化学
色谱法
醋酸
组胺
尿
样品制备
亲水作用色谱法
基质(化学分析)
咪唑
稀释
高效液相色谱法
生物化学
药理学
医学
热力学
物理
作者
Maxim Nelis,Lisse Decraecker,Guy E. Boeckxstaens,Patrick Augustijns,Deirdre Cabooter
出处
期刊:Talanta
[Elsevier]
日期:2020-12-01
卷期号:220: 121328-121328
被引量:21
标识
DOI:10.1016/j.talanta.2020.121328
摘要
An LC-MS/MS method was developed enabling the separation and quantification of histamine and its main metabolites (imidazole acetaldehyde, imidazole acetic acid, methyl imidazole acetic acid, methyl histamine, acetyl histamine) in urine samples. A fast separation was achieved in 10 min on two HILIC columns connected in series by adopting a linear gradient followed by an isocratic hold. The sample preparation consisted of a simple dilution step wherein 10 μL of urine was diluted with acetonitrile (ACN) to a final volume comprising 95% ACN. For methyl imidazole acetic acid, an additional dilution step was incorporated due to its high natural levels. Hereafter, the samples were stored at −20 °C and centrifuged prior to injection. Matrix matched calibrators were unavailable due to the endogenous occurrence of the compounds of interest. The occurrence of matrix effects and the lack of labeled internal standards prompted the use of the standard addition method as a viable alternative to solvent calibration. The validation of the method entailed matrix effects, accuracy and precision and was performed in compliance with the recent guidelines on endogenous compounds issued by the International Conference of Harmonization (ICH). The method was then adopted for the quantification of histamine and its metabolites in human urine samples collected from healthy volunteers and patients suffering from gastrointestinal discomfort.
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