As new products introduced this year have shown, HPLC systems continue to evolve. Speed, resolution and ease of use remain key criteria for end-users so as to increase productivity. Vendors continually optimize and develop HPLC techniques, such as UHPLC and LCxLC, to meet these demands. Agilent Technologies, Shimadzu and Thermo Fisher Scientific each launched new HPLC products this year. The products and the company’s positioning of them reflect technical and application trends in the HPLC and UHPLC market, as well as how companies intend to drive market growth. Companies are adding to their product lines, leveraging developments in software and columns, and adapting to the maturation of the UHPLC market. The total analytical HPLC market is estimated to top $4 billion in revenues this year (see table, page 6).

A major UHPLC product introduction this year was the Thermo Scientific Vanquish UHPLC Platform from Thermo Fisher Scientific. The product line adds a new tier to the company’s UHPLC offerings, according to Fraser McLeod, vice president and general manager of HPLC for Thermo Scientific. Thermo Scientific’s UHPLC product lines consist of the Ultimate 3000 platform, which was acquired through the purchase of Dionex (see IBO 12/15/10). “It adds a tier to the product that cannot be addressed by the Ultimate 3000 Series,” said Mr. McLeod.

The Vanquish operates at a pressure of up to 1,500 bar (22,000 psi). The company’s highest-pressure system prior to the Vanquish was the Accela, which operated at 1,350 bar and was discontinued in December 2012. As Mr. McLeod told IBO, the Vanquish’s higher pressure range enables greater productivity by increasing the speed and resolution of UHPLC analyses. “There is always a need, especially in R&D—because it was designed predominantly for the R&D user base—to do more than you can currently do; to be able to do more precise experiments,” he said. “So to be able to achieve more resolution than you currently can, to see more of what’s in the substance that you are trying to analyze, to have higher throughput so you can do more rapid experiments, to do more iterations of experiments, and then get quicker answers from the outcome of those experiments by providing higher throughput and higher resolution.”

System design and components are key aspects of taking advantage of the higher speed and greater resolving power of UHPLC. The Vanquish features a new pump design for greater precision, and thus more consistent results, and a new injection valve for improved reliability, according to Mr. McLeod. The Charger Module is designed to accommodate 8,832 samples so as to leverage UHPLC’s speed. Throughput was an important consideration, according to Mr. McLeod, who said it addresses the issue of how to load more samples to take advantage of UHPLC’s speed. “If I am running 10 times as many samples, where am I going to put them. Most autosamplers are specified to the number of samples you could run under an HPLC.”

To maximize productivity gains, the Vanquish is designed to work with the company’s new line of Accucore Vanquish columns, superficially porous particle (SPP) columns with 1.5 µm particles, available in C18 reversed-phase chemistry. It is Thermo Scientific’s first sub-2µm SPP column. The new columns used with a 1,500 bar instrument means that “you can take full advantage of the extra resolution potential in that column. You can take advantage of the extra speed potential within that column,” stated Mr. McLeod.

By providing this increased resolution and throughput, the Vanquish can meet a different set of customer needs compared to the Ultimate 3000, according to Mr. McLeod. “What became clear in the feedback from our customers in the last few years is that, although it meets many of their needs, for certain workflows and certain applications, there were certain needs that it didn’t quite get to.”

Introduced in July, the Vanquish is used in several end-markets. “It has clearly already shown success in biopharma R&D workflows, which requires either high resolution or high throughput. It has already shown good traction in the chemical market, predominantly for apps looking for higher throughput. And it’s shown good traction in food-science laboratories, either again for high resolution or for higher throughput.”

The system also sought to counter end-users’ perception of UHPLC as being more difficult than HPLC. “[We wanted to] make the user experience as straightforward as possible,” stated Mr. McLeod. As a result, the system utilizes the Viper fittings and is designed for easier internal access. To simplify method transfer, the heating compartment operates in either isothermal or adiabatic modes.

Optimizing the use of UHPLC by making it flexible, increasing user friendliness, and adding to performance is also evident in Agilent Technologies’ latest UHPLC system. In November, the company began shipments of the 1290 Infinity II system, offering it for the same price range as the classic 1290 Infinity. The classic 1290 Infinity, whose enhanced version launched in 2010, is still available. The 1290 Infinity II operates at 1,300 bar and is the premium system in Agilent’s Infinity platform. The 1290 Infinity II is a second-generation UHPLC system, according to Helmut Schulenberg-Schell, PhD, director of Business Development—Liquid Phase Separations, at Agilent. “The second generation is aimed at maximizing efficiency through the entire workflow—for chromatography experts, routine operators and lab management,” he said.

The statement illustrates the range of end-users that Agilent is addressing and the focus on efficiency. The flexibility is also evident through the number of modes and columns with which the system is designed to operate. “The 1290 Infinity II system offers one of the greatest automation possibilities to switch between different columns and solvents, between standard HPLC and UHPLC, and even between 2D-LC either multi-heartcutting or fully comprehensive 2D-LC,” said Dr. Schulenberg-Schell. The 2D-LC switch is new to the system. “This can all be realized with an accordingly configured 1290 Infinity II LC system and only by changing software parameters.”

New modules are designed to provide flexibility, improved operation and to further differentiate the system. The 1290 Infinity II features eight new instrument modules compared to the enhanced 1290 Infinity, said Dr. Schulenberg-Schell, including the High Speed Pump and the Flexible Pump, the Multisampler autosampler, the Multi-Column Thermostat, a diode array detector, and evaporative light-scattering detector. “What distinguishes the system is the 1290 Flexible Pump to address a wide application range, sample capacity per bench space, advanced sampler capabilities and backward method compatibility,” according to him.

Flexibility and usability, and thus efficiency, are also enabled by the ease of transfer from the previous 1290 system. “[A]ll existing 1290 Infinity methods can still be run on the new system, and all new modules are also compatible with the classic 1290 Infinity LC system—even the 1260 Infinity LC stacks,” he told IBO.

System design and features also accommodates varied end-markets. “With the flexibility it offers, the 1290 Infinity II LC is the ideal tool for method development in research labs for life science as much as for fine chemicals,” stated Dr. Schulenberg-Schell. “With the 2D-LC extension, it serves perfectly the exploration of impurities and complex fingerprints in food and biopharm labs. With maximizing efficiency, it will ultimately make inroads into 24 x 7 routine operation for a wide range of applications.”

Transfer of HPLC methods was just one of the challenges of UHPLC that the Infinity 1290 II addresses, according to Dr. Schulenberg-Schell. In addition, he told IBO, the system also minimizes carryover and improves low resolution by minimizing dead volumes in fitting, flow cells and needle seats. It also provides muti-method and multi-user operation, according to him. This higher utilization can result in a reduced fixed cost per sample.

A major introduction from Shimadzu this year was the official release of the Nexera-e Comprehensive LCxLC system this summer. The Nexera platform was introduced in 2010 (see IBO 3/15/10). Discussing LCxLC, Simon Robinson, HPLC Product manager at Shimadzu, stated, “The amount of separation power we get will always eclipse a single column.” As he told IBO, “This technique gives you exponential separation power versus traditional techniques. The other area where it really shines is when you need two different separation techniques to effectively split a sample.”

Nexera-e is built using standard modules and components of the Nexera platform, but features new software. Thus, Nexera systems can be upgraded to the Nexera-e with new software. “The Nexera-e does have a dedicated plumbing design, and the plumbing is easy to do. A whole installation, if we’re just talking about the plumbing, is 30 minutes,” said Mr. Robinson. Such upgradeability distinguishes the system, according to him. “We were one of the few vendors out there that when we build a system; we don’t put out a package system per se normally. Normally, [for] every one of our systems, the modules are put together to do whatever the customer’s requirement or specialty is.”

Describing potential end-users, Mr. Robinson said, “Typically, when I go into the lab, there would be one user who wanted to do very complicated things with the LC, and that was a two-dimensional person.” The range of options with LCxLC is complicated by column selection. “It’s critical. And there are inherent limitations of the column dimensions, flow rates and mobile phases you can use. It can be quite complex selecting the right combination. . . . It means it’s hard to do but it means you’ve also got a lot of variation of selection that could potentially provide some very novel and new results.”

The ChromeSquare data analysis and visualization software was developed by Shimadzu with Italian firm Chromaleont. Only recently have the software developments for GCxGC transitioned to LCxLC. “The reason being was that prior, the equipment really wasn’t reliable enough or consistent enough to the level where the results were worthwhile. The systems’ valving and timing and the software now is reliable enough to get you to the spot where the data you get is reproducible and reliable,” explained Mr. Robinson.

The software features a simple interface and all data are displayed on a single screen. In fact, Mr. Robinson expects further improvement in LCxLC to come through software development. “So it’s not like we would have to develop the hardware down a niche path to make it more akin to this technique. It relies on pump stability and valving. I believe what is going to start to mature is the software. I think it will mature further.”

Although the technique has found applications, especially in natural-products analysis, food science, chemicals and lipids, it has not yet found a “killer application,” according to Mr. Robinson. “Nobody has come up with one [application] where ‘this is the only way to do it’ and ‘it’s an analysis that has to be done.’” One area that is especially promising is food analysis. “I could see it potentially becoming routine in the food industry, just as they’ve got some very big challenges,” he said.