Liquid and gas chromatography have become staples in medical, proteomics, metabolomics and genomics applications.
Driven by rapid growth in forensics, biotechnology, disease diagnostics and environmental regulations, chromatography systems have become a laboratory staple. Used for the separation of complex mixtures, detection of illicit drugs and the production of pharmaceuticals, the biotechnology and pharmaceutical industries are the prime users of chromatography techniques. Globally, as these industries are expanding, the demand for these techniques and instrumentation is increasing. Growth in the use of chromatography techniques in research activities in the fields of medicine, proteomics, metabolomics and genomics also propels the growth of the market.
According to Transparency Market Research data, the global chromatography market is expected to grow at a CAGR of 4.1% from $6.6 billion in 2011 to $8.9 billion in 2017. The global chromatography accessories and consumables market, according to a Research and Markets report, is an ever-changing technology-driven market—which includes columns, autosamplers, vials and detectors—and was valued at an estimated $2.6 billion in 2013, and is forecast to grow at a CAGR of 5.7%, to reach $3.5 billion by 2018.
Chromatography is categorized into three types comprised of liquid, gas and other chromatography techniques—which includes ion exchange and flash. While in most economies the chromatography market is shifting towards advanced chromatographic technologies, like ion exchange, liquid and gas chromatography will continue to sustain in the market. These techniques are still main staples in most forensics, life science and biotechnology laboratories. And this trend is highlighted through many products offered by Agilent Technologies, Phenomenex, Thermo Fisher Scientific, Shimadzu Scientific Instruments, Waters Corp. and more.
LC enhances life science separations
Liquid chromatography (LC) is a separation technique in which the mobile phase is a liquid. The technique can be conducted either in a column or a plane and generally uses small packing particles and relatively high pressure. Chromatographic separations of proteins are based on their difference either in size (size-exclusion chromatography), electric charges (ion-exchange chromatography) or hydrophobicity (reversed-phase chromatography).
JM Science offers a complete line of Shiseido HPLC CAPCELL PAK Columns with the Proteonavi HPLC column to separate proteins and peptides in reversed-phase mode. In reversed-phase mode, which uses a hydrophobic stationary phase, it’s known that proteins and peptides with higher-order structure are denatured in the course of retainment on the stationary phase after introduction to the column. Once the retention with denaturation occurs, molecules won’t migrate until the organic content of the mobile phase is raised to an appropriate level under a gradient program. Proteonavi utilizes high-purity silica with few metal impurities, and shows minimal irreversible adsorption for proteins and peptides. Its pore size is as wide as 30 nm, enabling large proteins enough interactions with the stationary phase.
The primary source of protein retention on reversed phase is hydrophobic interaction between hydrophobic parts of amino acid residues and alkyl chains in the stationary phase. When secondary interactions, such as coulombic interactions by acidic silanols (silica compounds) or metal impurities in the stationary phase, exist to a large extent, peak shapes of proteins will deteriorate due to their slow kinetics. Proteonavi allows better peak profiles and higher resolution among standard proteins in comparison with conventional wide-pore columns. This system is suitable for large-scale preparative separations.
For laboratories looking to produce greater amounts of information faster with chromatographic separations, Waters Corp. (Milford, Mass.) recently released the Waters CORTECS columns, a new family of 1.6-µm solid-core UltraPerformance LC (UPLC) columns. The column particles feature a solid, impermeable silica core encased in a porous silica outer layer where the interactions between the stationary phase and the analytes occur. Available in C18, C18+ or HILIC (hydrophilic interaction chromatography) chemistries, the column offerings include 30 configurations. The CORTECS C18 column is a general-purpose, high-efficiency, reversed-phase column offering balanced retention of acids, bases and neutrals at low and mid-range pH. The CORTECS C18+ column is a general-purpose reversed-phase column with a positively charged surface that delivers excellent peak shape for basic compounds at low pH. And the CORTECS HILIC column is designed for the retention of extremely polar analytes, while offering orthogonal selectivity versus C18 columns.
The growing number of scientists investigating glycans as disease markers and antibody-based biopharmaceuticals now have an effective new tool for the challenging separations of glycans and glycan isomers. Until now, glycan isomers couldn’t be separated by conventional high-performance liquid chromatography (HPLC), but Thermo Fisher Scientific, Waltham, Mass., has introduced the GlycanPac AXR-1 column, which helps users achieve greater resolution of glycans.
These columns are designed for high resolution of labeled and unlabeled glycans, and are compatible with fluorescence and mass spectrometry detection methods. Mass spectrometry (MS) has emerged as a powerful tool for determining the structures of glycans, and the GlycanPac AXR-1 column is designed to enable researchers to harness the power of high-resolution accurate-mass MS at high-throughput rates. The column is also designed to separate both labeled and native glycans based on the availability of samples. Native glycan separation allows researchers to eliminate the fluorescent labeling step and increase throughout without eroding performance.
Size-exclusion chromatography (SEC) is used for protein sizing and studying aggregates and bioconjugates, which are biomolecules that share a covalent link. Even at the lowest concentrations, these “misfolded” proteins that accumulate together in recombinant protein and monoclonal antibody biologics can be toxic and cause disease-producing effects. They must be identified and remediated at every phase of the drug development lifecycle. Agilent Technologies Inc., Santa Clara, Calif., has introduced the 1260 Infinity Multi-Detector Bio-SEC solution, a SEC system that features advanced light-scattering detection capabilities, fully bio-inert instrumentation, high-resolution columns and intuitive software. These elements enhance SEC for protein aggregate analysis.
The 1260 Infinity Multi-Detector Bio-SEC solution provides high sensitivity and accuracy through advanced detection capabilities and sophisticated yet intuitive software capabilities. All capillaries and fittings throughout the autosampler, column compartment and detectors are metal-free, so the biomolecules in a user’s sample come in contact only with ceramics or PEEK (polyether ether ketone). The dual-angle light-scattering detector measures the scattering of monochromatic laser light by polymer molecules at 15- and 90-degree angles. Combined with data from a concentration detector, light-scattering detection gives absolute molecular weights without the need for column calibration, accurate assessment of molecular size and radius of gyration and direct determination of long-chain branching. Used throughout the drug development lifecycle, the technology can simplify and speed up workflow, reducing the time and expense associated with bringing biopharmaceuticals to market.
GC for the masses
Commonly used in analytical chemistry for separating and analyzing compounds that can be vaporized without decomposition, gas chromatography (GC) is typically applied to the purity testing of substances or separating the different components of a mixture, especially within the life science arena. GC can also help identify a compound; and in preparative chromatography, GC can be used to prepare pure compounds from a mixture. In this method, the mobile phase is a carrier gas (helium or nitrogen) and the stationary phase is a microscopic layer of liquid on an inert solid support inside a column. Physical components of a GC system include autosamplers, inlets, detectors and columns.
A new solution for the identification of unknowns in metabolomics and small-molecule research, Bruker announced its GC-APC II source at Pittcon 2014, which allows routine and convenient combination of complementary techniques, which include GC, MS and atmospheric-pressure chemical ionization. In contrast to rigid conventional GC-APCI-MS interfaces, it can be easily mounted and dismounted within minutes, with no need for tools. The flexible transfer line means that an exact alignment of GC and MS is no longer required, providing more freedom in positioning the GC system, and making better use of laboratory space.
Efficient control of the heating of the transfer line preserves the GC separation of Bruker’s 436-GC and 456-GC. The redesigned ionization chamber provides high sensitivity and low background noise in combination with well-resolved GC peaks, delivering improved dynamic range and up to tenfold gains in sensitivity. The interface can be combined with any Bruker electrospray ionization (ESI) mass spectrometer and its unique calibrant reservoir enables software-controlled calibrant delivery. The calibrant can be added for data-file recalibration in each GC run.
Expanding the horizons of fast and transportable GC applications with innovations needed for accurate on-site analysis, INFICON, East Syracuse, N.Y., introduced its Micro GC Fusion at Pittcon 2014, the first temperature-programmable micro GC that offers fast temperature ramping using capillary columns. Built on the company’s proven micro GC technology, coupled with rapid temperature ramping, the Micro GC Fusion covers up to C12 gas phase analysis with excellent sensitivity and repeatability. The Micro GC system is equipped with a microelectromechanical system (MEMS) thermal conductivity detector (TCD) that offers a 1 ppm detection limit. The temperature-programmable GC column focuses late eluting peaks, providing advanced control over peak resolution and significant sensitivity gains for heavier hydrocarbons.
The Micro GC system is based on a modular GC design, where each module is comprised of an injector, a temperature-programmable capillary column and a detector. The instrument can be outfitted with one or two modules to meet most application needs. Users can easily exchange a GC module on-site in minutes to quickly adapt to new applications or conduct instrument maintenance. The modules can also be easily adaptable for OEM integration. The MEMS injector in the GC module eliminates the complex valve-switching programming that is typical in a benchtop GC, and the integrated sample conditioner in the instrument allows the user to accurately analyze sample gas streams at input pressures of up to 1,000 psi. The system runs on browser-based software, is equipped with a multi-touch front panel display and wireless network protocol and has an interface that can adapt to any display dimension, allowing users to operate the micro GC from a tablet, laptop or desktop PC.