Monolithic silica columns for high-efficiency chromatographic separations

Tanaka, Nobuo; Kobayashi, Hiroshi; Ishizuka, N.; Minakuchi, Hiroyoshi; Nakanishi, Koji; Hosoya, Ken; Ikegami, Tohru

doi:10.1016/s0021-9673(01)01582-5

Cited by 485 publications

(304 citation statements)

References 42 publications

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“…Another property of these conventional columns is their pressure limit. The pressure drop increases with smaller particle size which, however, reduces eddy diffusion and results in greater column efficiency [13][14][15]. In recent years, monolithic silica gel columns are used increasingly more often for the separation of lipids, and several applications have been published [16,17].…”

Section: Introductionmentioning

confidence: 99%

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

Graeve

Janssen

2009

Journal of Chromatography B

124

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a b s t r a c tAn improved HPLC method is presented, which allows separation and quantification of a broad range of lipid classes of marine zooplankton with special regard to neutral lipids. Marine zooplankton species often produce high amounts of exceptional lipids, especially at high latitudes, in order to cope with the harsh environmental conditions and strong seasonality in food supply. Major neutral lipid classes are wax esters, triacylglycerols, diacylglycerol ethers, free fatty alcohols and sterols. Neutral and polar lipids were separated and identified on a monolithic silica column (Chromolith ® Performance-Si) using high performance liquid chromatography (HPLC) with an evaporative light scattering detector (ELSD). The method resolves a broad spectrum of lipids, varying in polarity from squalene to lysophosphatidylcholine in a single run. The total run time was 35 min including column re-equilibration. The calibration was made at levels of 0.1-60 g lipid/injection, but a 10-15-fold greater amount can be injected if single lipid classes need to be separated, e.g. for further determination of individual fatty acids. The method was applied to representative Arctic zooplankton species (copepods, pteropods, euphausiids and ctenophores) that are known to biosynthesize in particular neutral lipids like diacylglycerol ethers and free fatty alcohols.

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Section: Introductionmentioning

confidence: 99%

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

Graeve

Janssen

2009

Journal of Chromatography B

124

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“…In order to achieve a complete sample-in-answer-out system, it is imperative to integrate sample preparation and detection modalities. While DNA capture via solid phase supports, such as ones created by packing microfluidic channels with silica beads or embedding the beads in sol-gel matrices 10,11 , is more conducive to integration into miniaturized nucleic acid interrogation platforms, these devices still suffer from mechanical instability, including matrix shrinkage compromising DNA extraction efficiency 12,13 . In order to mitigate the mechanical issues and low reproducibility while maintaining a high surface area-to-volume ratio for efficient DNA capture, top-down photolithographic approaches were used to incorporate pillars in microfluidic channels 14 with various materials, including silica 15 , poly(methyl methacrylate) 16 , and polycarbonate 17 .…”

Section: Introductionmentioning

confidence: 99%

Sequence-Specific Electrical Purification of Nucleic Acids with Nanoporous Gold Electrodes

et al. 2016

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Nucleic acid-based biosensors have enabled rapid and sensitive detection of pathogenic targets; however, these devices often require purified nucleic acids for analysis since the constituents of complex biological fluids adversely affect sensor performance. This purification step is typically performed outside the device, thereby increasing sample-to-answer time and introducing contaminants. We report a novel approach using a multifunctional matrix, nanoporous gold (npAu), which enables both detection of specific target sequences in a complex biological sample and their subsequent purification. The np-Au electrodes modified with 26-mer DNA probes (via thiolgold chemistry) enabled sensitive detection and capture of complementary DNA targets in the presence of complex media (fetal bovine serum) and other interfering DNA fragments in the range 50 to 1500 base pairs. Upon capture, the non-complementary DNA fragments and serum constituents of varying sizes were washed away. Finally, the surface-bound DNA-DNA hybrids were released by electrochemically cleaving the thiol-gold linkage and the hybrids were iontophoretically eluted from the nanoporous matrix. The optical and electrophoretic characterization of the analytes before and after the detection-purification process revealed that low target DNA concentrations (80 pg/µL) can be successfully detected in complex biological fluids and subsequently released to yield pure hybrids free of polydisperse digested DNA fragments and serum biomolecules. Taken together, this multifunctional platform is expected to enable seamless integration of detection and purification of nucleic acid biomarkers of pathogens and diseases in miniaturized diagnostic devices.

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“…Wen et al [14] developed a tetramethyl orthosilicate-grafted monolithic solid-phase in a capillary-based extraction system, which was further modified for microchip-based extraction [15]. The monolithic column was characteristic of porous structure [16,17], allowing low pressure and high surface area.…”

Section: Introductionmentioning

confidence: 99%

Extraction of genomic DNA using a new amino silica monolithic column

Liu

Yang

et al. 2009

J of Separation Science

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Extraction of genomic DNA using a new amino silica monolithic column A new amino silica monolithic column was developed for DNA extraction in a miniaturized format. The monolithic column was prepared in situ by polymerization of tetraethoxysilane (TEOS) and N-(b-aminoethyl)-c-aminopropylmethyldimethoxysilane (AEAPMDMS). DNA was loaded in 50 mM tris(hydroxylmethyl)aminomethane -EDTA buffer at pH 7.0 and eluted with 300 mM potassium phosphate solution at pH 10.0. Under optimal condition, a 6.0-cm monolithic column provided a capacity of 56 ng DNA with an extraction efficiency of 71 l 5.2% (X l RSD). When the amino silica monolithic column was applied to extract genomic DNA from the whole blood of crucian carp, an extraction efficiency of 52 l 5.6% (X l RSD) was obtained by three extractions. Since the chaotropic-based sample loading and organic solvent wash steps were avoided in this procedure, the purified DNA was suitable for downstream processes such as PCR. This amino silica monolithic column was demonstrated to allow rapid and efficient DNA purification in microscale.

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Monolithic silica columns for high-efficiency chromatographic separations

Cited by 485 publications

References 42 publications

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

Improved separation and quantification of neutral and polar lipid classes by HPLC–ELSD using a monolithic silica phase: Application to exceptional marine lipids

Sequence-Specific Electrical Purification of Nucleic Acids with Nanoporous Gold Electrodes

Extraction of genomic DNA using a new amino silica monolithic column

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