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Simultaneous Isocratic Determination of Eight Drugs in Cough & Cold Preparations That Include Acetaminophen and Caffeine Using HPLC-UV

Document #030820-07. Revision 01, 08 March 2020



Phenylephrine, Acetaminophen, Pseudoephedrine, Metamizole, Codeine, Guaifenesin, Caffeine, Phenobarbital.

Scope of The Method

The method is intended for the isocratic determination of eight drugs (phenylephrine, acetaminophen, pseudoephedrine, metamizole, codeine, guaifenesin, caffeine, phenobarbital) in in cough & cold preparations 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: 15 minutes at 2mL/min; 30 minutes at 1mL/min, Back-pressure: typically < 300 bar at 2mL/min; < 150 bar at 1mL/min, HPLC column manufacturer: Nacalai Tesque, Japan >>


Phenylephrine, Acetaminophen, Pseudoephedrine, Metamizole, Codeine, Guaifenesin, Caffeine, Phenobarbital, HPLC, Agilent, Shimadzu, Waters, Thermo Scientific

Figure 1. Isocratic determination of eight drugs in cough & cold preparations. Detection: UV 215 nm + 280 nm.
1. Phenylephrine, 2. Acetaminophen, 3. Pseudoephedrine, 4. Metamizole, 5. Codeine, 6. Guaifenesin, 7. Caffeine, 8. Phenobarbital.


Phenylephrine, Acetaminophen, Pseudoephedrine, Metamizole, Codeine, Guaifenesin, Caffeine, Phenobarbital


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
Demonstration of Specificity 8
Quickstart Steps 9
Mobile Phase Preparation 9
Column Washing And Storage 9
Column Conditioning 9
Testing The Column Performance 9
Fine-Tuning Retention And Selectivity 10


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