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Laser-induced Fluorescence Detector for the Detection of Biogenic Amines

Author: YangRui
Tutor: YangChangLong
School: Qiqihar University
Course: Analytical Chemistry
Keywords: Biogenic amines High Performance Liquid Chromatography Fluoresceinisothi-ocyanate Laser induced Fluorescence Detection
CLC: O657.3
Type: Master's thesis
Year: 2012
Downloads: 99
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Abstract


Laser-induced fluorescence detector (LIFD) is one of the most sensitive detectiontechniques for the detection of chemical and biological samples by high performance liquid chromatography (HPLC) is widely used in micro-column liquidchromatography(Micro-LC) and capillary electrophoresis (CE) and other separation fields. Derivatives ofbiogenic amines and separation process, the first test for the detection sensitivity of the linear relationship of LIFD; Second, the approach of the liquid sample; study derivative temperature, buffer pH, derived reaction time and other factors on biological in the mobile phase, temperature effects and the influence of amine derivatization and chromatographic conditions for biogenic amines in the test beverages.In this paper, fluorescein isothiocyanate (FITC) as a pre-column derivatization reagent on the derivatization of biogenic amines in the beverage, Sinochrom of ODS-BPC18(5μm,4.6mm×200mm) column and high performance liquidchromatography (HPLC) hyphenated techniques with laser-induced fluorescence detector (LIFD), derived products were detected. Derivative product stability, linear range and repeatability for thedetection of indicators, the results show that:(1) The laser-induced fluorescence detector test results showed that the detection limit:7.55×10-15mol/L; sensitivity:1.29×10-12mV/mol/L; line sex: more than104.(2) Experiments using liquid-liquid extraction (LLE) methods for extraction of biogenic amines in beverage samples. The choice of sodium chloride for the extraction of salt, extraction with ethyl acetate under alkaline conditions of pH=11,achieved goodresults.(3) The derivative conditions are optimized, and biogenic amines were measured atpH=9.0,18h under the conditions of constant temperature50°C under the derivatizationreaction to fluorescein isothio-cyanate (FITC) as derivative reagent. And successfully used forthe qualitative separation of biogenic amines drinks, the chromatographic conditions: mobilephase was methanol: water=7:3,flow rate1mL/min.

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