CN114230718B - 用于吸附血液中低密度脂蛋白的树脂的制备方法及应用 - Google Patents
用于吸附血液中低密度脂蛋白的树脂的制备方法及应用 Download PDFInfo
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Abstract
本发明公开了用于吸附血液中低密度脂蛋白的树脂的制备方法及应用，属于体外血液净化的吸附树脂技术领域；包含以下步骤：(1)配置分散相；(2)配置聚合相；(3)反向悬浮聚合。本发明以亲水性单体为聚合物，疏水性环己烷为分散相，海藻酸钠和丙烯酸能够提供大量的羧酸基团，聚乙烯醇能够提供大量的羟基，通过这些功能基团上所带的负电荷吸附带正电荷的脂蛋白，来达到固定吸附脂蛋白的目的。
Description
技术领域
本发明涉及领域属于体外血液净化的吸附树脂技术领域，尤其是涉及用于吸附血液中低密度脂蛋白的树脂的制备方法及应用。
背景技术
人体血液中的脂类以脂蛋白的形式存在,可分为高密度脂蛋白(HDL)，低密度脂蛋白(LDL)，极低密度脂蛋白(VDL)，乳糜微粒(CM)，血浆中某种或某几种脂蛋白的浓度超过正常水平称之为高脂蛋白血症，此病症通常伴有血浆脂质升高。
VLDL及LDL是富含胆固醇及胆固醇脂的脂蛋白，在正常情况下，它们可以通过分解代谢作用排出体外。医学研究结果表明，血管硬化及心血管病患者的血液中LDL及VLDL的含量远高于正常值，高浓度的LDL和VLDL是上述疾病的致病因素之一。一般患者可通过饮食及药物疗法降低LDL及VLDL的含量而达到治疗的目的，对于家族性遗传性高脂血症患者，由于其体内缺乏LDL及VLDL受体，因此LDL及VLDL不能通过正常的分解代谢作用排出体外而危及患者的生命。体外血液净化的方法(如：选择性吸附或滤过)可以清除这类患者血液中高浓度的LDL及VLDL，达到挽救患者的目，根据各种脂蛋白表面所带的电性、疏水性及颗粒大小等性质的不同可以设计对LDL具有高吸附选择性的吸附剂，如将肝素或磺化葡聚糖等聚阴离子固载到各种载体而制成聚阴离子吸附剂，前者的灌流过程中需要超生理浓度的Ca2+存在，后者具有良好的吸附选择性，但存在配基的价格昂贵的缺憾；将抗LDL抗体或特定的酶偶联到载体表面可得到免疫吸附剂，它们具有优良的吸附选择性和良好的血液相容性，同时也存在配基不易得到，价格昂贵及消毒困难等缺点。
因此急需开发一种价格低廉、良好的血液相容性、制备过程简单、安全、有效的LDL吸附剂。
发明内容
为了解决上述问题，本发明使用反向悬浮聚合的方法，以亲水性单体为聚合物，疏水性环己烷为分散相，通过海藻酸钠和丙烯酸提供大量的羧酸基团，聚乙烯醇提供大量的羟基，通过这些功能基团上所带的负电荷吸附带正电荷的载脂蛋白已达到固定吸附脂蛋白的目的，本发明所采取的技术方案为：
1、用于吸附血液中低密度脂蛋白的树脂的制备方法及应用，包含以下步骤：
1)配置分散相：在反应容器中加入环己烷和分散剂，温度35-50℃，搅拌25-35min，制得分散相备用；
2)配置聚合相：在反应容器中加入海藻酸钠、甲基丙烯酸、聚乙烯醇水溶液和戊二醛水溶液，搅拌至完全溶解，制得聚合相备用；
3)反向悬浮聚合：将配制好的聚合相缓慢加入分散相，搅拌至聚合相分散为均匀的珠粒，温度升至58-62℃，向聚合体系加0.1-0.3mol/L盐酸水溶液，珠粒定型后将温度升至70-78℃，再加入0.1-0.3mol/LNaOH水溶液，反应2-4h后，过滤出树脂，用95％乙醇洗涤至洗脱液清澈，用0.2mol/L碱溶液浸泡6-10h，再用蒸馏水洗至中性，用乙醇回流提取24h，真空干燥。
所述分散剂占分散相的质量百分比为2％-5％。
所述步骤(2)中的聚合相由以下重量百分比的各组分组成：海藻酸钠25-35％，甲基丙烯酸30-50％，聚乙烯醇水溶液15-25％，戊二醛水溶溶液8-15％。
所述聚乙烯醇水溶液的质量分数为20-40％，戊二醛水溶溶液的质量分数为50-70％。
所述分散剂为SPan-80。
所述分散相和聚合相的质量比为2-5:1。
2、本发明的树脂在吸附低密度脂蛋白中的应用。
本发明与现有技术相比，具有以下优点：
1、本发明采用反向悬浮聚合的方式，以亲水性聚合单体进行多元聚合，在大量的亲水基团存在下能够提高吸附量，另一方面这些亲水基团同时也能够提高树脂的生物相容性。
2、海藻酸钠和丙烯酸能够提供大量的羧酸基团，聚乙烯醇能够提供大量的羟基，通过这些功能基团上所带的负电荷吸附带正电荷的载脂蛋白已达到固定吸附脂蛋白的目的。
3、本发明的树脂对低密度脂蛋白吸附，最高可达89％以上。
附图说明
图1为实施例1合成的树脂的图片。
具体实施方式
以下通过具体实施例进一步说明本发明，但本领域技术人员应当知晓本发明的具体实施例并不以任何方式限制本发明，且在本发明基础上所作出的任何等同替换均落入本发明的保护范围。
实施例1：
1、合成树脂
在装有搅拌器、温度计、回流冷凝管的三口瓶中，加入870g环己烷、30g分散剂SPan-80，升温45℃搅拌30min，得到分散相；将90g海藻酸钠，120g甲基丙烯酸，30％w/w聚乙烯醇水溶液60g，60％w/w戊二醛水溶液30g，分别加入烧杯中常温搅拌10min配制成聚合相，分散相与聚合相之比为3：1，把玻璃棒插入三口瓶底部，将聚合相沿着玻璃棒加入分散相，调节搅拌速度，待聚合相分散为均匀珠粒，温度升至58-62℃，向聚合体系加0.2mol/L盐酸水溶液20ml，珠粒定型后将温度升至70-78℃，再加入0.2mol/L NaOH水溶液20ml反应3h，反应完毕过滤出树脂用95％乙醇洗涤至洗脱液清澈，用0.2mol/L碱溶液浸泡8小时，再用蒸馏水洗到中性，用乙醇回流提取24h，真空干燥。
2、测定树脂白球理化指标(见表1)
水分：用纯化水浸泡树脂24小时，之后离心，使用快速水分仪测试树脂白球水分。
粒度：使用不同孔径的筛子进行测量。
功能基含量：使用赛默飞红外光谱仪测量。
平均孔径和比表面积：使用BET孔径比表面积分析仪测量。
表1实施例1合成的树脂的理化指标
从表1可以看出，树脂成功的接枝了羧酸和羟基，可以有效的吸附脂蛋白，本身树脂未进行制孔，现有的孔结构是树脂吸水后形成的孔结构，树脂主要以表面吸附为主。
实施例2：
1、合成树脂
在装有搅拌器、温度计、回流冷凝管的三口瓶中，加入880g环己烷、20g分散剂SPan-80，升温45℃搅拌30min，得到分散相；将75g海藻酸钠，90g甲基丙烯酸，30％w/w聚乙烯醇水溶液45g，60％w/w戊二醛水溶液24g，分别加入烧杯中常温搅拌10min配制成聚合相，分散相与聚合相之比为3.84：1，把玻璃棒插入三口瓶底部，将聚合相沿着玻璃棒加入分散相，调节搅拌速度，待聚合相分散为均匀珠粒，温度升至58-62℃，向聚合体系加0.2mol/L盐酸水溶液20ml，珠粒定型后将温度升至70-78℃，再加入0.2mol/LNaOH水溶液20ml反应3h，反应完毕过滤出树脂用95％乙醇洗涤至洗脱液清澈，用0.2mol/L碱溶液浸泡8小时，再用蒸馏水洗到中性，用乙醇回流提取24h，真空干燥。
2、测定树脂白球理化指标，方法同实施例2(见表2)
表2实施例2合成的树脂的理化指标
实施例3：
1、合成树脂
在装有搅拌器、温度计、回流冷凝管的三口瓶中，加入850g环己烷、50g分散剂SPan-80，升温45℃搅拌30min，得到分散相；将105g海藻酸钠，150g甲基丙烯酸，30％w/w聚乙烯醇水溶液75g，60％w/w戊二醛水溶液45g，分别加入烧杯中常温搅拌10min配制成聚合相，分散相与聚合相之比为2.14：1，把玻璃棒插入三口瓶底部，将聚合相沿着玻璃棒加入分散相，调节搅拌速度，待聚合相分散为均匀珠粒，温度升至58-62℃，向聚合体系加0.2mol/L盐酸水溶液20ml，珠粒定型后将温度升至70-78℃，再加入0.2mol/LNaOH水溶液20ml反应3h，反应完毕过滤出树脂用95％乙醇洗涤至洗脱液清澈，用0.2mol/L碱溶液浸泡8小时，再用蒸馏水洗到中性，用乙醇回流提取24h，真空干燥。
2、测定树脂白球理化指标，方法同实施例1(见表3)
表3实施例3合成的树脂的理化指标
实施例4：
吸附实验：在20ml具塞三口瓶中，按1ml树脂：2ml血清的比例，加入2ml树脂(实施例1-3)和4ml血清，置于37℃恒温水槽中振摇1小时，吸取血清检测用日本7170型全自动生化分析仪测定吸附前、后血清中脂蛋白含量。
吸附率公式：
式中：cr1—低密度脂蛋白吸附率；c0—吸附前低密度脂蛋白浓度，单位为毫克每升(mg/L)；c1—吸附1h后低密度脂蛋白浓度，单位为毫克每升(mg/L)。
表4实施例1-3的树脂吸附脂蛋白的吸附率
| 树脂 | 脂蛋白吸附率 |
| 实施例1 | 89.75％ |
| 实施例2 | 87.64％ |
| 实施例3 | 88.22％ |
由表1-表4可知，较多的亲水性基团能够增加树脂的吸附率，在三种树脂中较高的羧基能够提高树脂的基本吸附性能，脂蛋白吸附率可达89.75％。
Claims (3)
1.用于吸附血液中低密度脂蛋白的树脂的制备方法，其特征在于，包含以下步骤：
1)配置分散相：在反应容器中加入环己烷和分散剂，温度35-50℃，搅拌25-35min，制得分散相备用；分散剂为SPan-80，分散剂占分散相的质量百分比为2％-5％；
2)配置聚合相：在反应容器中加入海藻酸钠、甲基丙烯酸、聚乙烯醇水溶液和戊二醛水溶液，搅拌至完全溶解，制得聚合相备用；聚合相重量百分比组成为：海藻酸钠25-35％，甲基丙烯酸30-50％，聚乙烯醇水溶液15-25％，戊二醛水溶液8-15％；
3)反向悬浮聚合：将配制好的聚合相缓慢加入分散相，搅拌至聚合相分散为均匀的珠粒，温度升至58-62℃，向聚合体系加0 .1-0 .3mol/L盐酸水溶液，珠粒定型后将温度升至70-78℃，再加入0 .1-0 .3mol/LNaOH水溶液，反应2-4h后，过滤出树脂，用95％乙醇洗涤至洗脱液清澈，用0 .2mol/L碱溶液浸泡6-10h，再用蒸馏水洗至中性，用乙醇回流提取24h，真空干燥；分散相和聚合相的质量比为2-5:1。
2.根据权利要求1所述的用于吸附血液中低密度脂蛋白的树脂的制备方法，其特征在于，所述聚乙烯醇水溶液的质量分数为20-40％，戊二醛水溶液的质量分数为50-70％。
3.权利要求1所述的树脂在吸附低密度脂蛋白中的应用。
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| CN111363166A (zh) * | 2018-12-25 | 2020-07-03 | 天津科技大学 | 一种高强度、自修复抗菌水凝胶及其制备方法 |
| CN113717320A (zh) * | 2020-05-25 | 2021-11-30 | 华熙生物科技股份有限公司 | 富血小板血浆分离凝胶的制备方法及所得产品和应用 |
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| US4781838A (en) * | 1985-08-23 | 1988-11-01 | Commissariat A L'energie Atomique | Solid polyvinyl alcohol-based support able to adsorb lipoproteins and its use for the separation of low density lipoproteins present in a liquid, such as blood plasma |
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| CN111363166A (zh) * | 2018-12-25 | 2020-07-03 | 天津科技大学 | 一种高强度、自修复抗菌水凝胶及其制备方法 |
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