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Updated: Feb 10, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
[Determination of 5-hydroxytryptamine,Adrenalin and Noradrenalin in Human Plasma by Pre-column Derivatization
Jin Xiang1, Qin Yu1, Mao-Zhi Liang1
1Department of Clinical Pharmacology,West China Hospital,Sichuan University,Chengdu 610041,China.
Objective:
To establish a method for the determination of 5-hydroxy tryptamine (5-HT),adrenalin (AD) and noradrenalin (NA) in human plasma using pre-column derivatization HPLC-MS/MS. Methods Derivatives of the plasma samples were produced with dansyl chloride under pH 10.5,and then extracted and enriched with ethyl acetate. The detected chemicals were separated using Ultimate C₁₈ (50 mm×4.6 mm,5 μm) with acetonitrile-water-formic acid=95∶5∶0.05 (V∶V∶V) at 0.2 mL/min. The HPLC-MS//MS system was operated under a multiple reaction monitoring mode (MRM) using the electrospray ionization technique in positive mode.
Results:
Linear range of the calibration curve appeared in 250-2.5 ng/mL. The method had a less than 9% intra- and inter-assay relative standard deviation (RSD),95.44%-109.71% recoveries,and 4.86%-12.81% matrix effects.
Conclusion:
This method is simple,accurate,and suitable for detection of 5-HT,AD and NA in human plasma.
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