作者：刘书瑶1, 付晓芸1, 雍智全2, 张传维1, 钟卓伶1, 黄骏3, 徐小平1, 杨明4
作者单位：1. 四川大学华西药学院，四川 成都 610041;
2. 广州安好医药科技有限公司，广东 广州 510700;
3. 成都科美迪检测检验有限公司，四川 成都 610041;
4. 四川体育职业学院康复科研中心，四川 成都 610041
关键词：碳点;DX1002;荧光猝灭;特异性;含量测定
摘要：
针对肿瘤血管抑制剂DX1002的含量测定，构建一种基于新型纳米碳点材料（CDs）的特异性荧光定量方法。以柠檬酸和尿素为原料，热解法制得碳点（CACCDs），经TEM、IR、UV/Vis、Flu等方法对其进行结构表征。内滤光效应下碳点荧光可被DX1002特异性定量猝灭，CACCDs浓度为250 μg/mL时，于λex/λem=400/530 处测定猝灭前后的荧光差值(ΔF)。结果表明，DX1002浓度在2.5~75 μg/mL范围内与ΔF具良好线性关系（r2=0.9988），检出限为1.16 μg/mL，平均加标回收率为101.7%（RSD=2.08%），常见细胞阳离子、微量金属离子、糖类、氨基酸等潜在共存物质及有关物质对测定无干扰，DX1002含量测定的结果（101.3%±1.33%）与HPLC法测定结果（99.4%±1.19%）基本吻合（P>0.05），但较HPLC分析时间快约75倍。该方法快速、灵敏、特异性强，可为建立DX1002高通量特异性体内分析方法提供体外定量的依据。
Construction of specific quantitative method for tumor vascular inhibitor DX1002 based on carbon nanoparticles
LIU Shuyao1, FU Xiaoyun1, YONG Zhiquan2, ZHANG Chuanwei1, ZHONG Zhuoling1, HUANG Jun3, XU Xiaoping1, YANG Ming4
1. West China School of Pharmacy, Sichuan University, Chengdu 610041, China;
2. Guangzhou Anhao Pharmaceutical Technology Co., Ltd., Guangzhou 510700, China;
3. Chengdu COL-MED Inspection & Testing Co., Ltd., Chengdu 610041, China;
4. Sichuan Sports College Rehabilitation Research Center, Chengdu 610041, China
Abstract: LA fluorescence method based on carbon dots (CDs), a novel carbon nanoparticles was developed for the quantitative determination of DX1002. Carbon dots (CACCDs) were prepared from citric acid and urea by pyrolysis, and their structures were characterized by TEM, IR, UV/Vis and Flu. The fluorescence of carbon dots can be specifically quenched by DX1002 under the IFE (internal filter effect), and the fluorescence difference (ΔF) was measured at λex/λem=400/530 when the concentration of CACCDs was 250 μg/mL. The results showed that the fluorescence difference (ΔF) at λex/λem=400/530 nm had a good linear relationship with DX1002 concentration in the range of 2.5-75 μg/mL (r2=0.9988). The detection limit was 1.16 μg/mL and the average recovery was 101.7% (RSD=2.08%). Common cell cations, trace metal ions, sugars, amino acids and other potential coexisting substances and related substances did not interfere with the determination. The results of the determination of DX1002 by HPLC method (99.4%±1.19%) were similar (P>0.05) to this method (101.3%±1.33%), but the analysis time was about 75 times faster than that of HPLC. The method is rapid, sensitive and specific, which can provide quantitative basis for the establishment of DX1002 high-throughput and specific in vivo analysis method.
Keywords: carbon dots;DX1002;fluorescence quenching;specificity;assay
2021, 47(2):68-73,80 收稿日期: 2020-08-20;收到修改稿日期: 2020-09-24
基金项目: 四川省科技计划项目（2020YFS0260）；广州经济技术开发区创新创业领军人才项目（2020-L019）
作者简介: 刘书瑶(1997-),女,陕西汉中市人,硕士研究生,专业方向为药物分析
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