purpose
In order to confirm that the ACQUITY UPLC® I-Class system can minimize the extra-column band extension, and thus the separation effect is better when performing high-resolution and high-throughput UPLC® separation. The importance of these improvements will be explained below through impurity analysis and ballistic gradients.
background
It has been confirmed that in various applications, the use of columns packed with sub 2-m particles can improve the peak capacity and resolution of chromatographic separation, thereby greatly improving the resolution and throughput.
However, to maximize the degree of separation possible for a given separation, it is necessary to minimize system diffusivity. Any liquid lines or connections that are part of the system flow path behind the injector can cause the extra-column band to broaden. Including injection valve, solvent preheating device, connecting pipeline, accessories, and optical flow cell. Many suppliers have tried to improve the diffusibility of UHPLC systems, but with little success. Although it can reduce the diffusivity, it still can not achieve the best to obtain all the advantages of narrow-pore UPLC column (inner diameter 2.1 mm). These columns require lower flow rates, which makes the analysis of each sample higher return on investment, so that efficient separation can be performed at sufficient resolution.
solution
The ACQUITY UPLC I-Class system can reduce the band distribution outside the column. The optical path of the newly designed UV detector flow cell is the same as that of the previous ACQUITY UPLC, and the same high sensitivity can be obtained. In addition, the fluid piping and connections have been redesigned to further reduce the band spread. A solvent preheater must be used to minimize temperature gradients that can cause dispersion effects on the column. Therefore, the volume of the solvent preheater should be small enough to ensure that the sample plug reaches the head of the column with minimal diffusion, and it can provide excellent solvent heating performance even at high temperatures and high flow rates. According to the needs of your laboratory, you can choose one of the two sample managers to construct the ACQUITYUPLC I-Class system. Regardless of whether a fixed-loop (SM-FL) or flow-through needle (SM-FTN) sampler is used, it has been caused by the sampler through the use of small volume needle ports, connecting tubing, and internal valve channels The diffusibility is minimized. In general, the design of the fixed-loop sampler allows the extra-column band to expand to a lesser extent, because it reduces the volume of the syringe flow path. By optimizing each component, the extra-column band extension has been significantly reduced compared to any other commercially available LC system. Table 1 summarizes the band extension values ​​obtained after using various systems, including UHPLC systems.
The ACQUITY UPLC family outperforms all other systems in maintaining the integrity of ultra-efficient separation, of which ACQUITY
The band extension of the UPLC H-Class system is reduced to 9 _L, while the ACQUITY UPLC I-Class system is reduced to as low as 5.5 _L.
The reduced system diffusivity can directly lead to increased resolution of the ACQUITY UPLCI-Class system. The separation can reach a ballistic gradient while maintaining the resolution in a typical analytical gradient. Figure 2 illustrates the results of impurity analysis of tetracaine.
The ACQUITY UPLC I-Class system and the UHPLC system purchased from supplier B were used for separation under the same conditions.
The resolution of ACQUITY UPLC I-Class is significantly better. Supplier B's system installed a flow cell with an optical path length of 60 mm according to its recommendations. It was found that it produced a significant band extension so that no shoulder peaks could be measured.
summary
The ACQUITY UPLC I-Class system has unmatched performance and can be used in today's most challenging separation tasks. Whether your laboratory needs to increase the resolution of the separation or increase the sample throughput, its flexible system configuration can minimize the extra-column band expansion on the UPLC chromatography column to obtain the best separation performance.
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