کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1200075 1493573 2014 12 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Novel and sensitive reversed-phase high-pressure liquid chromatography method with electrochemical detection for the simultaneous and fast determination of eight biogenic amines and metabolites in human brain tissue
ترجمه فارسی عنوان
با استفاده از روش کروماتوگرافی مایع با فشار بالا و با حساسیت الکتروشیمیایی ریز و حساس به منظور تشخیص همزمان و سریع هشت آمین و متابولیت های بیوگرافی در بافت مغز انسان
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• Design of experiment utilization for RP-HPLC-ECD method optimization.
• Method validation according to SFSTP guidelines.
• Simultaneous sensitive determination of eight biogenic amines and metabolites.
• Application in post mortem human brain tissue.

A fast and simple RP-HPLC method with electrochemical detection (ECD) and ion pair chromatography was developed, optimized and validated in order to simultaneously determine eight different biogenic amines and metabolites in post-mortem human brain tissue in a single-run analytical approach. The compounds of interest are the indolamine serotonin (5-hydroxytryptamine, 5-HT), the catecholamines dopamine (DA) and (nor)epinephrine ((N)E), as well as their respective metabolites, i.e. 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), 5-hydroxy-3-indoleacetic acid (5-HIAA) and 3-methoxy-4-hydroxyphenylglycol (MHPG). A two-level fractional factorial experimental design was applied to study the effect of five experimental factors (i.e. the ion-pair counter concentration, the level of organic modifier, the pH of the mobile phase, the temperature of the column, and the voltage setting of the detector) on the chromatographic behaviour. The cross effect between the five quantitative factors and the capacity and separation factors of the analytes were then analysed using a Standard Least Squares model. The optimized method was fully validated according to the requirements of SFSTP (Société Française des Sciences et Techniques Pharmaceutiques). Our human brain tissue sample preparation procedure is straightforward and relatively short, which allows samples to be loaded onto the HPLC system within approximately 4 h. Additionally, a high sample throughput was achieved after optimization due to a total runtime of maximally 40 min per sample. The conditions and settings of the HPLC system were found to be accurate with high intra and inter-assay repeatability, recovery and accuracy rates. The robust analytical method results in very low detection limits and good separation for all of the eight biogenic amines and metabolites in this complex mixture of biological analytes.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1353, 1 August 2014, Pages 28–39
نویسندگان
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