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Simultaneous Isocratic Determination of Five Catecholamines and Serotonin Under Soft Conditions Using HPLC-UV

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Norepinephrine, Epinephrine, Dopamine, Normetanephrine, Metanephrine, Serotonin.

Scope of The Method

The method is intended for the isocratic determination of five catecholamines (norepinephrine, epinephrine, dopamine, normetanephrine, metanephrine) and serotonin under soft conditions in various pharmaceutical and biological samples using simple isocratic 400 bar HPLC system with a conventional UV detector.

Minimum System Requirements
Solvent Delivery System: Isocratic HPLC pump with 400 bar upper back-pressure limit 
Detector: Single wavelength UV detector
Column Oven: Preferable to ensure retention time stability

Compartible with all HPLC systems: Agilent, Shimadzu, Waters, Thermo, etc.

General Information On The Method
Elution Mode: isocratic, Detection: UV, Actual Analysis Time: 10.0 minutes, Back-pressure: typically < 300 bar, HPLC column manufacturer: HELIX Chromatography, USA >> 


Norepinephrine, Epinephrine, Dopamine, Normetanephrine, Metanephrine, Serotonin, HPLC, Agilent, Shimadzu, Waters, Thermo Scientific

Fig. 1. Isocratic determination of five catecholamines and serotonin. Detection: UV 215 nm.
1. Norepinephrine,  2. Epinephrine, 3. Dopamine, 4. Normetanephrine, 5. Metanephrine, 6. Serotonin.

Norepinephrine, Epinephrine, Dopamine, Normetanephrine, Metanephrine, Serotonin


The method may require soft adjustment to provide an excellent result for a particular sample. Retention and separation selectivity tuning recommendations are given in the application.

        We can fine-tune method performance to meet your requirements upon request.

        We can develop the complete HPLC method based on this application upon request.

Table of contents

Description of The method  5
Name of The Method 5
Scope of The Method  5
Main Characteritics of The Method 5
Minimum System Requirements 5
Analyte(s) 6
Standard HPLC Conditions 6
Typical Chromatogram(s) 7
Suitable HPLC Column(s) 7
Quickstart Steps 8
Mobile Phase Preparation 8
Column Washing And Storage 8
Column Conditioning 8
Testing The Column Performance 8
Fine-Tuning Retention And Selectivity 10


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