反溶剂法制备叶黄素酯纳米粒
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摘要
目的优化叶黄素酯(LE)纳米粒(NPs)的制备工艺,并对其水溶性的改善程度进行测试。方法以四氢呋喃为溶剂,去离子水为反溶剂,泊洛沙姆188作为表面活性剂,通过反溶剂法制备LE-NPs,采用单因素实验设计法优化LE-NPs制备工艺。并综合利用扫描电子显微镜、激光粒度仪、X射线衍射、差示扫描量热法等分析方法对获得的LE-NPs理化性质进行表征;采用气相色谱(GC)进行溶剂残留测定;并进行了体外溶出对比实验。结果 LE-NPs的最优制备条件为沉积时间10 min,叶黄素酯质量浓度50 mg/mL,溶剂反溶剂体积比为1∶7,泊洛沙姆188质量分数0.5%,搅拌速率950 r/min,沉积温度25℃;所得LE-NPs为球形,平均粒径为164 nm,其中四氢呋喃残留量为344.3μg/g,低于人用药物注册技术要求国际协调会(ICH)要求的最低标准(0.780 mg/g);在人工胃液环境中LE-NPs的饱和溶解度和溶出速率分别是原药的2.91倍和9.65倍。结论反溶剂法制备的LE-NPs具有较好水溶性,在口服制剂中具有潜在的用途。
Objective To prepare lutein ester(LE) nanoparticles(LE-NPs) by anti-solvent method using tetrahydrofuran as solvent, deionized water as antisolvent and Poloxamer 188 as surfactant, and to optimize the process of getting LE-NPs. Methods The concentration of LE, ratio of solvent to solvent, type and dosage of surfactant, precipitation time and stirring speed were investigated factors affecting the particle size of LE-NPs. The raw LE and LE-NPs were observed and analyzed by scanning electron microscopy, laser particle size analyzer, X-ray diffraction, and differential scanning calorimetry; Solvent residue was tested by GC method. What's more, the solubility and dissolving capability of raw LE and LE-NPs also were detected in vitro in this study. Results The optimized preparation conditions of LE-NPs were as follows: 10 min of stirring time, 50 mg/mL of LE concentration, 1∶7 of the volume ratio of solvent to antisolvent, 0.5% at mass fraction of poloxamer 188, 950 r/min of stirring speed, 25 ℃ of precipitation temperature. The mean particle size of spherical LE-NPs was 164 nm. XRD and DSC results showed that LE-NPs had lower crystallinity compared to raw drug, and mainly in amorphous state. The solvent residue result showed that tetrahydrofuran residue content in LE-NPs was 344.3 μg/g. Furthermore, the solubility and dissolution rate of LE-NPs were about 2.91 times and 9.65 times of raw LE. Conclusion LE-NPs prepared by antisolvent method could become a new formulation, which has higher solubility and bioavailability than raw LE.
Objective To prepare lutein ester(LE) nanoparticles(LE-NPs) by anti-solvent method using tetrahydrofuran as solvent, deionized water as antisolvent and Poloxamer 188 as surfactant, and to optimize the process of getting LE-NPs. Methods The concentration of LE, ratio of solvent to solvent, type and dosage of surfactant, precipitation time and stirring speed were investigated factors affecting the particle size of LE-NPs. The raw LE and LE-NPs were observed and analyzed by scanning electron microscopy, laser particle size analyzer, X-ray diffraction, and differential scanning calorimetry; Solvent residue was tested by GC method. What's more, the solubility and dissolving capability of raw LE and LE-NPs also were detected in vitro in this study. Results The optimized preparation conditions of LE-NPs were as follows: 10 min of stirring time, 50 mg/mL of LE concentration, 1∶7 of the volume ratio of solvent to antisolvent, 0.5% at mass fraction of poloxamer 188, 950 r/min of stirring speed, 25 ℃ of precipitation temperature. The mean particle size of spherical LE-NPs was 164 nm. XRD and DSC results showed that LE-NPs had lower crystallinity compared to raw drug, and mainly in amorphous state. The solvent residue result showed that tetrahydrofuran residue content in LE-NPs was 344.3 μg/g. Furthermore, the solubility and dissolution rate of LE-NPs were about 2.91 times and 9.65 times of raw LE. Conclusion LE-NPs prepared by antisolvent method could become a new formulation, which has higher solubility and bioavailability than raw LE.
引文
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[20]赵修华,刘影,王卫国,等.微粉化反式肉桂酸的反溶剂重结晶法制备与表征[J].中草药,2014,45(15):2165-2171.
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[2]陈龙胜,忻旸,周斌.叶黄素酯的生物转化研究[J].中草药,2005,36(10):1486-1488.
[3]张秀娟,安鹏,季宇彬.叶黄素酯对四氧嘧啶所致小鼠氧化损伤的保护性研究[J].中国微生态学杂志,2008,20(1):25-27.
[4]孟祥河,毛忠贵,潘秋月.叶黄素的保健功能[J].中国食品添加剂,2003,10(1):17-20.
[5]惠伯棣,张凌霄,张艳.叶黄素酯在体内的相对生物利用度评价[J].食品科学,2009,30(3):252-255.
[6]孙震,姚惠源.叶黄素的抗癌作用及其研究现状[J].生物技术通讯,2005,16(1):84-86.
[7]李海毅.叶黄素的质量标准研究及抗氧化活性评价[D].长春:吉林大学,2012.
[8]黄媛,柴晓磊,陈碧聪,等.叶黄素酯CWS微囊粉理化性质与稳定性能研究[J].中国食品添加剂,2012,19(4):71-74.
[9]姜鹏飞,程惠,韩阳,等.叶黄素酯肠溶微囊的制备工艺研究及性能考察[J].中国食品添加剂,2013(4):125-129.
[10]Amar I,Aserin A,Garti N.Solubilization patterns of lutein and lutein esters in food grade nonionic microemulsions[J].J Agric Food Chem,2003,51(16):4775-4781.
[11]岳鹏飞,但济修,谢元彪,等.难溶性中药成分纳米晶体给药系统的研究进展与思考[J].世界中医药,2015,10(3):310-314.
[12]谢元彪,许俊男,陈颖翀,等.纳米晶体技术在难溶性药物中的应用进展与思考[J].世界科学技术—中医药现代化,2016,18(10):1788-1793.
[13]葛云龙,赵修华,祖元刚,等.姜黄素纳米粒冻干粉的反溶剂法制备工艺优化及溶出特征[J].中草药,2016,47(14):2447-2453.
[14]张艳,惠伯棣,张凌霄.叶黄素酯在体内消化吸收过程中水解的研究[J].食品科学,2007,28(8):461-465.
[15]李大婧,方桂珍,刘春泉,等.叶黄素酯和叶黄素稳定性的研究[J].林产化学与工业,2007,27(1):112-116.
[16]许文佳,赵修华,祖元刚,等.反溶剂法制备甘草酸纳米粒的表征与体外透皮特性研究[J].中草药,2014,45(8):1068-1071.
[17]Zhang J Y,Shen Z G,Zhong J,et al.Preparation of amorphous cefuroxime axetil nanoparticles by controlled nanoprecipitation method without surfactants[J].Int J Pharm,2006,323(1/2):153-160.
[18]Zhe W,Chen J F,Le Y,et al.Preparation of ultrafine beclomethasone dipropionate drug powder by antisolvent precipitation[J].Ind Eng Chem Res,2007,46(14):4839-4845.
[19]祖元刚,赵野,赵修华,等.响应面设计法优化反溶剂重结晶制备甘草酸微粉[J].森林工程,2011,27(3):54-60.
[20]赵修华,刘影,王卫国,等.微粉化反式肉桂酸的反溶剂重结晶法制备与表征[J].中草药,2014,45(15):2165-2171.
[21]Li X S,Wang J X,Shen Z G,et al.Preparation of uniform prednisolone microcrystals by a controlled microprecipitation method[J].Int J Pharm,2007,342(2):26-32.
[22]Zhao H,Wang J X,Wang Q A,et al.Controlled liquid antisolvent precipitation of hydrophobic pharmaceutical nanoparticles in a microchannel reactor[J].Ind EngChem Res,2007,46(24):8229-8235.
[23]谢玉洁,乐园,王洁欣,等.反溶剂重结晶法制备青蒿素超细粉体[J].化工学报,2012,63(5):1607-1614.
[24]Dokoumetzidis A,Papadopoulou V,Macheras P.Analysis of dissolution data using modified versions of Noyes-Whitney equation and the Weibull function[J].Pharm Res,2006,23(2):256-261.
[25]付骋宇,李莹,梁靓,等.快速甲酯化-气相色谱法测试油脂组成[J].中国粮油学报,2014,29(6):104-107.