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

Significance of the Topic

In the pharmaceutical industry, verified chromatographic methods underpin quality control and regulatory compliance. Transferring a United States Pharmacopoeia (USP) method from legacy LC systems to modern platforms ensures continuity of validated procedures while leveraging advances in speed, efficiency, and solvent economy.

Objectives and Scope

This study demonstrates the seamless transfer of the USP monograph method for related compounds of abacavir sulfate from an Agilent 1100 Series Quaternary LC to an Agilent 1260 Infinity III LC. Additionally, it illustrates the translation of that method to UHPLC conditions using an InfinityLab Poroshell column, within permitted USP adjustments.

Methodology and Instrumentation

The USP method employs a reversed-phase C18 column (3.9×150 mm, 5 µm), mobile phases A (0.05% TFA in water) and B (methanol/water 17:3), a linear gradient from 5% to 90% B over 35 min, flow 1.0 mL/min, column temperature 30 °C, UV detection at 254 nm, 20 µL injection, and a 50 min runtime. System suitability requires resolution ≥1.5 between abacavir and trans-abacavir.

Used Instrumentation

• Agilent 1100 Series Quaternary LC: pump G1311A, degasser G1322A, autosampler G1313A, column compartment G1316A, DAD G1315B.
• Agilent 1260 Infinity III LC: pump G7111B, vialsampler G7129C, multicolumn thermostat G7116A, DAD WR G7115A.
• Columns: ZORBAX StableBond C18 (4.6×150 mm, 5 µm) and InfinityLab Poroshell 120 SB-C18 (3.0×75 mm, 2.7 µm).
• Software: Agilent OpenLAB CDS v2.7.
• Chemicals: LC-grade methanol, water, TFA; USP abacavir related compounds standard.

Results and Discussion

Performance checks with an LC performance standard showed excellent precision and peak shape on both systems. The USP method on the 1100 Series produced resolution of 2.6 and retention time RSDs <0.05%. Transfer to the 1260 Infinity III with a 4.6×150 mm column (flow 1.39 mL/min, injection 27.8 µL) preserved resolution (2.6) and precision (<0.1% RT RSD). A further transfer to the 3.0×75 mm, 2.7 µm Poroshell column (flow 1.10 mL/min, injection 5.92 µL) reduced runtime to 13.5 min and solvent use by 78.6%, while boosting resolution to 3.1 and maintaining RSDs below 0.05% for retention times.

Benefits and Practical Applications

• Maintains compliance with USP system suitability criteria.
• Enables method continuity across generations of LC hardware.
• Shortens analysis time by up to 70% and cuts solvent consumption by nearly 80% in UHPLC mode.
• Reduces per-sample cost and enhances laboratory throughput.

Future Trends and Applications

Ongoing developments in superficially porous particle columns and higher-pressure LC platforms will further accelerate analyses. Automated method-transfer tools and advanced data-processing algorithms will simplify validation across laboratories and instruments. Integration with real-time data analytics promises dynamic method adjustment for continuous manufacturing and quality by design.

Conclusion

The USP abacavir related-compounds method was successfully migrated from Agilent 1100 Series to 1260 Infinity III LC and translated to UHPLC conditions. This workflow preserves analytical performance while significantly improving efficiency and sustainability.

References

1. USP general chapter <1224> Transfer of Analytical Procedures. Official prior to 2013; accessed November 14, 2024.
2. USP general chapter <1226> Verification of Compendial Procedures. Official as of December 1, 2019; accessed November 14, 2024.
3. USP Monograph on Abacavir Sulfate. Official as of May 1, 2020; accessed January 10, 2025.
4. Agilent Technologies Technical Overview, Method Transfer from an 1100 Series Quaternary LC to a 1260 Infinity III LC. Publication 5994-8031EN, 2025.
5. USP general chapter <621> Chromatography. Official as of December 1, 2024; accessed January 10, 2025.