Method Transfer from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity II LC

Importance of the topic

Instrument-to-instrument transfer of liquid chromatography methods is a key challenge in pharmaceutical and quality control laboratories. Adapting validated methods from legacy systems to modern instruments ensures continuity of data quality, regulatory compliance, and operational efficiency while reducing method revalidation efforts.

Aims and overview of the study

This study demonstrates the transfer of a conventional HPLC method for the analysis of antihistaminic drugs from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity II LC. It evaluates equivalency in retention time and resolution, and further adapts the method to UHPLC conditions optimized separately for enhanced resolution and for rapid analysis.

Methodology and instrumentation

The conventional method employed a ZORBAX SB-C18 column (4.6×150 mm, 5 µm) with a gradient from 10% to 55% acetonitrile in 25 mM phosphate buffer (pH 3) over 25 min at 1.5 mL/min. UHPLC adaptations used Poroshell 120 EC-C18 columns (3.0×150 mm, 2.7 µm for resolution; 3.0×50 mm, 2.7 µm for speed) under similar gradient conditions compressed to 13.3 min and 2.13 min, respectively.

Used instrumentation

• Agilent 1100 Series Quaternary Pump, Degasser, Autosampler, Thermostatted Column Compartment, Diode Array Detector
• Agilent 1260 Infinity II LC with Quaternary Pump, Vialsampler with column compartment and cooler, Diode Array Detector WR
• Columns: ZORBAX SB-C18 (4.6×150 mm, 5 µm); Poroshell 120 EC-C18 (3.0×150 mm, 2.7 µm; 3.0×50 mm, 2.7 µm)
• Software: Agilent OpenLAB CDS Version 2.1
• Chemicals: LC-grade acetonitrile, ultrapure water, 25 mM potassium dihydrogen phosphate buffer (pH 3), antihistaminic standards (tripelenamine, chlorpheniramine, tetracaine, promethazine)

Main results and discussion

Equivalency between the 1100 Series and 1260 Infinity II LC was confirmed with a maximum retention time deviation of –1.7% and equivalent or slightly improved resolution. UHPLC adaptation for resolution increased the critical peak resolution from 2.3 to 5.4 while reducing runtime by 47% and solvent consumption by 57%. The rapid UHPLC method achieved a 91% reduction in analysis time and 86% solvent savings with resolution maintained above 2.9.

Benefits and practical applications

This method transfer approach enables laboratories to migrate established HPLC assays to modern UHPLC platforms with minimal revalidation effort. It delivers greater throughput, higher resolution, and lower solvent costs, supporting efficient QA/QC workflows in pharmaceutical development and production.

Future trends and applications

Continued evolution of UHPLC systems with higher pressure capability, smaller particle and core-shell columns, and integrated automation will further accelerate analysis speed and resolution. Green chromatography initiatives will drive solvent reduction and alternative mobile phases. Advanced software platforms will simplify method transfer and regulatory compliance reporting.

Conclusion

The transfer from an Agilent 1100 Series Quaternary LC to a 1260 Infinity II LC delivers equivalent analytical performance for antihistaminic drugs. UHPLC adaptations on Poroshell 120 columns achieve significant improvements in resolution, speed, and solvent efficiency, underscoring the value of modern LC technology for pharmaceutical analysis.

References

1. Agilent Technologies. Method Transfer from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity II LC, Application Note 5991-6914EN, 2016.
2. Agilent Technologies. 1290 Infinity with ISET, User Manual G4220-90314, 2015.
3. Panula P, et al. International Union of Basic and Clinical Pharmacology. XCVIII. Histamine Receptors. Pharmacol. Rev. 2015;67:601–655.