CN104549162A - 一种用于分离手性物质的磁场流化床固定相的制备方法 - Google Patents

一种用于分离手性物质的磁场流化床固定相的制备方法 Download PDF

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CN104549162A
CN104549162A CN201410614340.3A CN201410614340A CN104549162A CN 104549162 A CN104549162 A CN 104549162A CN 201410614340 A CN201410614340 A CN 201410614340A CN 104549162 A CN104549162 A CN 104549162A
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华文蔚
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Abstract

一种用于分离手性物质的磁场流化床固定相的制备方法：包括(1)、将三氯化铁溶于稀盐酸水溶液中，滴加氨水至有絮状黑色沉淀出现，过滤，洗涤，蒸发制得磁性纳米颗粒；(2)、将磁性纳米颗粒的水分散液加入到醇的水溶液中混合，加入氢氧化钠溶液和正硅酸烷基酯，得到二氧化硅包裹的磁性颗粒；(3)、在步骤(2)得到的磁性颗粒清洗干燥后加入硅烷偶联剂；(4)、收集步骤(3)所得的产物，然后分别用乙醇和水洗涤产物，最后将产物分散在去离子水中，得到经硅烷偶联剂改性的磁性纳米颗粒的水分散液；(5)、将异丙基丙烯酰胺、环糊精或牛血清蛋白、高分子交联剂和引发剂溶于由步骤(4)所制得磁性纳米颗粒分散液中，通氮气并搅拌。

Description

一种用于分离手性物质的磁场流化床固定相的制备方法
技术领域
本发明涉及一种生物医药领域中用于分离手性物质的磁场流化床固定相的制备方法。
背景技术
超顺磁性纳米/微米颗粒由于具有很大的表面积，表面活性很高，因此，在热力学上很不稳定，很容易凝聚成团。为了防止磁性颗粒之间的凝聚作用，使其形成稳定的胶体溶液，需要在制备过程中对磁性颗粒表面进行修饰，经过修饰的磁性颗粒表面应带有较丰富的活性功能基团，如-NH2、-COOH、-OH和-CHO等，以便偶联生物分子和亲和配基，得到适合各种应用的适合磁性载体。磁性颗粒内部含有磁性粒子，具有超顺磁性，当在有外加磁场下表现出较好的磁性，而在去除外加磁场后，磁性颗粒的剩磁为零，因而其可在外加磁场的作用下方便地控制和分离。此磁性颗粒表面本身具有或通过表面改性带有多种活性的功能基团，可进行生物活性物质(如蛋白质、核酸、酶等)的分离，可以偶联特异性分子(如特异性配体、抗体、抗原等)来专一性地分离生物大分子，也可进行酶的固定化并进行相应的酶催化反应。此外，超顺磁性纳米/微米颗粒在经过表面修饰后，可直接用于化学合成、石油化工等领域的化学反应催化剂进行化学催化反应。因而，超顺磁性纳米/微米载体在生物产品的分离、酶催化、化学催化等领域有着广泛的应用前景。
磁场流化床(Magnetically Fluidized Bed，MFB)是将外加的磁场引入普通的流化床，采用磁敏性颗粒作为床层介质的固液相处理系统，是流态化技术与电磁技术相结合的产物，是一种新型、高效的流态化技术。而磁稳定流化床(Magnetically Stabilized Fluidized Bed，MSFB)是MFB的特殊形式，它是在轴向不随时间变化的均匀磁场下形成的、只有微弱运动的稳定床层。MFB和MSFB都具有振动小、噪声小、稳定性好、操作范围宽和适应性广等特点，同时集固定床与传统流化床的优点于一身：固体颗粒装卸方便、可循环使用；传质传热速率高等，有很高的开发价值和广阔的应用前景。
为了满足超顺磁性纳米/微米颗粒在生物产品的分离、酶催化、化学催化等领域的应用的需求，磁稳定流化床被得以广泛的应用。然而，随着材料科学的发展，采用的磁性颗粒都是具有较大表面积的纳米级或微米级颗粒，这导致磁性颗粒极易从磁场流化床中逸出。磁稳定流化床一般采用电磁线圈来产生磁场，在磁稳定流化床的实际应用过程中存在着下述问题：少量细磁性颗粒载体/催化剂从反应器出口处被液体夹带流出，造成分离介质的流失；为防流失，通常通过增加外加电磁线圈的电压和电流来加大磁场强度，这一做法不但会增加磁场流化床操作的能耗，而且在强磁场下，磁稳定流化床处于冻结区，磁性吸附载体呈链状，床内出现沟流现象，流体分布不均匀，导致传质效率降低。
随着生命科学的进步和制药工业的发展，人类对手性异构在生命体中所起的作用认识越来越深刻。例如：互为对映体的药物在人体内的药理往往是不同的，有的甚至相反，在处理有生物活性的物质，如：维生素、信息素、辅醇、药物产品及一般天然存在的手性化合物时，对映体的分离和分析都是头等重要的问题，因此对映体的手性拆分日益受到人们的重视。就药学领域来说，就有多达30％～40％的药物具有手性。手性异构体在生物活性或药物动力学方面的差异使得手性对映体的分离尤为重要。震惊世界的沙立度胺致畸事件就是一个忽视立体化学效应的恶行事故。基于对映体分子的光学性质与其生物活性之间的特殊相关性，在1992年美国食品药品管理局(FAD)就做出了规定，凡研制具有不对称中心的药物，在药物的鉴定和审批报告中必须给出手性拆分结果。相应地，欧共体国家也提出了相类似的措施。另外，在环境化学研究领域，近年的研究也发现，一些手性异构体的环境效应和生态毒理学效应也不尽相同。因此，建立和发展快速准确的手性对映体拆分方法对于分析化学研究具有相当重要的实际意义。
众所周知，要实现分子式相同，化学性质及其相似的手性对映体的拆分必须有手性拆分试剂的参与。现在的手性对映体拆分方法中，主要有两个方面：把手性拆分试剂添加到流动相中，手性识别剂随着分离的进行而流走损失；手性拆分试剂固定于分离分析的固定相中，在一段时间内手性识别剂的分子识别效率保持不变。后一种手性识别剂的手性识别形式由于手性识别剂能够得到反复利用、分离重现性好、分离效率高而受到广大分析工作者的关注。
目前，对手性分离的困难也存在难以扩大生产规模的问题，将实验室使用的分离柱放大到工业规模中无法使用，固定性颗粒小造成了压力过大，效率过低，而且固定相溶液失活的问题；为了解决上述问题，本发明利用了磁场作为外力而使固定相在分离柱中形成稳定的流态化，解决了上述问题，可以适用于工业化的生产规模。
发明内容
本发明的目的在于提出一种用于分离手性物质的磁场流化床固定相的制备方法。
为达此目的，本发明采用以下技术方案：
一种用于分离手性物质的磁场流化床固定相的制备方法：包括如下步骤：
(1)、将三氯化铁溶于稀盐酸水溶液中，配制成浓度为1-10％重量的溶液，缓慢滴加浓度为1％-10％的氨水至有絮状黑色沉淀出现，直到PH为10～12，静置24-48小时，过滤，洗涤至PH＝7，将纳米粒子移至无水乙醇中，所得悬浮液在40-90℃下搅拌蒸发制得磁性纳米颗粒；
(2)、将一定量的磁性纳米颗粒的水分散液加入到醇的水溶液中混合，在搅拌下加入氢氧化钠溶液和正硅酸烷基酯，反应温度为10-30℃，反应时间12-36小时；磁性纳米颗粒的加入量占整个体系重量的0.5-1.0％，醇占50-65％，碱溶液占1.0-2.0％，正硅酸烷基酯占0.5-1.0％；得到二氧化硅包裹的磁性颗粒；
(3)、在步骤(2)得到的二氧化硅包裹的磁性颗粒经过清洗干燥后加入硅烷偶联剂，反应12-24小时；
(4)、通过离心分离收集步骤(3)所得的产物，然后分别用乙醇和水洗涤产物，最后将产物分散在去离子水中，得到经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒的水分散液，其浓度为0.5-5.0wt％；
(5)、将异丙基丙烯酰胺、环糊精或牛血清蛋白、高分子交联剂和引发剂溶于由步骤(4)所制得的经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒分散液中，通氮气并搅拌10-60分钟后，将整个体系置于60-80℃，反应2-6小时；整个体系中按重量计算，二氧化硅包裹的的磁性纳米颗粒占0.5-1.5％，异丙基丙烯酰胺0.5-4.0％，环糊精或牛血清蛋白重量占总重量的1-10％，引发剂过硫酸钾占总重量的0.1-1％。
具体实施方式
实施例1
(1)、将三氯化铁溶于稀盐酸水溶液中，配制成浓度为1％重量的溶液，缓慢滴加浓度为10％的氨水至有絮状黑色沉淀出现，直到PH为12，静置48小时，过滤，洗涤至PH＝7，将纳米粒子移至无水乙醇中，所得悬浮液在90℃下搅拌蒸发制得磁性纳米颗粒；
(2)、将一定量的磁性纳米颗粒的水分散液加入到醇的水溶液中混合，在搅拌下加入氢氧化钠溶液和正硅酸乙酯，反应温度为30℃，反应时间36小时；磁性纳米颗粒的加入量占整个体系重量的1.0％，醇占65％，碱溶液占2.0％，正硅酸烷基酯占1.0％；得到二氧化硅包裹的磁性颗粒；
(3)、在步骤(2)得到的二氧化硅包裹的磁性颗粒经过清洗干燥后加入硅烷偶联剂，反应24小时；
(4)、通过离心分离收集步骤(3)所得的产物，然后分别用乙醇和水洗涤产物，最后将产物分散在去离子水中，得到经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒的水分散液，其浓度为5.0wt％；
(5)、将异丙基丙烯酰胺、环糊精或牛血清蛋白、高分子交联剂和引发剂溶于由步骤(4)所制得的经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒分散液中，通氮气并搅拌60分钟后，将整个体系置于80℃，反应6小时；整个体系中按重量计算，二氧化硅包裹的的磁性纳米颗粒占1.5％，异丙基丙烯酰胺4.0％，环糊精或牛血清蛋白重量占总重量的10％，引发剂过硫酸钾占总重量的1％。
实施例2
(1)、将三氯化铁溶于稀盐酸水溶液中，配制成浓度为10％重量的溶液，缓慢滴加浓度为1％-％的氨水至有絮状黑色沉淀出现，直到PH为10，静置48小时，过滤，洗涤至PH＝7，将纳米粒子移至无水乙醇中，所得悬浮液在90℃下搅拌蒸发制得磁性纳米颗粒；
(2)、将一定量的磁性纳米颗粒的水分散液加入到醇的水溶液中混合，在搅拌下加入氢氧化钠溶液和正硅酸丁酯，反应温度为30℃，反应时间36小时；磁性纳米颗粒的加入量占整个体系重量的1.0％，醇占65％，碱溶液占2.0％，正硅酸烷基酯占1.0％；得到二氧化硅包裹的磁性颗粒；
(3)、在步骤(2)得到的二氧化硅包裹的磁性颗粒经过清洗干燥后加入硅烷偶联剂，反应24小时；
(4)、通过离心分离收集步骤(3)所得的产物，然后分别用乙醇和水洗涤产物，最后将产物分散在去离子水中，得到经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒的水分散液，其浓度为5.0wt％；
(5)、将异丙基丙烯酰胺、环糊精或牛血清蛋白、高分子交联剂和引发剂溶于由步骤(4)所制得的经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒分散液中，通氮气并搅拌10分钟后，将整个体系置于60℃，反应2小时；整个体系中按重量计算，二氧化硅包裹的的磁性纳米颗粒占1.5％，异丙基丙烯酰胺4.0％，环糊精或牛血清蛋白重量占总重量的10％，引发剂过硫酸钾占总重量的1％。

Claims (1)

1.一种用于分离手性物质的磁场流化床固定相的制备方法：其特征在于包括如下步骤：
(1)、将三氯化铁溶于稀盐酸水溶液中，配制成浓度为1-10％重量的溶液，缓慢滴加浓度为1％-10％的氨水至有絮状黑色沉淀出现，直到PH为10～12，静置24-48小时，过滤，洗涤至PH＝7，将纳米粒子移至无水乙醇中，所得悬浮液在40-90℃下搅拌蒸发制得磁性纳米颗粒；
(2)、将一定量的磁性纳米颗粒的水分散液加入到醇的水溶液中混合，在搅拌下加入氢氧化钠溶液和正硅酸烷基酯，反应温度为10-30℃，反应时间12-36小时；磁性纳米颗粒的加入量占整个体系重量的0.5-1.0％，醇占50-65％，碱溶液占1.0-2.0％，正硅酸烷基酯占0.5-1.0％；得到二氧化硅包裹的磁性颗粒；
(3)、在步骤(2)得到的二氧化硅包裹的磁性颗粒经过清洗干燥后加入硅烷偶联剂，反应12-24小时；
(4)、通过离心分离收集步骤(3)所得的产物，然后分别用乙醇和水洗涤产物，最后将产物分散在去离子水中，得到经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒的水分散液，其浓度为0.5-5.0wt％；
(5)、将异丙基丙烯酰胺、环糊精或牛血清蛋白、高分子交联剂和引发剂溶于由步骤(4)所制得的经硅烷偶联剂改性的二氧化硅包裹的磁性纳米颗粒分散液中，通氮气并搅拌10-60分钟后，将整个体系置于60-80℃，反应2-6小时；整个体系中按重量计算，二氧化硅包裹的的磁性纳米颗粒占0.5-1.5％，异丙基丙烯酰胺0.5-4.0％，环糊精或牛血清蛋白重量占总重量的1-10％，引发剂过硫酸钾占总重量的0.1-1％。
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