Method Transfer from an Agilent 1200 Series LC to an Agilent 1260 Infinity II LC

Significance of the Topic

Instrument-to-instrument method transfer ensures continuity and consistency of analytical procedures across laboratory platforms. In pharmaceutical quality control and clinical research, transferring an established liquid chromatography (LC) method from legacy systems to modern instruments maintains regulatory compliance, minimizes revalidation efforts, and enhances throughput and reliability.

Objectives and Study Overview

This study demonstrates the seamless transfer of a conventional reversed-phase LC method for six tricyclic antidepressant drugs from an Agilent 1200 Series Quaternary LC to an Agilent 1260 Infinity II LC. Performance metrics such as retention time precision, peak area repeatability, and inter-instrument equivalency were evaluated.

Instrumentation

• Agilent 1200 Series Quaternary LC: Quaternary Pump G1311A, High Performance Degasser G4225A, Autosampler G1329A with thermostat G1330B, Column Compartment G1316A, Variable Wavelength Detector SL G1314C (254 nm).
• Agilent 1260 Infinity II LC: Quaternary Pump G7111B, Vialsampler G7129A with integrated column compartment and sample cooler, Variable Wavelength Detector G7114A (254 nm).
• Column: Agilent ZORBAX Eclipse Plus C18, 4.6 × 150 mm, 5 µm.
• Software: Agilent OpenLAB CDS v2.1.
• Chemicals and Standards: LC-grade methanol and water, 10 mM ammonium bicarbonate buffer pH 8, six TCA hydrochloride salts at 50 µg/mL.

Methodology

The gradient employed solvent A (10 mM ammonium bicarbonate) and solvent B (methanol). Starting at 60 % B, the gradient ramped to 68 % B at 10 min and 90 % B at 20 min, with a 10 min post-run, 1.5 mL/min flow, 40 °C column temperature, and 10 µL injection volume. Samples contained an equimolar mix of amitriptyline, doxepin, imipramine, nortriptyline, protriptyline, and trimipramine in water/methanol (50:50).

Main Results and Discussion

• On the 1200 Series LC, retention times ranged from 3.69 to 13.86 min with RT RSD ≤0.20 % and peak area RSD ≤0.18 %. All adjacent peaks were baseline resolved (resolution ≥2.1).
• A ±2 % change in methanol content shifted RTs by up to –11 % (with more organic) and +15 % (with less organic), highlighting gradient sensitivity.
• After direct method transfer to the 1260 Infinity II LC without parameter adjustment, RTs were 3.76–13.99 min with RT RSD ≤0.06 % and area RSD ≤0.17 %, matching resolution performance.
• Comparison of instruments showed RT deviations ≤1.9 %, confirming proof of equivalency and transfer robustness.

Practical Benefits and Applications

Successful method transfer reduces downtime during instrument upgrades and streamlines method validation. Laboratories benefit from improved precision and reliability of the 1260 Infinity II LC while retaining existing method protocols. This approach supports high-throughput QA/QC of pharmaceutical compounds.

Future Trends and Potential Applications

Advances in ultra-high-pressure LC with Poroshell and core-shell column technologies enable faster separations with lower solvent consumption. Integration of automated sample handling, cloud-based data management, and AI-driven method optimization will further accelerate method transfer and routine analysis across multi-instrument laboratories.

Conclusion

The conventional LC method for tricyclic antidepressants was seamlessly transferred from the Agilent 1200 Series to the Agilent 1260 Infinity II LC. Equivalent retention profiles, precision, and resolution validate the newer platform’s suitability, facilitating efficient laboratory workflows and regulatory compliance.

References

1. Tatsumi M et al. Pharmacological profile of antidepressants and related compounds at human monoamine transporters. European Journal of Pharmacology. 1997;340:249–258.
2. Gillman PK. Tricyclic antidepressant pharmacology and therapeutic drug interactions updated. British Journal of Pharmacology. 2007;151:737–748.