CN109536060A - 一种脱落细胞粘取胶及其制备方法 - Google Patents
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Abstract
本发明公开了一种脱落细胞粘取胶及其制备方法。一种脱落细胞粘取胶，其原料组分包括：松香酯乳液100份，蜡质玉米淀粉5‑8份，糯米粉8‑12份，红薯淀粉1‑2份，木薯淀粉0.5‑0.8份，绿豆淀粉0.3‑0.5份，深薯淀粉0.2‑0.4份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.5‑0.8份，苯酚0.3‑0.5份和聚乙二醇4000 2‑3份，前述份数为质量份数。本发明脱落细胞粘取胶，利用生物可再生资源为原料，制得了一种适于脱落细胞粘取的专用胶，粘度适中，能同时保证脱落细胞粘取率和分离效率，提高了生物分析的准确性；所用原料以生物可再生原料为主，绿色环保、可再生、可降解。
Description
技术领域
本发明涉及一种脱落细胞粘取胶及其制备方法，属于生物领域。
背景技术
目前，在现场脱落细胞提取和实验室检验过程中，采用多种方法，例如剪取法、擦拭法、吸附法以及粘取法来提取生物物证。其中，利用胶带粘附提取技术可以方便地粘取脱落细胞以进行DNA验证或进行试验。
脱落细胞粘取时，如果粘性过大会增大杂质含量，且不易提取分离，进而影响后期分析的准确性，如果粘性过小，会造成脱落细胞采集量不足等问题，进而影响后期分析。而现有技术中并没有专门的脱落细胞粘取胶，且现有的胶需要用到脲醛树脂、酚醛树脂、三聚氰胺甲醛树脂等有毒树脂，同时还存在成本高、不易降解、石化资源消耗大等问题。
发明内容
为了解决现有技术中存在的上述缺陷，本发明提供一种脱落细胞粘取胶及其制备方法。
为解决上述技术问题，本发明所采用的技术方案如下：
一种脱落细胞粘取胶，其原料组分包括：松香酯乳液100份，蜡质玉米淀粉5-8份，糯米粉8-12份，红薯淀粉1-2份，木薯淀粉0.5-0.8份，绿豆淀粉0.3-0.5份，深薯淀粉0.2-0.4份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.5-0.8份，苯酚0.3-0.5份和聚乙二醇4000 2-3份，前述份数为质量份数。
申请人经研究发现，通过不同支链度淀粉的协同促进和改性，可与松香酯乳液混合反应得到粘度适中的脱落细胞粘取专用胶，且所用原料以生物可再生原料为主，绿色环保、可再生、可降解。
为了保证胶的粘结性能，松香酯乳液的固含量为30-40％。
优选，松香酯为松香甲酯、松香乙酯、松香甘油酯、松香季戊四醇酯或马来松香季戊四醇酯中的至少一种。
进一步优选，松香酯为松香季戊四醇酯。这样能得到粘度更加适中的胶，能更好的保证生物分析的准确性。
为了保证松香酯乳液的分散性、稳定性和胶黏性等性能，松香酯乳液所用乳化剂的制备方法包括如下步骤：
1)将松香、氧化镁和聚乙二醇的混合物在温度为250～260℃下反应3-4h，其中，氧化镁的用量为松香质量的0.5-0.8％，聚乙二醇的质量用量为松香质量的0.8-1倍；
2)将步骤1)所得的物料中加入硝酸和对苯二酚，在温度为130-140℃下反应8-12h，即得松香酯乳液所用乳化剂，硝酸的质用量为松香质量的2-3％，对苯二酚的质量用量为松香质量的1.2-1.5％。
乳液制备时，乳化剂的选择是非常关键的，不同物料所用乳化剂不同，本申请以松香为原料在特定条件下制备得到的乳化剂对松香酯具有很好的溶解性和乳化性能，能确保所得乳液的稳定性和分散性，且能得到高固含量的乳液。
为了进一步确保乳化剂的乳化性能，步骤1)中的聚乙二醇为聚乙二醇10000。
为了进一步提高反应效率，步骤2)中的硝酸的质量浓度为20-25％。
为了进一步确保乳液的稳定性，松香酯乳液的制备方法包括如下步骤：
1)将松香酯和乳化剂分别研磨为粒径小于0.5mm的粉末状，其中，乳化剂的质量用量为松香酯质量的10％；
2)将松香酯和乳化剂的混合物，在400-500r/min的搅拌速度下，以10-12℃/min的速度升温熔化，同时以50-60滴/min的速度滴加温度为85-95℃的水；
3)待转相成功后，将温度控制在90-100℃，然后将搅拌速度提升为900-1000r/min，加入十二醇，搅拌3-5min，然后加入温度为85-95℃的水，使乳液的固含量为30-40％，继续保温搅拌10-15min，即得松香酯乳液。
乳液的制备是非常敏感的，任何一反应要素的少许变动，都可能导致制备的失败，尤其是对于软化点比较高的松香酯，乳化难度相对较大，申请人经研究发现，采用上述方法所制备的乳液稳定期大于12个月。
上述脱落细胞粘取胶的制备方法，包括顺序相接的如下步骤:
1)将蜡质玉米淀粉、糯米粉、红薯淀粉、木薯淀粉、绿豆淀粉和深薯淀粉和质量浓度为10％的甲酸水溶液混匀，在温度为60-70℃下搅拌反应2-3h，然后调节pH值为6-7；
2)在温度为60-70℃的搅拌条件下，将山梨酸钾、苯酚和聚乙二醇4000依次加入到步骤1)所得的物料中，加料完毕后继续保温搅拌8-10min；
3)将步骤2)所得的物料升温到80-90℃，搅拌条件下，以100-120滴/min的速度滴入松香酯乳液，滴加完毕后继续搅拌反应3-4h，即得脱落细胞粘取胶。
为了进一步保证所得粘取胶的性能，优选，步骤1)中搅拌速度为100-120r/min；步骤2)中搅拌速度为500-600r/min；步骤3)中搅拌速度为50-60r/min。
使用时，将脱落细胞粘取胶均有涂布在基体上，然后在盖上保护膜，使用时，撕下保护膜即可。
步骤1)中对淀粉混合物改性，使得淀粉表面的活性基团增加，粘结性增大、耐水性提高。
本发明未提及的技术均参照现有技术。
本发明脱落细胞粘取胶，利用生物可再生资源为原料，制得了一种适于脱落细胞粘取的专用胶，粘度适中，能同时保证脱落细胞粘取率和分离效率，提高了生物分析的准确性；所用原料以生物可再生原料为主，绿色环保、可再生、可降解。
具体实施方式
为了更好地理解本发明，下面结合实施例进一步阐明本发明的内容，但本发明的内容不仅仅局限于下面的实施例。
实施例1
一种脱落细胞粘取胶的原料组分包括：松香酯乳液100份，蜡质玉米淀粉5份，糯米粉8份，红薯淀粉1份，木薯淀粉0.5份，绿豆淀粉0.3份，深薯淀粉0.2份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.5份，苯酚0.3份和聚乙二醇4000 2份，前述份数为质量份数。
松香酯乳液的固含量为30-40％，松香酯为松香季戊四醇酯。
松香酯乳液所用乳化剂的制备方法包括如下步骤：
1)将松香、氧化镁和聚乙二醇10000的混合物在温度为250～260℃下反应3.5h，其中，氧化镁的用量为松香质量的0.6％，聚乙二醇10000的质量用量为松香质量的0.9倍；
2)将步骤1)所得的物料中加入硝酸和对苯二酚，在温度为130-140℃下反应10h，即得松香酯乳液所用乳化剂，硝酸的质用量为松香质量的2.5％，硝酸的质量浓度为20％，对苯二酚的质量用量为松香质量的1.3％。
松香酯乳液的制备方法包括如下步骤：
1)将松香季戊四醇酯和上述所得的乳化剂分别研磨为粒径小于0.5mm的粉末状，其中，乳化剂的质量用量为松香酯质量的10％；
2)将松香酯和乳化剂的混合物，在450r/min的搅拌速度下，以11℃/min的速度升温熔化，同时以60滴/min的速度滴加温度为85-95℃的水；
3)待转相成功后(粘度明显减小、有乳状物生成)，将温度控制在90-100℃，然后将搅拌速度提升为1000r/min，加入十二醇，搅拌4min，然后加入温度为85-95℃的水，使乳液的固含量为35％，继续保温搅拌13min，即得松香酯乳液。
脱落细胞粘取胶的制备方法，包括顺序相接的如下步骤:
1)将蜡质玉米淀粉、糯米粉、红薯淀粉、木薯淀粉、绿豆淀粉和深薯淀粉和质量浓度为10％的甲酸水溶液混匀，在温度为60-70℃下搅拌(110r/min)反应2.5h，然后调节pH值为6-7；
2)在温度为60-70℃的搅拌(550r/min)条件下，将山梨酸钾、苯酚和聚乙二醇4000依次加入到步骤1)所得的物料中，加料完毕后继续保温搅拌(60-70℃，550r/min)10min；
3)将步骤2)所得的物料升温到80-90℃，搅拌(55r/min)条件下，以120滴/min的速度滴入松香酯乳液，滴加完毕后继续搅拌反应3.5h，即得脱落细胞粘取胶。
实施例2
与实施例1基本相同，所不同的是：一种脱落细胞粘取胶的原料组分包括：松香酯乳液100份，蜡质玉米淀粉8份，糯米粉12份，红薯淀粉2份，木薯淀粉0.8份，绿豆淀粉0.5份，深薯淀粉0.4份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.8份，苯酚0.5份和聚乙二醇4000 3份，前述份数为质量份数。
实施例3
与实施例1基本相同，所不同的是：一种脱落细胞粘取胶的原料组分包括：松香酯乳液100份，蜡质玉米淀粉6份，糯米粉10份，红薯淀粉1.5份，木薯淀粉0.6份，绿豆淀粉0.4份，深薯淀粉0.3份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.7份，苯酚0.4份和聚乙二醇4000 2.5份，前述份数为质量份数
对比例1
一种脱落细胞粘取胶的原料组分包括：蜡质玉米淀粉6份，糯米粉10份，红薯淀粉1.5份，木薯淀粉0.6份，绿豆淀粉0.4份，深薯淀粉0.3份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.7份，苯酚0.4份和聚乙二醇4000 2.5份，前述份数为质量份数松香酯乳液的固含量为30-40％，松香酯为松香季戊四醇酯。
脱落细胞粘取胶的制备方法，包括顺序相接的如下步骤:
1)将蜡质玉米淀粉、糯米粉、红薯淀粉、木薯淀粉、绿豆淀粉和深薯淀粉和质量浓度为10％的甲酸水溶液混匀，在温度为60-70℃下搅拌(110r/min)反应2.5h，然后调节pH值为6-7；
2)在温度为60-70℃的搅拌(550r/min)条件下，将山梨酸钾、苯酚和聚乙二醇4000依次加入到步骤1)所得的物料中，加料完毕后继续保温搅拌(60-70℃，550r/min)10min；
3)将步骤2)所得的物料升温到80-90℃，搅拌(55r/min)条件下，搅拌反应3.5h，即得脱落细胞粘取胶。
对比例2
与实施例3基本相同，所不同的是：不含蜡质玉米淀粉。
对比例3
与实施例3基本相同，所不同的是：不含糯米粉。
对比例4
与实施例3基本相同，所不同的是：不含深薯淀粉。
应用例
将各例所得的脱落细胞粘取胶均有涂布在基体上，然后在盖上保护膜，其中，所用基体的材质为聚丙烯，保护膜所用材质为聚乙烯薄膜，涂布厚度为25μm，脱落细胞粘取胶的涂布大小为直径为2cm的圆形。
检材：留有唇印的杯子。
粘度为采用粘度仪测量各例所得的脱落细胞粘取胶的粘度；脱落细胞粘取率为所粘取的脱落细胞占杯子上总的脱落细胞的百分比；杂质粘取率为所粘取的物质中的杂质的面积含量；分离效率为将粘取后的基材浸没在TNE缓冲液中，56℃下水浴30min后，基材上脱落细胞的分离效率。
由上表可看出，脱落细胞粘取胶的最佳粘度为10-14Pa·s，更优选为12-13Pa·s，这样同时保证脱落细胞粘取率和分离效率，且通过对比例的结果可看出，原料的选择不仅影响到脱落细胞粘取胶的粘度，而且会影响分离效率。
Claims (10)
1.一种脱落细胞粘取胶，其特征在于：其原料组分包括：松香酯乳液100份，蜡质玉米淀粉5-8份，糯米粉8-12份，红薯淀粉1-2份，木薯淀粉0.5-0.8份，绿豆淀粉0.3-0.5份，深薯淀粉0.2-0.4份，质量浓度为10％的甲酸水溶液100份，山梨酸钾0.5-0.8份，苯酚0.3-0.5份和聚乙二醇4000 2-3份，前述份数为质量份数。
2.如权利要求1所述的脱落细胞粘取胶，其特征在于：松香酯乳液的固含量为30-40％。
3.如权利要求1或2所述的脱落细胞粘取胶，其特征在于：松香酯为松香甲酯、松香乙酯、松香甘油酯、松香季戊四醇酯或马来松香季戊四醇酯中的至少一种。
4.如权利要求3所述的脱落细胞粘取胶，其特征在于：松香酯为松香季戊四醇酯。
5.如权利要求1或2所述的脱落细胞粘取胶，其特征在于：松香酯乳液所用乳化剂的制备方法包括如下步骤：
1)将松香、氧化镁和聚乙二醇的混合物在温度为250～260℃下反应3-4h，其中，氧化镁的用量为松香质量的0.5-0.8％，聚乙二醇的质量用量为松香质量的0.8-1倍；
2)将步骤1)所得的物料中加入硝酸和对苯二酚，在温度为130-140℃下反应8-12h，即得松香酯乳液所用乳化剂，硝酸的质用量为松香质量的2-3％，对苯二酚的质量用量为松香质量的1.2-1.5％。
6.如权利要求5所述的脱落细胞粘取胶，其特征在于：步骤1)中的聚乙二醇为聚乙二醇10000。
7.如权利要求5所述的脱落细胞粘取胶，其特征在于：步骤2)中的硝酸的质量浓度为20-25％。
8.如权利要求5所述的脱落细胞粘取胶，其特征在于：松香酯乳液的制备方法包括如下步骤：
1)将松香酯和乳化剂分别研磨为粒径小于0.5mm的粉末状，其中，乳化剂的质量用量为松香酯质量的10％；
2)将松香酯和乳化剂的混合物，在400-500r/min的搅拌速度下，以10-12℃/min的速度升温熔化，同时以50-60滴/min的速度滴加温度为85-95℃的水；
3)待转相成功后，将温度控制在90-100℃，然后将搅拌速度提升为900-1000r/min，加入十二醇，搅拌3-5min，然后加入温度为85-95℃的水，使乳液的固含量为30-40％，继续保温搅拌10-15min，即得松香酯乳液。
9.权利要求1-8任意一项所述的脱落细胞粘取胶的制备方法，其特征在于：包括顺序相接的如下步骤:
1)将蜡质玉米淀粉、糯米粉、红薯淀粉、木薯淀粉、绿豆淀粉和深薯淀粉和质量浓度为10％的甲酸水溶液混匀，在温度为60-70℃下搅拌反应2-3h，然后调节pH值为6-7；
2)在温度为60-70℃的搅拌条件下，将山梨酸钾、苯酚和聚乙二醇4000依次加入到步骤1)所得的物料中，加料完毕后继续保温搅拌8-10min；
3)将步骤2)所得的物料升温到80-90℃，搅拌条件下，以100-120滴/min的速度滴入松香酯乳液，滴加完毕后继续搅拌反应3-4h，即得脱落细胞粘取胶。
10.如权利要求9所述的制备方法，其特征在于：步骤1)中搅拌速度为100-120r/min；步骤2)中搅拌速度为500-600r/min；步骤3)中搅拌速度为50-60r/min。
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