica Sinensis and chemical changes of Z-ligustilide [J]. Journal of Chromatography A, 1998, 810:71-79
23．梁忠明，曹铁城．试述中药提取工艺和新进展[J]．时珍国药研究，1993，4(4)：36-38
24．黄保民．中药提取分离工艺中高新技术应用的进展[J]．中医研究，1998，11(5)：56-58
25．李菁，葛发欢，黄晓芬，李莹，辉国钧．超临界CO_2萃取当归挥发油的研究[J]．中药材，1996，19(4)：187-189
26．胡志方，郭慧玲．川芎、当归挥发油成分混合提取工艺的探讨[J]．江西中医学院学报，2004，16(3)：49-50
27．李桂生，马成俊，刘志锋，刘珂．超临界CO_2萃取法与水蒸气蒸馏法提取当归挥发油的比较[J]．中草药，2001，32(7)：579-583
28．李桂生，刘岩，刘珂．水蒸汽蒸馏法对当归挥发油提取过程中成分异构化的影响[J]．中成药，2001，23(11)：784-786
1．陈耀祖，陈能煜，马学毅，李海泉．当归化学成份分析研究——毛细管气相色谱．质谱法鉴定当归根挥发油成份[J]．高等学校化学学报，1984，5(1)：124-128
2．黄远征，溥发鼎．川芎根茎挥发油化学成分的研究[J]．药学学报，1988，23(6)：426-429
3．石立夫，邓延昭，吴柏生．川芎干燥根茎挥发油化学成分及其稳定性的研究[J]．药物分析杂志，1995，15(3)：26-30
4．李桂生，马成俊，李香玉，刘珂．藁本内酯的稳定性研究及异构化产物的GC-MS分析[J]．中草药，2000，31(6)：405-407
5．周长新，李新华．藁本内酯的稳定性与溶剂化效应的关系[J]．药学学报，2001，36(10)：793-795
6. Bohrmann H, Stahl E, Mitsuhashi H. Constituents of umbelliferous plants (ⅩⅢ). Chromatographic studies on the constituents of cnidium officinale [J]. Chemical and Pharmaceutical Bulletin, 1967, 15:1606-1608
7．李慧，王一涛．藁本内酯稳定性的影响因素及稳定化措施[J]．江西中医学院学报，2003，15(1)：56-57
8．王曦，张曼红，孙杰．硝酸甘油注射液的制备[J]．山东医药工业，1999，18(5)：9-10
9．姜洪志，陆忠贤．苯巴比妥钠注射液贮存期预测[J]．中国药房，1991，2(4)：23-23
10. Naoya Okusa. Prediction of stability of drugs. Ⅲ. Application of Weibull probability paper to prediction of stability [J]. Chemical and Pharmaceutical Bulletin, 1975, 23 (4): 794-802
11．张伟德，叶兴凯．金属离子在液相氧化中的催化作用[J]．分子催化，1993，7(3)：203-211
12. Kobayashi M, Mitsuhashi H. Studies on constituents of Umbelliferae plants. ⅩⅦ. Structures of three new ligustilide derivatives from Ligusticum wallichii [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (12): 4789-4792
13. Kaouadji M, Pouget C, Pouget C. Three additional phthalide derivatives, an epoxymonomer and two dimers, from Ligusticum wallichii rhizomes [J]. Journal of Natural Products. 1986, 49 (5): 872-877
14. Tsuchida T, Kobayashi M, Kaneko K, et al. Studies on the constituents of Umbelliferae plants. ⅩⅥ. Isolation and structures of three new ligustilide derivatives from Angelica acutiloba [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (11): 4460-4464
15. Kobayashi M, Fujita M. Studies on constituent of Umbelliferae plants. ⅩⅤ. Constituents of Cnidium officinale: Occurrence of pregnenolone, coniferylferulate and hydroxyphthalides [J]. Chemical and Pharmaceutical Bulletin, 1987, 35 (4): 1427-1433
16. Lin Longze, He Xianguo, Lian Lizhi, King Wayne, Elliott Herry. Liquid chromatographic electrospray mass spectrometric study of the Angelica sinensis and chemical changes of Z-ligustilide [J]. Journal of Chromatography A, 1998, 810:71-79
17．杜金山．光线与药物的稳定性[J]．天津药学，1995，7(2)：33-35
18．Moore D，郑集声．药物光降解作用原理和实验研究[J]．国外医药药学分册，1998，3：174-177
19．乔传武，东星月，吕秀荣．防止或延缓药物水解保证药物质量的疗效[J]．中国误诊学杂志，2003，3(3)：454-454
20．王仿成．制剂因素与药物质量[J]．安庆医学，2000，21(4)：195—195
21．Mchael JA．药剂产品中的稳定剂[J]．国外药学合成药，生化药制剂分册，1985，6(1)：45-51
22．刘承叶，苗惠珠．儿茶酚胺类注射液的稳定性试验及处方改进[J]．医药工业，1986，17(3)：102-106
23．朱匀芳，刘棠华．应用恒温加速试验法预测药物稳定性的探讨[J]．广东药学，1998，8(2)：1-4
24．何平，王庚保．25％抗坏血酸注射液的稳定性考查及贮存期预测[J]．中国药学杂志，1991，26(6)：349-351
25．郭英喜，黄建忠．400g／l抗坏血酸注射剂变化降解规律及稳定性预测方法的探讨[J]．中国药学杂志，1996，31(4)：214-216
1．王红喜，蒋雪涛．微乳制剂的研究进展[J]．国外医学药学分册，1996，23(4)：206-211
2. Shinoda K, Friberg S. Microemulsions: Colloidal aspects [J]. Advances in Colloid and Interface Science, 1975, 4 (4): 281-300
3. Ingvar Danielsson, Bjorn Lindman. The definition ofa microemulsion [J]. Colloids and Surfaces, 1981, 3:391-392
4．陆彬．纳米乳与亚微乳给药系统[J]．中国药师，2004，7(10)：759-761
5．王晓黎，蒋雪涛．微乳在药剂学上的应用[J]．解放军药学学报，2000，16(2)：88-91
6．王倩，丁红梅，周海源．微乳相图及工艺研究[J]．西北药学杂志，1997，12(2)：72-73
7．张正全，陆彬．微乳给药系统研究概况[J]．中国医药工业杂志，2001，32(3)：139—142
8. Schechter R.S. Microemulsions and Related Systems [M]. New York: Marcel Dekker, 1998, p1-200
9．李干佐，郝京城，李方，刘尚营，汪汉卿．阳离子乳化剂中相微乳液的形成和特性[J]．物理化学 学报，1995，11(6)：553—557
10. Cho Y.W., Flynn M.J. Oral pharmaceutical formulations involving chylomicra formation [P]. WO 9003 164, 1990-04-05
11. Von, C.C., Thoren, P., Engstrom, S. Triglyceride-based microemulsion for intravenous administration of sparingly soluble substances [J]. Journal of Pharmaceutical Science, 1998, 87:200-208
12. Shunmugaperumal Tamilvanan, Sven Schmidt, Rainer H. Müller and Simon Benita. In vitro adsorption of plasma proteins onto the surface (charges) modified-submicron emulsions for intravenous administration [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2005, 59(1): 1-7
13. K.-H. Fro"mming, C. Kraus, W. Mehnert, Physico-chemical properties of the mixed micellar system of Solutol HS15 and sodium deoxycholate [J]. Acta Pharmaceutical. Technology. 1990, 4:214-220
14. T. Reinhart, K.H. Bauer, The hemolysis and solubilization behavior of nonionic polymer surface-active agents classes [J]. Pharmazie,1995, 50:407-408
15．张立超，胡晋红，原永芳，凌代文．非离子乳化剂微乳的制备及抗菌性考察[J]．中国医院药学杂志，2004，24(10)：588-590
16．崔福德．药剂学[M]．北京，中国医药科技出版社，2002，p208
17. Muhannad Jumaa, Bernd W. Mu"ller. Parenteral emulsions stabilized with a mixture of phospholipids and PEG-660-12-hydroxy-stearate: evaluation of accelerated and long-term stability [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 54:207-212
18. M.Y. Levy, S. Benita. Design and characterization of a submicronized o/w emulsion of diazepam for parenteral use [J]. International Journal of Pharmaceutics, 1989, 54:103-112
19. K. Buszello, S. Harnischb, R.H. MuEllerb, B.W. MuEller. The influence of alkali fatty acids on the properties and the stability of parenteral O/W emulsions modified with Solutol HS 15 [J]. European Journal of Pharmaceutics and Biopharmaceutics 2000, 49:143-149
20. Alany RG, Rades T, Agatonovic-Kustrin S, Davies NM, Tucker IG. Effect of alcohols and diols on the phase behaviour of quaternary systems [J]. International Journal of Pharrnaceutics, 2000, 196 (2): 141-145
21．潘国梁，贾晓斌，魏惠华，王勇．药用微乳伪三元相图的几种制备方法比较研究[J]．中国药房，2006，17(1)：9-11
22．鲁莹，蒋雪涛，曾仁杰．卵磷脂微乳的制备与理化性质考察[J]．药学学报，2000，35(1)：52-55
23．鲁莹，吉小欣，高申，刘毅清．利多卡因——卵磷脂微乳处方的初步研究[J]．药学实践杂志，2004，22(3)：141-143
24．俞媛，高申，陈琰，钟延强，程明和．盐酸哌甲酯-卵磷脂微乳的制备及其透皮吸收作用[J]．中国药学杂志，2005，40(11)：843-845
25．陆彬，张正全．用三角相图法研究药用微乳的形成条件[J]．药学学报，2001，36(1)：58-62
26．郭涛，储晓琴，宋洪涛，邓意辉，何进，孙学惠，颜鸣．静脉注射用大蒜油亚微乳的制备[J]．中国药学杂志，2005，40(8)：602-605
27. Seung Rim Hwang, Soo-Jeong Lim, Jeong-Sook Park, Chong-Kook Kim. Phospholipid-based microemulsion formulation of all-trans-retinoic acid for parenteral administration [J]. International Journal of Pharmaceutics, 2004, 276:175-183
28. Kyung-Mi Park a, Mi-Kyung Lee, Ki-Jun Hwang, Chong-Kook Kim. Phospholipid-based microemulsions of flurbiprofen by the spontaneous emulsification process [J]. International Journal of Pharmaceutics, 1999, 183:145-154
29．鲁莹，刘英，蒋雪涛，新型药物载体：微乳[J]．国外医药——合成药 生化药 制剂分册，1999，20(4)：253-258
30. Liang W.P., Tharwat F.T. Novel method to prepare O/W microemlusion within a wide temperature range [J]. Acta Physicochemica Sinica, 2000, 16 (6): 538-542
31．郝京城，邹志陈，鲁绍芬，李文华，柴金玲，李干佐，王汉卿．CTAB微乳液结构研究[J]．化学物理学报，1996，9(6)：547-551
32. D. O. Shah and R. M. Hamlin Jr. Structure of water in microemulsions: electrical, birefringence, and nuclear magnetic resonance studies [J]. Science, 197, 171:483-485
33．王文清，顾国兴．电导法研究水／十二烷基苯磺酸钠／正戊醇／正庚醇体系反向胶束和W／O型微乳液的导电机理[J]．铀冶，1997，16(4)：252-257
34. Lagourette B, Peyrelasse J, Boned C, Clausse, M. Percolative conduction in microemulsion type systems [J]. Nature, 1979, 281:60-62
35．吴文娟，王菊平，杨少湖．十二烷基硫酸钠微乳体系的相行为及电导行为研究[J]．国际医药卫生 导报，2006，12(15)：6-8
36．刁兆玉，刘继金，冷鹏，王泽新．微乳液结构的光散射及折光方法的研究[J]．济南教育学院学报，1999．1：53-55
37．徐信兰，胡晓颖，卢乃弟．纳米乳液的透射电镜样品制备及图像分析[J]．电子显微学报，2002，21(5)：786-787
38. Saito H, Shinoda K. The solubilization of hydrocarbons in aqueous solutions of nonionic surfactants [J]. Journal of Colloid Interface Science, 1967, 24 (1): 10-15
39. Strey R, Jonstromer M. Role of medium-chain alcohols in interracial films of nonionic microemulsions [J]. The Journal of Physical Chemistry. 1992, 96:4537-4542
40. Malcolmson C A. The physicochemical properties of nonionic oil in water microemulsions [M]. Ph D Thesis, University of London, 1992
41．孙瑞元．定量药理学[M]．北京：人民卫生出版社，1987：167—172
42．周海钧．生物检定统计方法[M]．北京，人民卫生出版社，1988：214-216
43．黎七雄，汪晖，肖清秋，孔锐．半数致死量(LD_(50))Bliss法的评价及计算[J]．数理医药学杂志，1995，8(4)：318-320
44．操继跃．生物半数效量通用计算方法的探讨[J]．中国兽药杂志，1992，26(3)：50-53
45．王普民，尹萍．Weibull分布：药物最小致死量的理论分布[J]．沈阳药科大学学报，1997，4(4)：308-310
46．李保华，董向丽，孙显明，赵川德，才秀华，孙立宁．用于计算半数致死量(LD_(50))的SAS程序[J]．莱阳农学院学报，2004，21(4)：269-274
47．章元明．半数致死量LD50的刀切估计及其抽样方差[J]．兽牧兽医学报，1997，28(6)：499—503
1．周长新，邹建凯，陈耀祖，赵昱．气相色谱-质谱法测定当归挥发油中藁本内酯的含量[J]．药物分析杂志，2002，22(4)：290-292
2．林燕芝，唐星，毕开顺．RP-HPLC测定川芎挥发油中藁本内酯的含量[J]．中国中药杂志，2004，29(2)：154-157
3．赵惠茹，周晓棉，冯素香，杨广德，贺浪冲．当归挥发油中藁本内酯在家兔体内的药代动力学[J]．中成药，2005，27(12)：1434—1436
4. Shi YF, He LC, Wang SC. Determination of ligustilide in rat blood and tissues by capillary gas chromatography/mass spectrometry [J]. Biomedical chromatography, 2006, 20 (10): 993-998
5．余奇飞．对凯氏定氮法测定食品中蛋白质含量的测量不确定度评定[J]．农产品加工(学刊)，2006，11：50-51
6．陈凤清，梁忠岩．凯氏定氮法测定松杉灵芝子实体水溶性多糖中蛋白质含量[J]．长春工程学院学报(自然科学版)，2005，6(4)：65-66
7．刘邻渭，陶健，毕磊．双缩脲法测定荞麦蛋白质[J]．食品科学，2004，25(10)：258-261
8．胡卓逸，孙承琦．蛋白质定量方法的进展[J]．生物化学与生物物理进展，1990，17(1)：23-26
9．周卫．Lowry法测定眼宁滴眼液中蛋白质的含量[J]．江苏药学与临床研究，2003，11(1)：20-21
10．张康．生物制品蛋白质含量Lowry测定法应注意的几个问题[J]．海峡药学，2001，13(4)：47-47
11．高敏．分光光度法测定蛋黄提取物中蛋白质的含量[J]．光谱实验室，2003，20(2)：284-286
12．李娟，张耀庭，曾伟，罗璇，廖长春．应用考马斯亮蓝法测定总蛋白含量[J]．中国生物制品学杂志，2000，13(2)：118-120
13．江南，吴开力，黄强，潘苏华．一种简便的考马斯亮蓝G250蛋白质染色方法[J]．生物化学与生物物理进展，2000，27(5)：560-561
14．徐杰伟，温少磊，蔡早育．考马斯亮蓝法测定微量蛋白的条件探讨[J]．江西医学检验，2003，21(5)：353-354
15．童姗姗，余江南，刘文英，安登魁．体内药物分析中的样品预处理技术[J]．药学进展，2001，25(1)：24-27
16．钟大放．以加权最小二乘法建立生物分析标准曲线的若干问题[J]．药物分析杂志，1996，16(5)：343—346
17. Bradford M M. A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding [J]. Analytical Biochemistry, 1976, 72 (1-2): 248-254
18. Ausubel FM, Brent R, Kingston RE, Moore DD. Current protocols in molecular biology [M]. New York: John Willey and Sons Inc, 1994, 10 (1): 1-4
19. M. J. Lee, M. H. Lee, C. K. Shim. Inverse targeting of drugs to eticuloendothelial system-rich organs by lipid microemulsion emulsified with poloxamer 338 [J]. International Journal of Pharmaceutics, 1995, 113:175-187
20. B. B.Lundberg, B. C. Mortimer, T. G. Redgrave. Submicron lipid emulsion containing amphipathic polyethylene glycol for use as drug-carriers with prolonged circulation time [J]. International Journal of Pharmaceutics, 1996, 134:119-127
21. Muhannad Jumaa, Bernd W. Mu"ller. Parenteral emulsions stabilized with a mixture of phospholipids and PEG-660-12-hydroxy-stearate: evaluation of accelerated and long-term stability [J]. European Journal of Pharmaceutics and Biopharmaceutics, 2002, 54:207-212
22. M.Y. Levy, S. Benita. Design and characterization of a submicronized o/w emulsion of diazepam for parenteral use [J]. International Journal of Pharmaceutics, 1989, 54:103-112
23. K. Buszello, S. Harnischb, R.H. MuEllerb, B.W. MuEller. The influence of alkali fatty acids on the properties and the stability of parenteral O/W emulsions modified with Solutol HS 15 [J]. European Journal of Pharmaceutics and Biopharmaceutics 2000, 49:143-149
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