CN116042216A - 一种黑枸杞碳量子点的制备方法及应用 - Google Patents
一种黑枸杞碳量子点的制备方法及应用 Download PDFInfo
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Abstract
本发明公开了一种黑枸杞碳量子点的制备方法及应用,包括如下步骤:将黑枸杞搅碎并放置于溶剂中进行搅拌,然后放置于反应釜中进行加热,待加热结束冷却至室温,将反应产物进行离心过滤透析,以获得黑枸杞碳量子点。黑枸杞作为一种商业化的商品,成本低廉,易于获取,同时安全性较高,以黑枸杞中富含的花青素作为主要碳源,在水热反应条件下形成量子点,不仅使得量子点成本得到了降低,而且生成的量子点具有较高比表面积,进而具备较高的荧光强度和量子转换效率。此外花青素中富含的氨基酸除了作为量子点形成过程中的辅助碳源,还作为主要氮源,进一步提升了量子产率。
Description
技术领域
本发明涉及一种黑枸杞碳量子点的制备方法及应用,属于碳量子点领域。
背景技术
碳量子点具有良好的光电性能,同时还具有环保低毒等优势,因此受到了广大研究者的关注。但是传统的碳量子点合成,为了保证最终产物的量子转化率,都是使用纯度较高的一种或者多种原料,致使碳量子点的成本难以进一步下降,如果采用纯度较低的原料进行合成,又会导致量子转化率的下降,因此如何获得一种成本低廉同时量子转化率较高的碳量子点是现阶段一个十分重要的课题。
发明内容
本发明所要解决的技术问题在于克服现有技术的不足而提供一种黑枸杞碳量子点的制备方法及应用。
解决上述技术问题,本发明采用如下技术方案:、
一种黑枸杞碳量子点的制备方法,包括如下步骤:将黑枸杞搅碎并放置于溶剂中进行搅拌,然后放置于反应釜中进行加热,待加热结束冷却至室温,将反应产物进行离心过滤透析,以获得黑枸杞碳量子点。
本发明的有益效果为:
黑枸杞作为一种商业化的商品,成本低廉,易于获取,同时安全性较高,以黑枸杞中富含的花青素作为主要碳源,在水热反应条件下形成量子点,不仅使得量子点成本得到了降低,而且生成的量子点具有较高比表面积,进而具备较高的荧光强度和量子转换效率。此外花青素中富含的氨基酸除了作为量子点形成过程中的辅助碳源,还作为主要氮源,进一步提升了量子产率。
本发明所述溶剂为去离子水。
发明加热温度为160-180℃。
本发明加热时间为24h。
本发明所述黑枸杞产地为青海。
一种黑枸杞碳量子点的应用,将黑枸杞碳量子点的制备方法制备得到的黑枸杞碳量子点溶解稀释,并与待测溶液混合,通过对待测溶液的荧光图像进行分析,以对溶液中是否存在Ag+进行检测。
本发明通过对待测溶液的荧光强度进行测量,从而获得Ag+的浓度。
一种黑枸杞碳量子点的应用,将黑枸杞碳量子点的制备方法制备得到黑枸杞碳量子点溶解稀释,而后倒入培养皿中参与成纤维细胞配合,通过分析培养皿的荧光图像,从而对成纤维细胞进行定位。
本发明的其他特点和优点将会在下面的具体实施方式、附图中详细的揭露。
附图说明
下面结合附图对本发明做进一步的说明:
图1为本发明实施例1的黑枸杞碳量子点的TEM图;
图2为图1提高分辨率后的TEM图;
图3为本发明实施例1的黑枸杞碳量子点的粒径尺寸统计分布图;
图4为本发明实施例1的黑枸杞碳量子点在S iO2/S i衬底上的AFM图;
图5为本发明实施例1的黑枸杞碳量子点的高度统计分布图;
图6为本发明实施例1的黑枸杞碳量子点的XPS图;
图7为本发明实施例1的黑枸杞碳量子点的高分辨率XPS图(N 1s);
图8为本发明实施例1的黑枸杞碳量子点的在不同pH条件下的归一化荧光强度;
图9为本发明实施例1的黑枸杞碳量子点的在经过不同时间后的归一化荧光强度;
图10为本发明实施例1的黑枸杞碳量子点的在不同浓度NaC l溶液中的归一化荧光强度;
图11为本发明实施例1的黑枸杞碳量子点在紫外灯照射下荧光强度随时间变化;
图12为本发明实施例1的黑枸杞碳量子点配置不同浓度水溶液对成纤维细胞进行培养后的成纤维细胞存活率;
图13为本发明实施例1的成纤维细胞经过黑枸杞碳量子点水溶液培养后在共焦显微镜下的图像;
图14为本发明实施例1的黑枸杞碳量子点在不同溶液中荧光强度比对图;
图15为本发明实施例1黑枸杞碳量子点的溶液在不同pH条件下添加银离子前后荧光强度相对变化量的对应关系图;
图16为本发明实施例1黑枸杞碳量子点的溶液(具有不同银离子浓度)在365nm紫外光照条件下照片;
图17为本发明实施例1黑枸杞碳量子点的溶液(具有不同银离子浓度)在365nm紫外光照条件下的RGB数值变化图;
图18为本发明采用不同溶剂制备的黑枸杞碳量子点溶于水后对成纤维细胞进行培养后,成纤维细胞存活率数据图(浓度均为300μg/ml);
图19为本发明采用不同产地的黑枸杞制备的黑枸杞碳量子点的量子效率图。
具体实施方式
下面结合本发明实施例的附图对本发明实施例的技术方案进行解释和说明,但下述实施例仅为本发明的优选实施例,并非全部。基于实施方式中的实施例,本领域技术人员在没有做出创造性劳动的前提下所获得其他实施例,都属于本发明的保护范围。
在下文描述中,出现诸如术语“内”、“外”、“上”、“下”、“左”、“右”等指示方位或者位置关系仅是为了方便描述实施例和简化描述,而不是指示或暗示所指的装置或者元件必须具有特定的方位、以特定的方位构造和操作,因此不能理解为对本发明的限制。
实施例1:
本实施例提供了一种黑枸杞碳量子点的制备方法,包括如下步骤:将3.0g黑枸杞搅碎并倒入20.0ml去离子水中搅拌30mi n,然后转移至50.0ml反应釜中进行加热,加热时间为180℃,加热时间为24h,加热结束后待反应釜冷却至室温,将获得的溶液在8000rpm条件下离心15mi n,将离心得到的固体用0.22μm孔径的过滤膜进行过滤,以去大颗粒固体,而后用透析袋(MWCO:100-500Da)透析48h,以获得黑枸杞碳量子点。
得到的黑枸杞碳量子点于4℃条件下长期保存。
本实施例中黑枸杞的原料产地为青海柴达木,品牌为福茗源,产品标准号为GH/T1091。
参见图1-2,可以看到黑枸杞碳量子点形成了蜂窝结构,说明黑枸杞碳量子点产生了高度石墨化,具备较大的比表面积。参见图3,本实施例黑枸杞碳量子点的尺寸范围在3.0-6.5nm之间,平均值在4.5nm。参见图4-5,黑枸杞碳量子点的平均高度为0.73nm。黑枸杞碳量子点的小尺寸配合蜂窝结构,极大提升了比表面积,提升了反应活性。
参见图6,在285、400、533eV三个峰位分别对应C、N、O三种元素,同时通过三种元素含量可以判断出,C、N来源于黑枸杞,而O不仅来自于黑枸杞,还来自于去离子水,说明水热反应过程中去离子水不仅作为溶剂,还作为了反应物之一。参见图7,可以看到在黑枸杞碳量子点中的N来源于吡啶酸(399.8eV)、氨基(400.2V)以及吡咯(402.1eV),说明黑枸杞中的氨基酸(主要为天冬氨酸、谷氨酸、丙氨酸和脯氨酸)在黑枸杞碳量子点的形成过程中具有十分重要的作用。由此可见,黑枸杞中的花青素以及氨基酸最终在黑枸杞碳量子点的表面形成了大量含氮和含氧的功能性基团。
参见图8,黑枸杞碳量子点溶解在酸性溶液和碱性溶液中的荧光强度相较溶解在中性溶液中时均较低,说明酸碱环境下使得黑枸杞碳量子点表面功能性基团产生了质子化或者反质子化。
参见图9,黑枸杞碳量子点溶解在去离子水中后,荧光强度随着经过时间的增加仅发生了略微衰减,在第8天衰减了10%左右,参见图10,黑枸杞碳量子点溶解在氯化钠溶液中时,荧光强度几乎不受到氯化钠浓度影响,参见图11,黑枸杞碳量子点溶解在去离子水中后在紫外照射16h,荧光强度同样几乎没有变化。这可能是黑枸杞碳量子点表面的功能性基团在中性环境下具备较强的稳定性,有效提升了黑枸杞碳量子点的应用场景。
本实施例的黑枸杞碳量子点量子产率高达21.8%,远高于现有的其他碳量子点量子产率。
本实施例的黑枸杞碳量子点能够用于细胞标记。
具体的,将成纤维细胞放置在18×18mm2的载玻片上,而后放入培养皿中,然后在培养皿中配置质量分数为10%的胎牛血清、100mg/ml的谷氨酰胺、100mg/ml的丙酮酸钠、青霉素(100un its/mL)、链霉素(100un its/mL)、黑枸杞碳量子点的水溶液,在37℃空气加湿条件下对成纤维细胞进行培养24h。而后使用Ze i ss LSM 780共焦激光扫描荧光显微镜对载玻片进行拍摄。参见图12,可以发现,随着成纤维细胞培养过程中使用的黑枸杞碳量子点的水溶液浓度增加,细胞成活率仅有非常细微的下降,即使黑枸杞碳量子点的水溶液浓度达到300μg/ml,成纤维细胞依然有95%左右的成活率,由此可见,黑枸杞碳量子点具有良好的细胞兼容性,几乎没有任何细胞毒素。参见图13,可以明确发现细胞发出绿色荧光,说明黑枸杞碳量子点被成纤维细胞吸收,且未被代谢分解,由此可见,黑枸杞碳量子点可有效作为一种细胞追踪定位标记物质使用。
参见图18,在其他实施例中,还分别制备了其他不同的黑枸杞碳量子点,区别仅在于黑枸杞是与不同的溶剂混合形成黑枸杞碳量子点的原料。可以看到,在采用乙醇和黑枸杞制备黑枸杞碳量子点对纤维细胞培养,尚且能满足84%左右的成活率,但是使用甲酰胺、乙酸乙酯分别制备出的黑枸杞碳量子点对纤维细胞培养,成活率都只有70%左右,说明甲酰胺、乙酸乙酯制备的黑枸杞碳量子点已经对纤维细胞产生了较为明显的毒性。尤其是二甲基亚砜和甲苯制备的黑枸杞碳量子点,细胞成活率都仅有50%和60%,说明二者对制备的黑枸杞碳量子点对细胞已经表现出较为剧烈的毒性了。
本实施例的黑枸杞碳量子点还能够用于Ag+检测。
配置多份溶液,每份溶液包含Al3+,Ca2+,Cd2+,Cr3+,Co2+,Cu2+,Fe2+,Hg2+,K+,Mg2+,Mn2 +,Na+,N i2+,Pb2+和Zn2+中的一种离子,且各个溶液中上述离子浓度均为120μM,同时还准备了去离子水,分别测试各个溶液和离子水的荧光强度,具体强度参见图14中颜色较深的柱,可以看到各个溶液和去离子水的荧光强度相差无几,说明上述离子均不能有效影响荧光强度。而后将银离子分别加入至各个溶液和离子水中再次测试荧光强度(各个溶液和去离子水中银离子浓度也为120μM),具体强度参见图14中颜色较浅的柱,可以看到所有溶液和离子水的荧光强度全都明显下降,且下降后的荧光强度均相差无几,说明银离子有效降低荧光强度,故而黑枸杞碳量子点针对银离子检测具有极好的特异性。
参见图15,向不同pH值的溶液中添加银离子,F0表示添加银离子前的荧光强度,F表示添加银离子后的荧光强度,可以发现,当pH=7时F0和F的差距最大,说明黑枸杞碳量子点最适宜在中性溶液中对银离子进行检测。
在此基础上,在一未知溶液中添加黑枸杞碳量子点,而后添加银离子,通过观测银离子添加前后的荧光强度相对变化量,可以对未知溶液的pH值酸碱度进行判断,相对变化量越大,则说明溶液越接近中性,相对变化量越小,则说明溶液酸性或者碱性越强。
参见图16,在紫外照射下,黑枸杞碳量子点发出绿色荧光,但是随着银离子浓度增加,荧光强度逐渐下降。参见图17,可以发现,银离子浓度在0-20μM范围内,银离子浓度的RGB数值呈近乎线性关系,故而黑枸杞碳量子点在对未知溶液进行银离子检测的时候,可以根据RGB数值预估Ag+的浓度。此外还可以发现,黑枸杞碳量子点对银离子的浓度检测下限为59nM。
由于黑枸杞中成分多样,且产地不同也会导致黑枸杞的具体成分和各成分含量比例发生变化,为了确认是否所有产地的黑枸杞制备的黑枸杞碳量子点都具有良好的荧光效率,还分别额外使用宁夏红枸杞、中华红枸杞、宁夏黑枸杞、甘肃黑枸杞、新疆黑枸杞分别制备的量子点与本实施例的青海黑枸杞制备的量子点进行比较。其中新疆黑枸杞产地为新疆精河,品牌为欣果果。宁夏黑枸杞产地为宁夏银川,品牌为杞里香。
参见图19,宁夏红枸杞和中华红枸杞制备的量子点的量子产率仅为10-12%左右,与现有常规的碳量子点量子产率持平,并无明显优势。但是宁夏黑枸杞、甘肃黑枸杞、新疆黑枸杞制备的量子点量子产率在15-18%左右,特别的,青海黑枸杞制备的量子点量子产率甚至高达22%,几乎是红枸杞制备的量子点的两倍,同时也明显高于其他产地黑枸杞制备的量子点。由此可见,为了相较现有技术而言获得更高量子产率的量子点,其原料仅可以选用黑枸杞,尤其是青海黑枸杞,和其他所有枸杞之间在量子产率上都拉开了明显差距,说明不同产地的枸杞对最终量子产率具有不可忽视的影响。除了量子产率之外,还可以注意到采用不同产地的枸杞制备出的量子点,其荧光强度峰值所对应的波长也存在变化,黑枸杞制备量子的荧光峰位波长全都高于红枸杞制备量子的荧光峰位波长,说明黑枸杞制备量子点的荧光峰位波长存在明显的红移,这就使得其荧光对生物组织损伤更小,穿透深度达,也就使得黑枸杞制备的量子点具备了更加广阔的应用前景。
以上所述,仅为本发明的具体实施方式,但本发明的保护范围并不局限于此,熟悉该本领域的技术人员应该明白本发明包括但不限于附图和上面具体实施方式中描述的内容。任何不偏离本发明的功能和结构原理的修改都将包括在权利要求书的范围中。
Claims (8)
1.一种黑枸杞碳量子点的制备方法,其特征在于,包括如下步骤:将黑枸杞搅碎并放置于溶剂中进行搅拌,然后放置于反应釜中进行加热,待加热结束冷却至室温,将反应产物进行离心过滤透析,以获得黑枸杞碳量子点。
2.根据权利要求1所述的黑枸杞碳量子点的制备方法,其特征在于,所述溶剂为去离子水。
3.根据权利要求2所述的黑枸杞碳量子点的制备方法,其特征在于,加热温度为160-180℃。
4.根据权利要求2所述的黑枸杞碳量子点的制备方法,其特征在于,加热时间为24h。
5.根据权利要求2所述的黑枸杞碳量子点的制备方法,其特征在于,所述黑枸杞产地为青海。
6.一种黑枸杞碳量子点的应用,其特征在于,将权利要求1或2或3或4所述的黑枸杞碳量子点的制备方法制备得到的黑枸杞碳量子点溶解稀释,并与待测溶液混合,通过对待测溶液的荧光图像进行分析,以对溶液中是否存在Ag+进行检测。
7.根据权利要求4所述的黑枸杞碳量子点的应用,其特征在于,通过对待测溶液的荧光强度进行测量,从而获得Ag+的浓度。
8.一种黑枸杞碳量子点的应用,其特征在于,将权利要求1或2或3所述的黑枸杞碳量子点的制备方法制备得到黑枸杞碳量子点溶解稀释,而后倒入培养皿中参与成纤维细胞配合,通过分析培养皿的荧光图像,从而对成纤维细胞进行定位。
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