Development and Verification of a Generic Agilent 1260 Infinity II Binary HPLC Method

Importance of the Topic

The simultaneous quantification of multiple active ingredients in OTC cold and cough medications is critical for quality control and regulatory compliance. Rapid and robust analytical methods support efficient manufacturing workflows and ensure product safety and efficacy.

Objectives and Study Overview

This note describes the development and verification of a generic reversed phase HPLC method using an Agilent 1260 Infinity II system coupled with an InfinityLab Poroshell 120 EC C8 column. The goal was to achieve baseline separation of seven active ingredients within a runtime of five minutes, improving throughput compared with existing assays.

Methods and Instrumentation

• System Agilent 1260 Infinity II binary pump, multisampler, multicolumn thermostat, diode array detector
• Column InfinityLab Poroshell 120 EC C8 2.1 x 150 mm 2.7 μm
• Mobile phase A 0.1 percent trifluoroacetic acid in water; mobile phase B acetonitrile methanol 75:25 v/v with 0.1 percent TFA
• Gradient from 15 to 100 percent B over five minutes with a three-minute post run
• Flow rate 0.35 mL/min; injection volume 5 μL; detection at 280 nm; column temperature 30 °C

Main Results and Discussion

The optimized method achieved baseline separation of acetaminophen, dextromethorphan, doxylamine, phenylephrine, guaifenesin, caffeine, and aspirin within five minutes. Linearity over 30 to 140 percent of target concentration yielded correlation coefficients above 0.999. Precision RSD values were below 0.2 percent and no interferences were observed. Robustness testing with flow rate and temperature variations showed minimal impact on resolution.

Benefits and Practical Applications

• High throughput with run time under five minutes
• Low solvent consumption about 1.75 mL per injection
• Generic method applicable to a broad range of cold and cough formulations
• Ideal for routine quality control in pharmaceutical and biopharma labs

Future Trends and Potential Applications

Advances in superficially porous particle technology and smaller particle sizes will allow even faster analyses with higher efficiency. Coupling to mass spectrometry can expand the method to trace impurity profiling. Further automation and AI driven data processing will streamline method transfer and compliance.

Conclusion

A fast, cost effective generic RP HPLC method for simultaneous analysis of seven OTC actives was developed and verified. The approach offers significant time savings, low solvent use, and robust performance for routine pharmaceutical quality control.

References

1. Dongala T et al Development and Validation of a Generic RP HPLC Method Biomed Chromatogr 2019
2. Burge LJ and Raches DW A Rapid HPLC Assay for Combination Analgesics J Liq Chrom Relat Technol 2003
3. Dong Y et al Separation and Determination of Multiple Cold Medication Actives J Pharm Biomed Anal 2005
4. Heydari R A New HPLC Method for Simultaneous Determination of Four Actives Anal Lett 2008