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一种用于定量测定尿多巴胺水平的快速、精确且灵敏的液相色谱-串联质谱法——一种简单的液-液萃取技术。

A Rapid, Precise, and Sensitive LC-MS/MS Method for the Quantitative Determination of Urinary Dopamine Levels a Simple Liquid-liquid Extraction Technique.

作者信息

Battal Dilek, Aktaş Süküroğlu Ayça, Alkaş Fehmi Burak, Ünlüsayın İrfan

机构信息

Mersin University Faculty of Pharmacy, Department of Pharmaceutical Toxicology, Mersin, Turkey.

Gazi University Faculty of Pharmacy, Department of Pharmaceutical Toxicology, Ankara, Turkey.

出版信息

Turk J Pharm Sci. 2021 Dec 31;18(6):761-769. doi: 10.4274/tjps.galenos.2021.60486.

Abstract

OBJECTIVES

Dopamine (DA) is a prominent biochemically complex neurotransmitter and immunomodulator. The quantification of DA could contribute to a better understanding of how endocrine system, cardiovascular and renal functions are regulated. The study aims to develop a rapid, precise, and extremely sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for routine clinical quantification of DA in urine.

MATERIALS AND METHODS

Urine samples were extracted one simple and rapid liquid-liquid extraction technique; then analyzed using a sensitive LC-MS/MS method developed by multiple reaction monitoring mode.

RESULTS

DA and internal standard (IS) retention durations were found to be 2.28 min and 2.24 min, respectively. The mean extraction recovery of DA and DA-IS in urine was above 95.62%. DA calibration curve in urine was linear (r≥0.998) ranging from 20 ng/mL to 1000 ng/mL. The maximum intra-day and inter-day precisions were 5.87 and 2.81, respectively and coefficients of variation were 10.55% and 7.57%, respectively.

CONCLUSION

A rapid, precise, sensitive and quantitative LC-MS/MS detection of DA without the use of derivatization, evaporation, reconstitution and ion-pairing reagents has been developed with a simple and non-invasive sample technique for clinical laboratory applications, basic neuroscience research and drug development studies.

摘要

目的

多巴胺(DA)是一种重要的、生化性质复杂的神经递质和免疫调节剂。对DA进行定量有助于更好地理解内分泌系统、心血管和肾脏功能是如何调节的。本研究旨在开发一种快速、精确且极其灵敏的液相色谱-串联质谱(LC-MS/MS)方法,用于尿液中DA的常规临床定量分析。

材料与方法

采用一种简单快速的液液萃取技术对尿液样本进行萃取;然后使用通过多反应监测模式开发的灵敏LC-MS/MS方法进行分析。

结果

发现DA和内标(IS)的保留时间分别为2.28分钟和2.24分钟。尿液中DA和DA-IS的平均萃取回收率高于95.62%。尿液中DA校准曲线在20 ng/mL至1000 ng/mL范围内呈线性(r≥0.998)。日内和日间最大精密度分别为5.87和2.81,变异系数分别为10.55%和7.57%。

结论

已开发出一种快速、精确、灵敏且定量的LC-MS/MS检测方法,无需使用衍生化、蒸发、复溶和离子对试剂,采用简单且无创的样本技术,可用于临床实验室应用、基础神经科学研究和药物开发研究。

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