【作者】 肖蓉；

【导师】 吴海龙；

【作者基本信息】 湖南大学 ， 分析化学， 2018， 硕士

【摘要】 化学多维校正方法具有独特的二阶优势,它以“数学分离”来替代或增强传统的物理或化学分离手段,在未知干扰或非三线性因素共存的情况下,依然可以实现复杂基质中多种感兴趣物质的同时定量分析。二阶校正作为一种绿色、高效的分析方法,与现代高阶分析仪器相结合在生命科学、食品、环境和化妆品等领域得到了十分广泛的应用。目前,随着医疗水平的提高和医药改革的持续推进,新药、仿制药的研发和临床试验进程不断加快,开发用于复杂基质中药物分析的新方法和新技术的重要性日益凸显。本文作者采用二阶校正方法结合高阶分析仪器的策略,将其用于严重影响人类健康的抗肿瘤药和作用于心血管系统药物的定量分析,主要包括以下四个方面的内容:第二章:华法林联用阿司匹林可以取得更好的抗凝血效果,但是可能因为两者剂量不合适且调整不及时,而提高临床上的过度出血事件的发生率,在复杂的生物基质中快速、同时测定二者的浓度对临床合理联用药物的指导和药物治疗方案的优化等具有十分重要的意义。本章利用三维荧光结合基于交替三线性分解（ATLD）算法的二阶校正方法,对人血浆和尿液样中华法林和阿司匹林进行了同时定量分析。在不借助复杂样本预处理手段的情况下,尽管分析物之间、分析物与基质之间的荧光光谱都严重重叠,所获得的人血浆和尿液样中两种目标分析物的平均加标回收率依然在91.2-105.4%之间,相应的标准偏差在2.3-8.3%之间,品质因子和日内日间精密度实验结果也表明该方法具有较高的灵敏度、较好的重复性和再现性,是一种简单、高效、绿色、实用的分析方法。并且,采用的荧光检测器获得了较低的检测限(小于0.48 ng mL^-1),能够满足两种联用药物在临床检测中的要求。第三章:沙库必曲联用缬沙坦是首个血管紧张素转化酶受体-脑啡肽酶的双重抑制剂,作为一款重磅抗心衰新药可显著降低心衰死亡率,并改善其他疗效指标,具有非常广阔的市场应用前景。本章采用β-环糊精增敏三维荧光结合二阶校正方法的策略对人体液中的沙库必曲和缬沙坦的含量进行了分析,采用交替归一加权残差（ANWE）算法得到的人血浆和尿液样中沙库必曲和缬沙坦的平均回收率分别为94.0±9.1%,97.6±5.3%和95.2±2.6%,92.7±7.2%,并计算了预测均方根误差（RMSEP）、灵敏度（SEN）、选择性（SEL）以及检测限（LOD）等分析品质因子和统计学参数来评估该方法的性能。结果表明,凭借二阶校正方法的“数学分离”功能和β-环糊精的荧光增敏效果,无需繁琐的样本预处理程序,即使分析体系存在未知干扰以及荧光光谱重叠现象,该方法依然得到了令人满意的结果,具有简单、快速、成本低等优点,有望拓展成为临床新药测定的通用性方法,并为复杂体系中弱荧光物质的测定提供了新思路。第四章:乐伐替尼、帕唑帕尼、阿西替尼、卡博替尼和索拉非尼为靶向血管内皮生长因子受体（VEGFR）的抗癌药,特别适用于晚期肾癌患者的治疗,为了对比它们的治疗效果、安全性和耐受性,优化单独或联用给药方案,降低毒副作用,发展一种简单、通用的测定血浆中抗癌新药浓度的方法对它们的研发和临床试验至关重要。本章将基于交替三线性分解（ATLD）算法的二阶校正方法与HPLC-DAD结合起来,完成了人血浆样中五种抗肾癌药的同时定量分析,分析过程中出现了基线漂移、色谱峰重叠和未知背景干扰等问题,五种目标分析物的加标回收率仍在98.4-109.7%之间,并获得了较好的品质因子和日内日间精密度实验结果。借助ATLD方法的“数学分离”能力,所提出的方法有效地减少了分析时间和有机溶剂的消耗,有望发展成为一种通用性的方法用于复杂生物基质中多种抗癌药的同时定量检测。第五章:川穹为活血化瘀和治疗头痛的常用中药,还可以用于恶性肿瘤和心血管方面疾病的治疗,其活性成分的含量易受气候、产地和加工条件等多种因素的影响。本章将二阶校正方法辅助HPLC-DAD的策略用于两种中药川穹中六种活性成分的含量测定。在简单的梯度洗脱程序下,六种分析物在8 min内全部流出,并利用交替三线性分解（ATLD）算法的二阶优势,成功解决了基线漂移、未知背景干扰和色谱共流出等问题,取得了令人满意的定量结果（平均加标回收率在98.9-110%之间）,精密度实验结果也验证了方法的重复性和再现性。该方法简单、高效、通用性强,对中药材的品质鉴别、质量监管和保证其药效稳定等具有一定的实用价值。更多还原

【Abstract】 The multi-way calibration method can achieve simultaneous quantitative analysis of various components of interest in complex matrices,even in the presence of unknown interferences or non-trilinear factors by utilizing “mathematical separation” to replace or enhance the traditional physical or chemical separation.And this unique property,namely second-order advantage.As a green and efficient analysis method,second-order calibration method combined with high-order analytical instruments has been widely used in the fields of life sciences,food,environment,and cosmetics.Recently,with the improvement of healthcare and continuous advancement of pharmaceutical reform,the progress of research and clinical trials for new drugs and generic drugs are constantly accelerating.Thus,the importance of developing new methods and technologies for drug analysis in complex matrices has become increasingly prominent.The authors adopt the above analysis strategy for the quantitative analysis of anti-tumor and cardiovascular drugs that seriously affect human health,and the main content are as the following:In Chapter 2: Warfarin(WAR)plus low-dose aspirin(ASA)not only might have an additive effect,but also calls for less intensive anticoagulation therapy.However,the risk of bleeding was greatly increased and excessive bleeding events often occur clinically when the doses of both are inappropriate or are not adjusted in time.In this work,a practical method that combines excitation-emission matrix fluorescence with a second-order calibration method based on an alternating trilinear decomposition(ATLD)algorithm was developed in order to simultaneously and directly determine the contents of warfarin and aspirin in human plasma and urine samples,even in the presence of overlapped fluorescence spectra and unknown interferences.With the pre-estimated component number of 4,the obtained average spiked recoveries were 105.4±7.8% and 104.2±8.3% for WAR,96.5±2.8% and 91.2±2.3% for ASA in human plasma and urine samples,respectively.Furthermore,the figures of merits were calculated and inter-and intra-day experiments were also performed that proved the proposed method is of great significance to the monitoring of clinical administration and also being simple sample pre-treatment and low-cost.In Chapter 3: A new compound drug(Sacubitril(SAC)plus valsatan(VAL)),as the first dual inhibitor of angiotensin receptor blocker and neprilysin,was approved by the FDA in 2015.It can not only reduce the mortality rate of patients with heart failure by 20%,but also can significantly improve the efficacy of other indicators compared with enalapril that widely used in clinic,which may make it become the preferred drug for anti-heart failure treatment.In this work,a simple method that combines β-cyclodextrin-enhanced excitation-emission matrix fluorescence with second-order calibration method based on an alternating normalization-weighted error(ANWE)algorithm was developed to simultaneously and directly determine the contents of sacubitril and valsatan in biological fluids,even in the presence of unknown interferences.Without any complex sample pre-treatment steps,the obtained average spiked recoveries were 94.0±9.1% and 95.2±2.6% for SAC,97.6±5.3% and 92.7±7.2% for VAL in human plasma and urine samples,respectively.Moreover,the figures of merit and statistical parameters such as root-mean-square error of prediction(RMSEP),sensitivity(SEN),selectivity(SEL)and limit of detection(LOD)were calculated,which also proved that the proposed method can be developed into a universal method for the analysis of new drugs and provides a strategy for the determination of weak fluorescent substances in complex systems.In Chapter 4: Lenvatinib,pazopanib,axitinib,cabozantinib,and sorafenib that target vascular endothelial growth factor receptor(VEGFR)are particularly suitable for the treatment of patients with advanced kidney cancer.In order to compare their therapeutic efficacy,safety,and tolerability,optimize the dosing regimens,as well as reduce toxicity and side effects,it is of great significance to develop a simple and universal method for the determination of new anti-cancer drugs in plasma.In this chapter,a second-order calibration method based on the alternating trilinear decomposition(ATLD)algorithm combined with high performance liquid chromatography with diode array detector(HPLC-DAD)were proposed for the simultaneous quantitative analysis of five anti-cancer drugs in human plasma sample.Even in the presence of baseline drift,overlapped chromatographic peaks and unknown background interferences and other issues,the spiked recoveries of five target analytes were still between 98.4% and 109.7%,and the results for figures of merit and inter-and intra-day experiments were also satisfactory.Due to the "mathematical separation" capability of the ATLD method,the proposed method effectively reduces the consumption of analysis time and organic solvents.Moreover,it can be developed into a universal method for the simultaneous determination of multiple anti-cancer drugs in complex biological matrices.In Chapter 5: Rhizoma Chuanxiong is a commonly used traditional Chinese medicine for blood circulation and headache treatment.It can also be used for the treatment of cancers and cardiovascular diseases.The content of active ingredients in Rhizoma Chuanxiong is dependent on many factors,such as climate,place of origin,and processing conditions.In this chapter,a chemometrics-assisted high performance liquid chromatography-diode array detection(HPLC-DAD)strategy was applied to the determination of six active ingredients in two Chinese medicine of Chuanxiong.After a simple gradient elution procedure,all six analytes eluted out within 8 minutes.Owing to the second-order advantage of alternating trilinear decomposition(ATLD)method,several problems,such as,baseline drift,unknown background interferences,and chromatographic co-elution were successfully solved.Satisfactory quantitative results were obtained with average spike recoveries between 98.9% and 110%.Furthermore,the results of inter-and intra-day experiments also verified the repeatability and reproducibility of this method.The method is simple,efficient,and universal,which has certain practical value for the quality supervision and ensuring the stability of its medicinal properties.更多还原