There are several types of packing materials for waters columns. These materials are classified according to particle size and column dimensions. In this chapter, we will discuss the packing materials for size exclusion chromatography (SEC) using organic solvents. The products of the Styragel line are suitable for general SEC applications using organic solvents, Ultrahydrogel is used for water-soluble polymers, and Protein-Pak is suitable for proteins. The Waters Corporation provides additional information about the packing materials used in its columns.
2.7 micron
The 2.7 micron water column is the most widely used chromatography media for the separation of small particles. This column shows the largest number of plates per square meter than the 1.6 and 2.0 mm particle sizes. Several other benefits of this column include its excellent packed bed homogeneity and high efficiency. Here are some of the most important advantages of this column. If you are interested in purchasing one, you will want to consider a few points before making your decision.
First, column stability is an important issue for users. Injection-induced pressure pulses require high pressures to produce the separation you desire. The kinetic plots presented in Figure 6 show the dead time of the column as a function of plate number N at different pressures. The maximum pressure recommended by the manufacturer was used for the analysis. Higher pressures allow longer columns with larger particles, but this will increase the separation time.
The sub-2 mm SPP particles show similar advantages and disadvantages as the sub-2 mm particles, but with less size restrictions. However, the disadvantages of the sub-2.7 mm SPP particles are not as pronounced as those of the sub-2 mm particles. The sub-2.2 mm SPP particles are also commercially available. The sub-micron particles also have less practical limitations and have a good efficiency to back pressure ratio.
C18+
Waters CORTECS Solid Core C18+ Columns are designed to provide superior performance, while maintaining the same peak capacities as other solid-core columns. They feature a positively charged surface modification to improve selectivity and resolution of basic compounds. The columns are available in 48 unique configurations and can be shipped worldwide. They can be configured to meet all your analytical needs and are ideal for applications that require high-quality results. For more information, please see the C18+ Waters Columns page.
The new CORTECS Solid-Core Particle Columns from Waters are designed to improve the performance of your HPLC or UHPLC system. They feature balanced retention of acids, bases, and neutrals, and are compatible with Waters ACQUITY UPLC systems. The C18+ Waters Column also features a positively charged surface that provides excellent peak shape for basic compounds even at low pH levels. Waters C18+ Columns provide the ultimate in LC/MS efficiency and versatility.
In terms of selectivity, C18+ Waters Columns are the most widely used HPLC column for protein separation. The Harmony C18 HPLC column has equivalent selectivity to Waters Symmetry C18HPLC columns. Its excellent reproducibility and column lifetime make it a popular choice for chromatographers who need the performance and reproducibility of a Symmetry column. It also comes in bulk quantities and C8 and C4 phases.
HILIC
Conventional HILIC waters columns have limitations for retention of highly polar and moderately polar analytes. In addition, HILIC is not suitable for untargeted metabolite profiling, as polar compounds elute easily with matrix interferences. Additionally, phosphorylated metabolites may never elute from the column because they adsorb to the metal surfaces of the column. To overcome these problems, chromatographers must condition the column overnight before running the sample.
The mobile phase used in HILIC separations usually consists of water-miscible polar organic solvents, and high percentages of volatile organic phase provide increased sensitivity in MS with ESI. Furthermore, the organic solvent-rich SPE eluate can be introduced directly onto the column without reconstitution or evaporation. Once this step is complete, the eluate is ready for chromatographic analysis.
HILIC columns are available in a variety of sizes and types. Waters Atlantis ™ Premier BEH column uses an ethylene-bridged hybrid chemistry with MaxPeak High Performance Surfaces (HPS) column hardware. This allows researchers to avoid complex and lengthy column conditioning protocols. HILIC columns are available in sub-two-micron, 2.5-micron, and five-micron particle sizes.
Large scale
In order to achieve higher throughput and increased data quality, Waters Corporation has introduced its ACQUITY(tm) PREMIER Columns. These columns are compatible with any brand of UHPLC system and are designed to improve sample throughput, assay-to-say reproducibility, and overall confidence in analytical results. For more information, please see the datasheets linked below. Here are some other features that these columns offer.
The Taylor column mixing rate is estimated from ship-derived transient tracer data and is approximately 1000 m2/s, the same rate that is associated with the ACC’s transient mesoscale eddies. Because the Taylor column acts as a steady blender, the waters near the WSC are retained for extended periods of time. Because of this, their composition can undergo considerable modification. However, such measurements can be challenging.
Fishing in the water column
While commercial fishing gear is extremely effective for catching fish, its methods can have detrimental effects on the ocean’s natural habitats. Heavy chains and dredges can destroy delicate corals and sponges, reducing the habitat for bottom-dwelling fish and shellfish. Also, fishing in the water column can entangle and kill nontargeted species. Fortunately, fishing regulations can have the same conservation effect as full protection under the Antiquities Act.
CTD probes
CTD is an acronym for Conductivity-Temperature-Distance. A CTD measures three important elements of water: temperature, conductivity, and depth. Temperature and conductivity of water are used to determine the salinity of the water column. The probe may be incorporated into a series of sampling bottles known as a rosette. Water sampling bottles are usually closed at pre-set depths to collect water samples. These samples are then analyzed for biological parameters.
Different types of CTDs are available. Profiling CTDs can be deployed from a ship or can be integrated with autonomous vehicles, gliders, and Argo floats. In both cases, a package can include several accessories. Niskin bottles, for example, are used to collect samples of water. Other accessories include an Acoustic Doppler Current Profiler, which measures the horizontal velocity of water, and oxygen sensors, which measure the dissolved oxygen content of water.
The properties of water change as the depth increases. At the surface, oxygen levels are high and fall rapidly. This decreases as the water descends. At the base of the photic zone, the amount of chlorophyll drops rapidly. As chlorophyll depends on photosynthesis to survive, it cannot perform photosynthesis below this depth. A CTDS can help scientists understand these changes in water and develop better tools to measure their data.
Acquity Column Manager
The ACQUITY UPLC(r) System removes all barriers to the performance of a column, allowing even smaller particles to reach their theoretical capacity. The ACQUITY UPLC system delivers mobile phase at pressures up to 1030 bar (15000 psi) without compromising column performance. Whether you need a water-based chromatography solution or a column for a biopharmaceutical application, the ACQUITY UPLC column family delivers the solution.
With its flexible features, the ACQUITY UPLC Column Manager is ideal for high-throughput UPLC sample processing. The ACQUITY CM-30S supports up to four columns of different chemistries, and it comes standardized with low-volume active solvent pre-heaters to ensure consistent column performance from run to run. The ACQUITY UPLC Column Manager supports two or four columns of either 150 or 50 mm in length.
The Waters ACQUITY 30cm Single Zone Column Manager provides greater flexibility than a traditional column system, allowing for easy removal and replacement. It can accommodate eight separate columns and screen the chemistries and modifier strengths. This allows for quick analysis and a time-efficient scale-up to preparative chromatography. For more detailed information, read our complete review of the Acquity Column Manager for Waters.