CN110684014B - 可用于卵巢癌的一种具有聚集诱导发射效应的水溶性荧光探针和纳米粒及其制备方法和应用 - Google Patents
可用于卵巢癌的一种具有聚集诱导发射效应的水溶性荧光探针和纳米粒及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种具有聚集诱导发射（AIE）效应的水溶性荧光探针和纳米粒及制备方法和应用，该荧光探针分子含有两个甲氧苯基和一个苯基取代的咪唑环及两个季铵盐结构特征，在水中具有良好的分散性，能够形成胶束，并且具有聚集诱导发射效应，斯托克斯位移大等特点，适应于荧光检测；在溶血磷脂酸的诱导下，该探针能够自组装，从而聚集产生黄色荧光，通过测定发射光的强度可实现溶血磷脂酸的定量分析，具有优良的特异性，高灵敏度，抗干扰性强等优点，特别适用于化学溶液体系，生物样品等环境中的溶血磷脂酸的检测，对于痕量溶血磷脂酸的检测和卵巢癌患者血浆中溶血磷脂酸的含量的测定及临床早期诊断和用药指导具有十分重要的实用价值。
Description
技术领域
本发明涉及荧光探针及纳米粒，特别涉及一种具有聚集诱导发射效应的水溶性的荧光分子探针或荧光纳米粒，及它们的制备方法，特别还涉及该荧光分子探针纳米粒在溶血磷脂酸的检测方面的应用，属于化学分析、生物分析检测技术领域。
背景技术
溶血磷脂酸（lysobisphosphatidic acids，LPA）是迄今发现的最小、最简单的甘油磷脂，由含磷酸的亲水端和含长脂肪链的疏水端组成。它是血清的正常成分，在血小板聚集过程中由血小板产生。溶血磷脂酸作为一种细胞膜脂类衍生物和代谢中间体，介导着广泛的生化，生理和病理过程，如刺激细胞增殖、分化和迁移，平滑肌收缩，神经递质释放和促进血小板的聚集。最近有研究表明，卵巢癌细胞产生LPA，LPA本身也可作为卵巢癌活化因子。在血浆中LPA的正常生理浓度约为0.1 µM~6.3 µM，而在卵巢癌患者中血浆LPA水平显著升高，因此升高的血浆LPA水平可以用作检测人类卵巢癌的潜在生物标志物。LPA的临界值为1.3 µM，其对卵巢癌患者诊断的敏感性和特异性分别高达95%和92%。（X. Fang, D.Gaudette, T. Furui, M. Mao, V. Estrella, A. Eder, T. Pustilnik, T. Sasagawa,R. Lapushin, S. Yu, R.B. Jaffe, J.R. Wiener, J.R. Erickson and G.B. Mills,Ann. N.Y. Acad. Sci., 2000, 905, 188; Y. Xu, Z. Shen, D. W. Wiper, M. Wu,R.E. Morton, P. Elson, A.W. Kennedy, J. Belinson, M. Markman and G. Casey, J.A. M. A. 1998, 280, 719.）。卵巢癌是一种高度转移性疾病，以腹膜广泛传播和腹水为特征，是妇科恶性肿瘤死亡的主要原因，对女性生命造成严重威胁。由于卵巢癌早期症状不明显，大约 60%的卵巢癌在晚期才被诊断出来，治疗效果受到影响。因此在卵巢癌早期如果可以通过筛查得到检测并且及时进行治疗，就可以在对抗该顽疾时占上风，实现有效的卵巢癌的早期诊断对于提高总体生存至关重要（Anonymous, Lancet, 2016, 387, 918.）。由此可见，开发简单有效的特异性检测LPA的方法对于卵巢癌的预防和诊断具有迫切性和重要的意义。
迄今为止已经开发了一些用于检测血浆LPA水平的方法，经典方法包括电喷雾质谱（ESI-MS），基质辅助激光解吸电离质谱（MALDI-MS），GC-MS等，尽管分析灵敏度很高，但这些方法通常需要费力的样品预处理，样品衍生化或长时间的分离，并且需要昂贵的仪器来完成，不适用于常规诊断（Y.J. Xiao, Y.H. Chen, W. K. Alexander, B. Jerome and Y.Xu, Ann. Ny. Acad. Sc., 2000, 905, 242; T. Tanaka, H. Tsutsui, K. Hirano, T.Koike, A. Tokumuraand K. Satouchi, J. Lipd. Res., 2004, 45, 2145.）。近年来一些新型的定量分析LPA的方法也被开发，例如毛细管电泳间接紫外检测法，酶联免疫法（Elisa）。虽然这些方法具有特异性，仪器相对简单，但它们操作步骤较多。市面上已经用于LPA检测的Elisa试剂盒，由于其所用到的酶试剂容易受到环境影响，且有的底物易挥发或者对光敏感，操作条件比较严格。并且有些操作步骤需要进行温育30分钟或者1小时，相对比较繁琐，具有一定的局限性。荧光探针由于其高灵敏度，简单方便和实时分析的特点引起了广泛关注，但LPA化学传感器的例子很少见。Wu Junchen等人报道了一种可以特异性识别LPA的多肽探针，它可以在体内对LPA特异性结合浓聚在肿瘤部位，也能够在体外对血清中的LPA识别发出增强的荧光，但该探针的研究显示是在LPA浓度为6~20 µM范围内有良好的线性关系，如果将其作为一种实时快速的即时检验法测定LPA用于卵巢癌的临床诊断，LPA浓度超出该范围就无法得到精确的检测结果（吴君臣.对溶血磷脂酸特异性识别的多肽探针及其制备与应用：中国, 106518965 A[P]. 2017. 03. 22.）。LanMinhuan等人设计了一种基于3-苯基噻吩的水溶性共聚噻吩（CPT9）用于荧光检测溶血磷脂酸，它是利用LPA具有丰富的负电荷和较长疏水链的特点来设计的基于静电作用，疏水作用和氢键的特异性识别LPA的荧光探针。虽然具有高灵敏度和高选择性，但这一探针是一种共轭聚合物，合成步骤较多，相比于小分子荧光探针，背景噪音可能会较高。（M.H. Lan, W.M. Liu, Y. Wang,J.C. Ge, J.C. Wu, H.Y. Zhang, J.H. Chen, W.J. Zhang and P.F. Wang, ACS Appl.Mater. Interfaces, 2013, 5, 2283.）。另外，这些少数可以用于检测LPA的荧光探针在缓冲液中易形成聚集体而产生聚集导致荧光淬灭效应（aggregation-caused quenching,ACQ），这样就会存在时间和空间上的不稳定性。所以，设计水溶性荧光探针能够高灵敏度和高选择性地方便实时定量分析LPA，并且可以克服聚集导致荧光淬灭效应这一缺陷十分关键。
发明内容
为了克服现有检测溶血磷脂酸（LPA）的荧光探针的性能存在的不足，本发明的第一个目的是在于提供一种对卵巢癌标志物溶血磷脂酸特异性识别的水溶性好，且同时具有聚集诱导发射效应（AIE）的荧光分子探针。
本发明的第二个目的在于提供一种对LPA特异性识别的具有聚集诱导发射效应的能在水溶液中自组装形成纳米粒的荧光分子探针。
本发明的第三个目的在于提供一种原料易得、操作简便的制备所述荧光分子探针的方法。
本发明的第四个目的在于提供一种操作简便、条件温和的制备荧光探针纳米粒的方法。
本发明的第五个目的在于提供一种所述的荧光探针在检测血液中LPA的应用；所述的荧光分子探针在水溶液中自组装形成粒子大而结构疏松的纳米粒，通过LPA与其静电相互作用和疏水作用力，使荧光探针聚集而限制了探针分子的旋转而发射荧光从而实现对LPA的检测，表现出高灵敏度、高选择性和稳定性好的特点。
为了实现上述技术目的，本发明提供了一种对LPA特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针，具有式1结构:
式1
本发明所述的对LPA特异性识别的荧光分子探针具有聚集诱导发射效应（AIE）特性；在400 nm激发下，在500-600 nm处发出强烈的荧光，具有较大的斯托克斯位移。
本发明还提供了一种对LPA特异性识别的荧光分子探针纳米粒，是由所述荧光分子探针自组装形成。
本发明还提供了一种制备所述的对LPA特异性识别的的方法，它包括如下步骤：
1）4-甲酰基吡啶、苯胺和茴香偶酰在醋酸/醋酸铵体系中进行一锅法环化反应，得到式2中间体；
2）式2中间体与1,12-二溴十二烷进行亲核取代反应，得到式3中间体；
3）式3中间体与吡啶发生亲核取代反应即得目标产物；
式2
式3
具体的制备步骤：1）中的反应过程为：将4-甲酰基吡啶和苯胺类化合物溶解在冰醋酸溶剂中于室温下搅拌0.5~1.5小时，再加入苯偶酰类化合物和醋酸铵，在120℃条件下反应6~12小时；
2）中的反应条件为：使用乙腈作为溶剂，在90℃条件下反应6~10小时；
3）中的反应条件为：使用吡啶作为溶剂，在90℃条件下反应6~10小时。
本发明还一种制备所述的对LPA特异性识别的有机荧光分子探针纳米粒的方法，将所述的荧光分子探针溶于有机溶剂后，加入到水溶液中，进行超声处理，即得荧光分子探针纳米粒。
优选的方案，有机溶剂选自甲醇、乙醇、二甲基甲酰胺、四氢呋喃、二甲亚砜、丙酮、乙腈中的至少一种。
所述的水溶液选自纯水、生理盐水、磷酸缓冲液（PBS）、三（羟甲基）氨基甲烷盐酸盐缓冲溶液或4-羟乙基哌嗪乙磺酸缓冲溶液。
本发明还提供了所述对LPA特异性识别的荧光纳米探针的应用，对LPA的检测用于诊断卵巢癌。
优选的方案，将有机荧光纳米探针应用于化学溶液体系、受检者血液中LPA的荧光定量分析检测。
本发明的荧光探针具有四取代咪唑环及四级铵盐结构，中间通过较长的疏水性脂肪链连接。荧光探针分子具有良好的水溶性，在水溶液中可以自组装形成结构疏松的纳米粒，并且不显示荧光。与LPA识别后，由于LPA具有高负电荷和长的疏水脂肪链，可以通过分子间静电作用和疏水作用诱导荧光探针发生聚集导致荧光分子旋转受限而产生强的黄色荧光。其检测原理如图5。
本发明的对LPA特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针的制备路线如下：
本发明的特异性识别LPA的具有聚集诱导发射效应的水溶性荧光分子探针的具体制备方法通过如下步骤实现：
a、将4-甲酰基吡啶和苯胺溶于冰醋酸中，于常温下搅拌反应1小时。然后依次加入茴香偶酰和醋酸铵混合均匀，继续加热至120℃回流反应过夜。TLC检测反应完全后用水淬灭反应，用氢氧化钠溶液调pH至近中性，观察到有絮状沉淀产生，减压过滤得到滤饼，干燥，柱色谱分离纯化得到化合物1；
b、将化合物1溶于乙腈中，加入1,12-二溴十二烷，将体系混合均匀，于90℃加热回流，反应结束后，减压蒸馏除掉溶剂，柱色谱分离纯化得到化合物2；
c、将化合物2溶于吡啶中，体系加热至90℃反应，反应结束后减压蒸馏除掉溶剂，柱色谱分离纯化得到目标化合物。
本发明的对溶血磷脂酸特异性识别的荧光分子探针纳米粒的制备方法如下步骤实现：
将目标化合物充分溶解于有机溶剂中制备成母液，然后用移液枪吸取母液在超声条件下加入到一定量的水溶液中，体系常温搅拌30min后即得检测溶血磷脂酸用的有机纳米粒。利用动态光散射（DLS）和扫描电镜（SEM）来观察形成纳米粒的粒径和形貌。
本发明的技术方案基于荧光分子探针纳米粒检测溶血磷脂酸是充分利用荧光分子探针在水溶液中自组装形成结构疏松的纳米粒，且形成的纳米粒溶液没有荧光，在溶血磷脂酸存在时，通过与溶血磷脂酸的静电相互作用和疏水作用力，使荧光探针聚集形成聚集态而限制了探针分子的旋转导致发射荧光。本发明
正是基于此而建立检测溶血磷脂酸的方法，该方法具有高选择性，高灵敏度，可以广泛推广应用。
本发明的荧光分子探针纳米粒能够用于化学模拟生物体系中溶血磷脂酸的检测；也可用于临床医学上血液中溶血磷脂酸的检测。
与现有技术相比，本发明的有益技术效果是：
1）本发明的对溶血磷脂酸特异性识别的荧光分子探针及纳米粒具有水溶性好，可以自组装形成无荧光的纳米粒的特点，且具有聚集诱导发射效应。与溶血磷脂酸作用时能够形成聚集态，产生557 nm的荧光，特别适用于荧光检测。
2）本发明的对溶血磷脂酸特异性识别的荧光分子探针及纳米粒的制备工艺简单，成本低，有利于大规模生产。
3）本发明的荧光分子探针纳米粒用于检测血液中的溶血磷脂酸，该探针对溶血磷脂酸具有很好的选择性，血清白蛋白，脯氨酸，甘油，磷脂酸，溶血磷脂酰胆碱等对溶血磷脂酸的检测没有干扰，该纳米探针溶液的荧光强度与溶血磷脂酸的浓度在一定的范围内（溶血磷脂酸的浓度范围0-33 µM）具有良好的线性关系，展现出定量检测特性，可以满足临床上对患者血液中溶血磷脂酸的含量检测的要求。
4）本发明的荧光分子探针纳米粒对溶血磷脂酸的响应时间短，测定的灵敏度高，选择性强，具有良好的稳定性，并且操作简单高效，成本低，使得其适合推广运用。
附图说明
【图1】本发明中制备的荧光分子探针纳米粒的动态散射图；
【图2】本发明中制备的荧光分子探针纳米粒与溶血磷脂酸识别的扫描电镜图；
【图3】本发明中荧光分子探针纳米粒的荧光强度与溶血磷脂酸浓度的线性关系，横坐标为溶血磷脂酸浓度，纵坐标为荧光强度；
【图4】本发明中荧光分子探针纳米粒对溶血磷脂酸的选择性；
【图5】为荧光探针纳米粒用于溶血磷脂酸检测的原理图。
具体实施方式
以下实施方式旨在进一步阐释说明发明。
实施例1
本发明的聚集诱导荧光发射增强的化合物3的合成，合成路线如下：
化合物1的合成：分别称取4-甲酰基吡啶（107.1 mg，1 mmol）和苯胺（93.1 mg，1mmol）溶于6~8 mL冰醋酸中，于常温下搅拌1小时。茴香偶酰（207.2 mg，1 mmol）和醋酸铵（462.5 mg，6 mmol）依次加入到反应体系中，在120℃条件下反应过夜，用水淬灭反应，将反应体系倒入200 mL冰水中，用0.1 mmol/L的氢氧化钠溶液将体系pH调至中性，混合物过滤用水洗三次，真空干燥后硅胶柱层析分离纯化得到产物。收率为21.9%。核磁共振分析结果为：1H NMR (500 MHz, CDCl3) δ 8.47 (d,2H), 7.55 (d,2H), 7.45–7.25 (m, 5H), 7.09(t, 2H), 7.05 (d,2H), 6.83 (d,2H), 6.78 (d, 2H), 3.79 (d, 6H).13C NMR（100MHz,CDCl3）δ 159.40, 158.64, 149.59, 143.36, 136.80, 132.29, 126.77, 122.18,113.95, 113.71, 55.20,55.13.
化合物2的合成：称取化合物1（191.5 mg，0.44 mmol）和1,12-二溴十二烷（145.0mg，0.44 mmol）溶解在3 mL乙腈溶液中，室温充分搅拌，反应体系在90℃充分回流8小时，TLC板监测反应完全后，减压蒸馏除去溶剂，硅胶柱层析分离得到化合物2。收率为34.7%。核磁共振分析结果为：1H NMR(500 MHz, DMSO-d6) δ 8.88 (d, 2H), 7.74 (d, 2H), 7.49(dd, 7H), 7.20 (d, 2H), 6.90 (d, 4H), 4.45 (t, 2H), 3.73 (d, 6H), 3.51 (t,2H), 1.79 (dd,4H), 1.23 (s, 16H).13C NMR（125MHz, DMSO-d6）δ 159.99, 159.14,132.75, 130.42, 130.10, 128.97, 128.24, 126.12, 123.70, 121.22, 114.57,114.37, 60.38, 55.57, 35.67, 32.68, 30.77, 29.31, 29.28, 29.18, 28.78, 28.55,27.95, 25.82.
化合物3的合成：称取化合物2（155.0 mg，0.20 mmol）溶解在4~5 mL吡啶溶液中，室温充分搅拌，反应体系在90℃充分回流8小时，TLC板监测反应完全后，减压蒸馏除去溶剂，硅胶柱层析分离得到目标产物3，收率为68.2%。核磁共振分析结果为：1H NMR (500MHz, CD3OD) δ 9.11 – 8.98 (m, 2H), 8.85 – 8.66 (m, 2H), 8.60 (t,1H), 8.12 (s,2H), 7.86 (d, 2H), 7.50 (d, 5H), 7.38 (s, 2H), 7.13 (d, 2H), 6.84 (s, 4H),4.66 (s, 2H), 4.50 (s, 2H), 3.89 – 3.62 (m, 6H), 1.99 (d, 4H), 1.56 – 1.04(m, 16H).13C NMR（125MHz,CD3OD）δ 160.33, 159.48, 145.45, 144.54, 144.41,143.81, 143.77, 141.37, 139.62, 139.58, 136.00, 132.17, 132.14, 129.91,129.88, 128.32, 128.10, 123.66, 120.69, 113.79, 113.76, 113.41, 113.39,61.72, 60.69, 54.38, 54.33, 31.10, 30.84, 30.81, 29.76, 29.72, 29.36, 29.33,29.13, 29.05, 28.68, 25.77.
实施例2
水溶性有机荧光分子探针纳米粒的制备：
用移液枪取40 µL化合物3（1 mM）的磷酸缓冲溶液于ep管中，超声条件下再加入1960 µL 的磷酸缓冲溶液（pH=7.4）使化合物3的最终浓度为20 µM。室温条件下搅拌30 min体系有乳光产生，为了验证其纳米聚集行为，动态光散射实验测定其平均粒径为600 nm，如附图1所示。
实施例3
制备制备的荧光分子探针纳米粒与溶血磷脂酸识别的扫描电子显微镜（SEM）测试：
用移液枪取40 µL化合物3（1 mM）的磷酸缓冲溶液于ep管中，超声条件下再加入1900 µL 的磷酸缓冲溶液（pH=7.4），再加入60 µL溶血磷脂酸的磷酸缓冲液（1 mM）使溶液中化合物3的最终浓度为20 µM，溶血磷脂酸的最终浓度为30 µM，室温条件下搅拌30min。吸取一滴所制备的溶液，滴于铜网上，用滤纸吸干水分，自然晾干，置于投射电镜中进行观察。投射电镜图片如附图2所示。
实施例4
荧光分子探针纳米粒的荧光强度与溶血磷脂酸浓度的线性关系：
向实施例2中所制备的体系中分别加入一系列不同体积的溶血磷脂酸的磷酸缓冲溶液母液，使溶血磷脂酸的最终浓度分别达到，0 µM、0.2 µM、0.4 µM、0.8 µM、1.0 µM、1.2µM、1.4 µM、1.8 µM、2.0 µM、4.0 µM、8.0 µM、10.0 µM、12.0 µM、14.0 µM、16.0 µM、18.0 µM、20.0 µM、25.0 µM、30.0 µM、33.0 µM。所有测试溶液配制完成后，利用涡旋仪使其混合均匀。室温孵育1 min后测量其在557处的荧光发射强度。所得结果如附图3所示，体系中557nm的荧光强度与溶血磷脂酸的浓度在0-33 µM之间有很好的线性关系。
实施例5
有机荧光纳米探针对溶血磷脂酸检测的选择性：
使用制备的荧光分子探针溶液，并评价该探针对溶血磷脂酸的选择性，该纳米探针的激发波长为400 nm。该体系中化合物3的浓度为10 µM，分别向该体系中加入溶血磷脂酸使其最终浓度为50 µM，以及最终浓度为1 mM的L-脯氨酸，甘油，磷酸钠，硫酸镁，乙酸钠，硝酸钠，氟化钠，葡萄糖，尿素，溶血磷脂酰胆碱（LPC）混合充分后，室温孵育1 min后，测其荧光发射光谱并记录557 nm处荧光发射的强度。结果如图4所示，只有加入溶血磷脂酸的体系中产生强的黄色荧光，而加入其它物种的溶液中几乎观察不到黄色荧光，上述结果表明，该有机荧光分子探针纳米粒检测溶血磷脂酸具有良好的选择性和实际应用性。
Claims (6)
1.一种对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针，其特征在于：具有式1结构：
其中，荧光探针具有四取代咪唑环及四级铵盐结构，中间通过疏水性脂肪链连接：由四取代咪唑环、十二烷基链、吡啶环通过成盐连接而成。
2.制备权利要求1所述对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针的方法，其特征在于：包括如下步骤：
a、将4-甲酰基吡啶和苯胺溶于冰醋酸中，于常温下搅拌反应0.5～1.5小时；然后依次加入茴香偶酰和醋酸铵混合均匀，继续加热至120℃回流反应6～12小时；TLC检测反应完全后用水淬灭反应，用氢氧化钠溶液调pH至近中性，观察到有絮状沉淀产生，减压过滤得到滤饼，干燥，柱色谱分离纯化得到式2中间体；
b、将化合物1溶于乙腈中，加入1,12-二溴十二烷，将体系混合均匀，于90℃加热回流，反应6～10小时，结束后，减压蒸馏除掉溶剂，柱色谱分离纯化得到式3中间体；
c、将化合物2溶于吡啶中，体系加热至90℃反应，反应6～10小时，结束后减压蒸馏除掉溶剂，柱色谱分离纯化得到目标化合物；
3.如权利要求2所述制备对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针的方法，其特征在于：聚集诱导荧光发射增强的化合物的合成路线如下：
其中，
化合物1的合成：分别称取4-甲酰基吡啶(107.1mg，1mmol)和苯胺(93.1mg，1mmol)溶于6～8mL冰醋酸中，于常温下搅拌1小时；茴香偶酰(207.2mg，1mmol)和醋酸铵(462.5mg，6mmol)依次加入到反应体系中，在120℃条件下反应过夜，用水淬灭反应，将反应体系倒入200mL冰水中，用0.1mmol/L的氢氧化钠溶液将体系pH调至中性，混合物过滤用水洗三次，真空干燥后硅胶柱层析分离纯化得到产物；
化合物2的合成：称取化合物1(191.5mg，0.44mmol)和1,12-二溴十二烷(145.0mg，0.44mmol)溶解在3mL乙腈溶液中，室温充分搅拌，反应体系在90℃充分回流8小时，TLC板监测反应完全后，减压蒸馏除去溶剂，硅胶柱层析分离得到化合物2；
化合物3的合成：称取化合物2(155.0mg，0.20mmol)溶解在4～5mL吡啶溶液中，室温充分搅拌，反应体系在90℃充分回流8小时，TLC板监测反应完全后，减压蒸馏除去溶剂，硅胶柱层析分离得到目标产物3。
4.一种如权利要求1所述对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针的对溶血磷脂酸特异性识别的荧光分子探针纳米粒的制备方法，其特征在于：将对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针溶于有机溶剂后制备成母液，加入到水溶液中，然后用移液枪吸取母液在超声条件下加入到水溶液中，体系常温搅拌30min，即得对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针纳米粒。
5.根据权利要求4所述的对溶血磷脂酸特异性识别的荧光分子探针纳米粒的制备方法，其特征在于：所述的有机溶剂选自甲醇、乙醇、二甲基甲酰胺、四氢呋喃、二甲亚砜、丙酮、乙腈中的至少一种，所述的水溶液选自纯水、生理盐水、磷酸缓冲液(PBS)、三(羟甲基)氨基甲烷盐酸盐缓冲溶液或4-羟乙基哌嗪乙磺酸缓冲溶液。
6.根据权利要求1所述的对溶血磷脂酸特异性识别的具有聚集诱导发射效应的水溶性荧光分子探针的应用，其特征在于：应用于化学溶液体系、血液中溶血磷脂酸的荧光定量分析检测的水溶性荧光分子探针制备。
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