Seamless transfer of elution gradients from Agilent 1100/1200 Series LCs to an Agilent 1290 Infinity LC using ISET
Technical notes | 2012 | Agilent TechnologiesInstrumentation
Method transfer between legacy HPLC and modern UHPLC platforms is critical for highly regulated laboratories. Differences in system delay volumes and gradient mixing can lead to altered retention times and peak resolution, necessitating method adaptation and revalidation.
This technical overview evaluates Agilent Intelligent System Emulation Technology (ISET) on the 1290 Infinity LC System for reproducing elution gradients from Agilent 1100 Series Quaternary LC systems without modifying original methods.
Tracer experiments employed uracil (10 mg/L) in methanol, with water and methanol mobile phases. A restriction capillary column at 30 °C and diode array detection at 254 nm were used. Instrumentation details:
Six gradient profiles were tested: three containing mixed slopes and isocratic steps, and three linear gradients of 3, 10, and 20 minutes. Overlays of recorded tracer responses demonstrated curve correlations above 99.9% between the 1100 Series and the 1290 Infinity LC with ISET, even for ultrafast slope changes. Retention times and resolution remained consistent across all runs.
Potential developments include extending ISET compatibility to third-party systems, refining gradient-shaping algorithms, and integrating AI-driven optimization for automated method transfer.
Agilent ISET enables near-perfect emulation of HPLC gradient profiles on the 1290 Infinity LC, ensuring reliable retention time and resolution transfer and streamlining method migration.
HPLC
IndustriesManufacturerAgilent Technologies
Summary
Significance of the Topic
Method transfer between legacy HPLC and modern UHPLC platforms is critical for highly regulated laboratories. Differences in system delay volumes and gradient mixing can lead to altered retention times and peak resolution, necessitating method adaptation and revalidation.
Objectives and Study Overview
This technical overview evaluates Agilent Intelligent System Emulation Technology (ISET) on the 1290 Infinity LC System for reproducing elution gradients from Agilent 1100 Series Quaternary LC systems without modifying original methods.
Methodology and Instrumentation
Tracer experiments employed uracil (10 mg/L) in methanol, with water and methanol mobile phases. A restriction capillary column at 30 °C and diode array detection at 254 nm were used. Instrumentation details:
- Agilent 1100 Series Quaternary LC: pump G1311A, autosampler G1313A, column compartment G1316A, diode array detector G1315B
- Agilent 1290 Infinity LC with ISET: pump G4220A, autosampler G4226A, thermostat G1330B, column compartment G1316C, diode array detector G4212A
- Software: Agilent ChemStation revision C.01.03 and ISET revision 1.0
Main Results and Discussion
Six gradient profiles were tested: three containing mixed slopes and isocratic steps, and three linear gradients of 3, 10, and 20 minutes. Overlays of recorded tracer responses demonstrated curve correlations above 99.9% between the 1100 Series and the 1290 Infinity LC with ISET, even for ultrafast slope changes. Retention times and resolution remained consistent across all runs.
Benefits and Practical Applications
- Seamless migration of legacy methods without instrument or method modifications
- Minimized validation and development effort in regulated environments
- Preserved chromatographic performance when upgrading to UHPLC technology
Future Trends and Opportunities
Potential developments include extending ISET compatibility to third-party systems, refining gradient-shaping algorithms, and integrating AI-driven optimization for automated method transfer.
Conclusion
Agilent ISET enables near-perfect emulation of HPLC gradient profiles on the 1290 Infinity LC, ensuring reliable retention time and resolution transfer and streamlining method migration.
References
- Agilent 1290 Infinity LC with Intelligent System Emulation Technology, Publication 5990-8670EN, 2011
- Agilent Publication 5990-9545EN, 2012
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