ACQUITY UPLC H-Class Bio

Multi-solvent blending
The ACQUITY UPLC H-Class Bio System provides laboratories with the ultimate in flexibility with its multi-solvent blending capability, allowing for binary, ternary, or quaternary gradient operation. The addition of the optional, six-port solvent select valve greatly expands your solvent choices allowing you to optimize method conditions.

Precise sampling that’s reliably quantitative
The system’s flow-through-needle sample manager is accurate and precise with excellent sample recovery and no carryover, resulting in more reliable and reproducible results.

Accurate column temperature control
A variety of column heaters and multi-column managers (with column switching) offer the lowest possible dispersion and precise temperature management for the highest retention time precision, ensuring that you produce accurate results using methods that can be seamlessly transferred to other laboratories – down the hall or around the world.

Detection optimized for UPLC separations
Any UPLC separation is not complete without a full line of innovative, ultra-low-dispersion detectors designed to maintain peak integrity. Waters offers the widest range of UPLC-optimized detectors – from optical to mass spectrometry – enabling you to meet multiple detection requirements for biological applications.

The ACQUITY UPLC H-Class Bio System is uniquely designed for size-exclusion chromatography (SEC) and the characterization of proteins and their aggregates.

The ACQUITY UPLC H-Class Bio, coupled with AutoBlend Plus Technology, enables automatic calculations and proportioning of buffer stocks, based on pH or ionic strength, making any possible buffer combination available - maximizing productivity by eliminating the labor intensive step of manually mixing buffers.

In addition, Waters ACQUITY UPLC BEH200, 1.7-µm SEC column chemistry, utilizes the first commercial sub-2-µm particle giving biopharmaceutical laboratories the ability to separate and quantify monoclonal antibodies (mAb) and their aggregates in less than four minutes, faster than with conventional liquid chromatography, and still meet FDA requirements. Monitor mAb aggregation at speeds up to 10 times faster at comparable or better resolution than before, allowing you to overcome separation challenges and develop novel biotherapeutics in less time and at lower costs.

The ACQUITY UPLC H-Class features a quaternary gradient pumping system that can continuously blend four solvents for use in generating a chromatographic separation. Auto•Blend technology uses this capability to automate the formulation of mobile phases from reservoirs of pure solvents or concentrated stock solutions.

Auto•Blend technology can be applied to any combination of solvents, modifiers, buffers, and salts. Any sequence of isocratic, binary, ternary, or quaternary gradients can be utilized. We can then draw any percentage we want from any of those bottles to adjust the separation or actually create on-demand the mobile phase that we require for the separation.

Since mobile phase combinations and gradients are automatically blended on-demand to meet the optimum retention and selectivity of the separation, this technology is useful for routine assays, in complicated separations requiring multiple solvents, and in automatic method development or system flushing.

Auto•Blend Plus technology further extends this capability by automatically managing pH and ionic strength requirements for the mobile phase. The software calculates the proportions of buffer stocks required for desired conditions. Computation can be based on known pK values or on an empirical calibration table, making any possible buffer combination available. Users can also create their own library of combinations.

For example, in the separation of a mixture of analgesics, the ACQUITY UPLC H-Class has proven very suitable in providing a high-quality chromatogram. What we've also found is that we can use this quaternary system as a way to make our work easier and also to eliminate some of the sources of error.

In a binary separation, we run a gradient from a high percentage of water to a high percentage of organic solvent. We manually prepare an A solvent (water and formic acid) and a B solvent (acetonitrile and formic acid). We draw a portion of the flow from each of those two bottles and change that proportion as a function of time during the separation. We get very reproducible chromatography. But that chromatography depends on how well we mixed the solvents in those two bottles.

Alternatively, we can use the ACQUITY UPLC H-Class System in Auto•Blend mode. Here, instead of making two bottles of pre-formulated solvent, we use the A and the B solvent as pure water and pure acetonitrile. In the D line, we put a ten-fold concentrated solution of formic acid in water. We then draw from three of the four bottles to create the same chromatographic method.

When we take 88% of the flow from the A bottle and 10% of the flow from the D bottle, we still have the same percentage of formic acid in water at our initial conditions. As we change the proportions of water and acetonitrile during the gradient, the constant percentage flow from the D line ensures the required pH and salt concentration.

When we compare the ternary gradient with the binary gradient, we observe the same chromatography, exactly the same analytical results. So the ternary gradient does not compromise the quality of our analyses and gives us the opportunity to avoid errors and to put less effort into doing this routine work.

The advantage is that instead of having to measure out two pre-mixed solvent bottles, we only have to make the one measurement for the concentrated stock. It's easier to make one bottle than two, and because we're making fewer measurements, we have fewer opportunities for an error to creep into our measurements.

Auto•Blend and Auto●Blend Plus technologies make routine analyses easier to manage, easier to execute by reducing the amount of work to be done in preparing complex mobile phases, easier to transfer methods to other laboratories, and it makes the whole laboratory process more efficient by reducing the possibility for errors during the separation.

For the analysis of biomolecules including monoclonal antibodies, recombinant proteins, DNA/RNA, and vaccine components, the ACQUITY UPLC H-Class System and Protein-Pak Hi Res, Ion-Exchange (IEX) columns give biopharmaceutical manufacturers the ability to reproducibly characterize various charge states of intact biomolecules - with greater resolution and speed. This improved monitoring capability helps manufacturers ensure high product quality and efficacy.

Waters IEX columns were developed to assist in the UPLC characterization of recombinant proteins and monoclonal antibodies found in many of today's biopharmaceutical therapeutics. These non-porous, high-ligand density particles overcome the effects of protein diffusion on peak volume and resolution experienced when using traditional porous IEX particles for large molecule separations. In addition, the novel particle and surface chemistry delivers high sample loading capacities and component resolution while minimizing column fouling.

AutoBlend Plus Technology for the ACQUITY UPLC H-Class Bio System is a software technique that maximizes your separation selectivity using the four-solvent capability of the system's quaternary solvent manager. Desired mobile phase blends are automatically proportioned to match the requirements of the separation ensuring optimum retention and selectivity.

In reversed-phase separations, for example peptide maps or protein analysis, the method can be adjusted by changing the concentration of mobile phase modifier and the organic solvent. For these applications, a series of experiments with different concentrations of TFA with gradients of increasing acetonitrile or isopropanol are typically employed. Instead of manually preparing many solvent mixtures, the ACQUITY UPLC H-Class Bio System makes blends from pure solvents and concentrated stocks on demand.