Multiple heart-cutting 2D-LC closes the gap between comprehensive and heart-cutting 2D-LC
By Jens Trafkowski Agilent Product Manager, Liquid Phase Separations Business
LC samples that contain complex mixtures of components present a separation challenge, and sometimes you cannot resolve all important peaks with a single LC column. Two dimensional liquid chromatography (2D-LC) has become more popular in the analytical world, especially as commercially available 2D-LC systems have entered the UHPLC market. While heart-cutting 2D-LC is used in many laboratories today, the tasking technique of comprehensive 2D-LC is on the rise in those laboratories that must increase their chromatographic performance.
Figure 1. Standard heart-cutting 2D-LC analysis separates in the second dimension only the peaks enclosed in green boxes, while the Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting separates those in both red and green boxes.
Comprehensive 2D-LC is ideal for the most complex samples, but it incorporates many ultrafast gradients in the second dimension and therefore creates large amounts of data. Heart-cutting 2D-LC is used for isolated separation problems, but data points are lost when peaks elute from the first dimension while the second dimension gradient is running. As illustrated in the example in Figure 1, only the green-marked heart-cuts are analyzed in the second dimension; the red ones are lost and require two additional sample runs for analysis in the second dimension.
Get all the separation power you need, without acquiring unnecessary data
The new Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting solves this problem. It offers a smart valve setup to store peaks from the first dimension and analyze them as soon as the second dimension is free for the next analysis. Now all heart-cuts are analyzed in the second dimension – even those marked red in Figure 1.
Separate critical peaks for more accurate measurements
Chromatographic analyses of synthesis products – for example, pharmaceutical formulations or products from the chemical industry – frequently suffer from incomplete separation of the main compounds from their impurities, or incomplete separation of structurally similar impurities. To better understand the synthesis process and to ensure highest product quality, a common goal is to qualify precisely these side products. The Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting uses multiple 2D gradients to ensure the best separation at the critical parts of the chromatogram.
Figure 2 illustrates the technical setup of the valves for the Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting. The two sample loops of the 2D-LC valve are replaced with two 6-position/14-port valves. These 6-column selection valves are equipped with 12 loops in total to store heart-cuts from the first dimension, while the second dimension is running. An online monitoring function shows the actual filling and analyzing status of the system, giving all the necessary information about the valve cluster.
Figure 4. Smartest data analysis with the new Heart-Cut Viewer.
Develop complex methods more easily
To support the easiest method setup, the established easy-to-use 2D-LC software has been enhanced to enable the most straightforward setup of multiple heart-cutting methods. At a glance, you see all information about your 2D-LC method. Even better, the upload of a reference chromatogram during the method setup enables you to predict the peak-based or time-based heart-cuts for your sample analysis (Figure 3).
New tool speeds data analysis
Multiple heart-cutting chromatograms can be very complex, especially if you have taken many heart-cuts and therefore have performed numerous 2D-LC analyses. To analyze all the chromatograms generated in a multiple heart-cut analysis, we developed the new Heart-Cut Viewer. As shown in Figure 4, it captures and links together on a single screen:
The complete 1D chromatogram
A table with all heart-cuts, listed by their 1D retention time
The complete 2D chromatogram
Selectable single 2D chromatograms for more detailed analysis
Multiple heart-cutting closes the gap between conventional heart-cutting and comprehensive 2D-LC. You no longer have to perform several runs to analyze multiple peaks of interest in samples of medium complexity.
The new Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting delivers a complete ready-to-go solution with all necessary hardware. It enables the easiest method setup for multiple heart-cutting 2D-LC and unmatched data analysis with the new Heart-Cut Viewer. With this solution, 2D-LC becomes even easier-to-use, more high-performing, and more applicable for every analytical chemist. For additional information, watch the video about the new Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting.
Standard heart-cutting 2D-LC analysis separates in the second dimension only the peaks enclosed in green boxes, while the Agilent 1290 Infinity 2D-LC Solution with multiple heart-cutting separates those in both red and green boxes.
Multiple heart-cutting valve setup (left) and online monitoring view (right).
Software enables visual cues for easy method setup of multiple heart-cutting.
Smartest data analysis with the new Heart-Cut Viewer.