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

Significance of the Topic

The reliable migration of chromatographic methods between instruments is vital for maintaining continuity in regulated environments such as pharmaceuticals, food safety, and quality control laboratories. Demonstrating equivalency ensures that analytical data remain consistent and compliant with industry standards when transitioning from legacy equipment to modern systems.

Objectives and Study Overview

This work evaluates the transfer of an HPLC method for eight polyphenolic compounds from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity II LC. Key performance parameters—resolution, retention time precision, and peak area reproducibility—are compared across both systems. Additionally, the method is adapted to UHPLC conditions using Agilent InfinityLab Poroshell columns to assess gains in speed and solvent economy.

Methodology and Instrumentation

The original separation was performed on an Agilent 1100 Series Quaternary LC equipped with:

• Quaternary Pump (G1311A)
• Degasser (G1379A)
• Autosampler (G1313A)
• Thermostatted Column Compartment (G1316A)
• Diode Array Detector (G1315B) with 10 mm flow cell

The target system was an Agilent 1260 Infinity II LC in two configurations:

• Quaternary Pump (G7111B), Vialsampler with 6 µL heat exchanger (Option #66), and Variable Wavelength Detector (G7114A)
• Vialsampler with 3 µL heat exchanger (Option #063)

Chromatographic conditions:

• Column: ZORBAX SB-C18, 4.6 × 150 mm, 5 µm; mobile phase A: 0.1 % formic acid/water, B: 0.1 % formic acid/acetonitrile; gradient 10–30 % B over 12 min; flow 1.5 mL/min; 30 °C; detection 280 nm.
• UHPLC upgrade: InfinityLab Poroshell 120 SB-C18, 2.1 × 150 mm, 2.7 µm; flow 0.6 mL/min; gradient shortened to 5.4 min; total run time 7 min; 30 °C; detection 280 nm.

All data were acquired and processed using Agilent OpenLAB CDS v2.1.

Main Results and Discussion

On the 1100 Series, six consecutive runs yielded retention time RSDs below 0.14 % and area RSDs under 0.10 %. Transferring to the 1260 Infinity II with the 6 µL heat exchanger maintained similar precision (< 0.08 % RT RSD, < 0.12 % area RSD). The 3 µL exchanger also delivered equivalent performance. Retention time shifts after transfer were within ±0.02 min (±1.2 %) compared to the original system, and inter‐exchanger shifts did not exceed ±0.03 min (±0.7 %).

The UHPLC method achieved baseline separation in half the time, reducing analysis duration by 52 % and solvent usage by over 80 %. Precision remained high (RT RSD < 0.07 %, area RSD < 0.23 %).

Benefits and Practical Applications

These findings validate that legacy HPLC methods can be seamlessly migrated to advanced LC platforms without compromising data quality. The ability to retrofit methods onto UHPLC‐compatible columns offers significant throughput gains and cost savings in routine polyphenol analysis and can be extended to diverse compound classes.

Future Trends and Opportunities

As instrumentation evolves, software emulation tools like ISET will further simplify method migration across vendors. Miniaturized flow paths and enhanced detection technologies may enable even faster separations with minimal sample and solvent requirements. Integration with automated workflows and real‐time data analytics will bolster high‐throughput applications in pharmaceutical development and food safety screening.

Conclusion

The study confirms that the Agilent 1260 Infinity II LC matches or exceeds the performance of the Agilent 1100 Series Quaternary LC for polyphenol separations. Both heat exchanger configurations in the new system deliver equivalent precision and minimal retention time shifts. Transition to UHPLC conditions using Poroshell columns substantially accelerates analysis while maintaining quality metrics, demonstrating a robust pathway for method modernisation.

References

• Krieger S. Method Transfer from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity II LC: Proof of Equivalency for the Analysis of Antihistaminic Drugs. Agilent Technologies Technical Overview, 5991-6914EN, 2016.
• Agilent Technologies. Agilent 1290 Infinity with ISET User Manual. Publication G4220-90314, 2015.