Department of Chemistry, College of Science, Princess Nourah bint Abdulrahman University, Riyadh 11671, Saudi Arabia.
Centre for Metabolomics Research, Department of Biochemistry and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool, Liverpool, L69 7ZB, UK.
Analyst. 2023 Apr 11;148(8):1805-1814. doi: 10.1039/d3an00249g.
Paracetamol (also known as acetaminophen) is an over-the-counter (OTC) drug that is commonly used as an analgesic for mild pain, headache, cold and flu. While in the short term it is a safe and effective medicine, it is sometimes used for attempted suicides particularly in young adults. In such circumstances it is important for rapid diagnosis of overdoses as antidotes can be given to limit liver damage from one of its primary metabolites -acetyl--benzoquinone imine (NAPQI). Unfortunately, the demand for rapid and sensitive analytical techniques to accurately monitor the abuse of OTC drugs has significantly risen. Ideally these techniques would be highly specific, sensitive, reproducible, portable and rapid. In addition, an ideal point of care (PoC) test would enable quantitative detection of drugs and their metabolites present in body fluids. While Raman spectroscopy meets these specifications, there is a need for enhancement of the signal because the Raman effect is weak. In this study, we developed a surface-enhanced Raman scattering (SERS) methodology in conjunction with chemometrics to quantify the amount of paracetamol and its main primary metabolites (., paracetamol sulfate, -acetamidophenyl β-D-glucuronide and NAPQI) in water and artificial urine. The enhancement of the SERS signals was achieved by mixing the drug or xenometabolites with a gold nanoparticle followed by aggregation with 0.045 M NaCl. We found that the SERS data could be collected directly, due to immediate analyte association with the Au surface and colloid aggregation. Accurate and precise measurements were generated, with a limit of detection (LoD) of paracetamol in water and artificial urine at 7.18 × 10 M and 2.11 × 10 M, respectively, which is well below the limit needed for overdose and indeed normal levels of paracetamol in serum after taking 1 g orally. The predictive values obtained from the analysis of paracetamol in water and artificial urine were also excellent, with the coefficient of determination () being 0.995 and 0.996, respectively (1 suggests a perfect model). It was noteworthy that when artificial urine was spiked with paracetamol, no aggregating agent was required due to the salt rich medium, which led to spontaneous aggregation. Moreover, for the xenometabolites of paracetamol excellent LoDs were obtained and these ranged from 2.6 × 10 M to 5 × 10 M with paracetamol sulfate and NAPQI having values of 0.934 and 0.892 and for -acetamidophenyl β-D-glucuronide this was slightly lower at 0.6437.
对乙酰氨基酚（也称为扑热息痛）是非处方药（OTC）药物，通常用作轻度疼痛、头痛、感冒和流感的镇痛药。虽然它在短期内是一种安全有效的药物，但有时也会被用于自杀企图，尤其是在年轻成年人中。在这种情况下，快速诊断药物过量非常重要，因为可以使用解毒剂来限制其主要代谢物之一 -乙酰-苯醌亚胺（NAPQI）对肝脏的损害。不幸的是，对能够准确监测 OTC 药物滥用的快速和敏感分析技术的需求显著增加。理想情况下，这些技术应该具有高度特异性、灵敏度、重现性、便携性和快速性。此外，理想的即时护理（PoC）测试应该能够定量检测体液中存在的药物及其代谢物。虽然拉曼光谱符合这些规格，但需要增强信号，因为拉曼效应很弱。在这项研究中，我们开发了一种表面增强拉曼散射（SERS）方法，并结合化学计量学，以定量检测水中和人工尿液中的扑热息痛及其主要初级代谢物（例如，对乙酰氨基酚硫酸盐、-乙酰氨基苯-β-D-葡糖苷酸和 NAPQI）。通过将药物或外源性代谢物与金纳米颗粒混合，然后用 0.045 M NaCl 进行聚集，实现了 SERS 信号的增强。我们发现，由于分析物与 Au 表面的直接结合和胶体聚集，SERS 数据可以直接收集。生成了准确和精确的测量结果，水中扑热息痛的检测限（LoD）和人工尿液中的检测限分别为 7.18×10-7 M 和 2.11×10-7 M，远低于药物过量所需的水平，甚至低于口服 1 克后血清中扑热息痛的正常水平。从水中和人工尿液中分析扑热息痛获得的预测值也非常出色，决定系数（）分别为 0.995 和 0.996（1 表示完美模型）。值得注意的是，当人工尿液中加入扑热息痛时，由于盐含量丰富的介质，不需要聚集剂，这导致自发聚集。此外，对于扑热息痛的外源性代谢物，也获得了极好的 LoD，范围从 2.6×10-7 M 到 5×10-7 M，其中对乙酰氨基酚硫酸盐和 NAPQI 的 值分别为 0.934 和 0.892，而 -乙酰氨基苯-β-D-葡糖苷酸的 值略低，为 0.6437。
