1290 Infinity II 制备型开放式馏分收集器的性能特点 - 纯化组分的纯度及回收率

Abstract 

The Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector was developed and optimized to enable efficient fraction collection with extended capacity and flexibility while providing the highest recovery and purity of the collected fractions. This Technical Overview demonstrates the performance of the 1290 Infinity II Preparative Open-Bed Fraction Collector when peak-based fraction collection is used. Using only the ultraviolet (UV) detection signal or a combination of UV and mass-selective detection (MSD) signals as a fraction trigger, purity and recovery of collected fractions are determined. Furthermore, the peak-based, collecting time slices fraction mode is discussed from a quantitative point of view.

Introduction 

Fraction collection in preparative-scale high-performance liquid chromatography (HPLC) can be challenging. Along with a suitable solvent gradient that separates the compounds of interest from unwanted impurities, the delay between the detector and the fraction collector plays a crucial role in the optimization process. Pure fractions with high recovery can only be collected with accurate setting of the time needed for a substance to migrate from the detector cell to the fraction nozzle. The Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector1 has been optimized in this respect to deliver efficient and flexible fraction collection with high purity and recovery. A built-in delay sensor allows precise calibration of the delay between each detector and the fraction valve. The calibration procedure does not require any hardware modifications, and needs to be done only once on UV-based systems; when the flow rate changes, the delay time is recalculated automatically, and adjusted accordingly. On systems including an MSD, the delay time is not only dependent on the flow rate, but also on the split ratio of the main flow and the make-up flow. 

Exchangeable tubing kits are available with two different inside diameters to fit a wide range of flow rates. They can be cut in length to minimize peak dispersion between the detector and fraction collector. Fraction tubes, available in four different diameters (12, 16, 25, and 30 mm) and two lengths (100 and 150 mm), offer the right choice for any flow rate and fraction size. Carefully choosing fraction tubes is recommended to ensure short pathways of the fraction nozzle between two fractions, or to provide enough space for large fractions without the need to split them into multiple tubes.

Eight peak- and time-based modes, including two recovery modes, are available for fraction collection triggering. The different fraction modes, along with their typical applications, are described in detail in another Technical Overview2 . This document focuses on the recovery and purity of fractions that were collected in peak-based mode. Fraction collection is triggered by the UV signal only, or by a logical AND combination of UV and MSD signals. The collected fractions are re-analyzed and quantified.

Conclusion 

The Agilent 1290 Infinity II Preparative Open-Bed Fraction Collector is a versatile instrument with enhanced fraction bed capacity and flexibility for diverse collection scenarios. Flow path and mechanics have been optimized for the highest purity and recovery of collected fractions. This Technical Overview demonstrates that fractions with high purity and recovery (above 98 % and 96 %, respectively) can easily be collected with the peak-based fraction mode triggered by UV signal only or a combination of the UV and MSD signals. When the peak-based, collecting time slices fraction mode is used to cut peaks into six or more slices, the purity of the pooled slices is on the same level as in peak-based collection. Sample loss caused by movement of the fraction valve between the slices is only marginal, yielding recoveries of the pooled fractions above 92 %.