top of page
    Ready-to-Use HPLC Applications

Simultaneous specific determination of five fluoroquinolone antibiotics using isocratic HPLC-UV

Document #030120-03. Revision 01, 01 March 2020



Sparfloxacin, Enrofloxacin, Ofloxacin, Ciprofloxacin, Norfloxacin.

Scope of The Method

The method is intended for the highly selective determination of five fluoroquinolone antibiotics (sparfloxacin, enrofloxacin, ofloxacin, ciprofloxacin, norfloxacin) in various pharmaceutical and biological samples using simple isocratic 400 bar HPLC system with a conventional UV detector.
 The method is capable to determine fluoroquinolone antibiotics in complex matrices that contain any neutral or basic matrix compounds.

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: 8.0 minutes, Back-pressure: typically < 100 bar, HPLC column manufacturer: Nouryon, Sweden >>

Sparfloxacin, Enrofloxacin, Ofloxacin, Ciprofloxacin, Norfloxacin, HPLC, Agilent, Shimadzu, Waters, Thermo Scientific

Fig. 1. Specific determination of five fluoroquinolone antibiotics.

Detection: UV 290 nm. 1. Sparfloxacin, 2. Enrofloxacin, 3. Ofloxacin, 4. Ciprofloxacin, 5. Norfloxacin.

Sparfloxacin, Enrofloxacin, Ofloxacin, Ciprofloxacin, Norfloxacin


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 9


bottom of page