Miniaturized Columns for the Routine HPLC Lab: High Speed and Minibore Performance

Vonk, N.; Verstraeten, Will; Marinissen, J. W.

doi:10.1093/chromsci/30.8.296

Cited by 10 publications

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“…Using these data, an LOD of 0.045 nM can be estimated for natural samples, similar to that of pure solutions. These LOD, obtained with a 4.6 mm ID column, are comparable to those obtained by Pockington and co-workers (1990) using a 2.1 ID narrow-bore column (0.050 nM), which are known to provide 3 to 5 times greater sensitivity than conventional columns (Vonk et al 1992).…”

Section: Comparative Sensitivity and Reproducibility Of The Two Derivsupporting

confidence: 87%

Selective liquid chromatographic determination of trace domoic acid in seawater and phytoplankton: improvement using the o‐phthaldialdehyde/9‐fluorenylmethylchloroformate derivatization

Devez

Delmas

2013

Limnology & Ocean Methods

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Domoic acid (DA), the amnesic shellfish poisoning toxin (ASP), is an excitatory amino acid that can accumulate, under certain environmental conditions, in shellfish, finfish, birds, and mammals, by direct filtration or by feeding on contaminated organisms (Mos 2001). This potent neurotoxin with three carboxylic groups, responsible for its high polarity and hydrophilicity, is a cyclic amino acid and a secondary amine with a molecular weight of 311 Daltons. DA belongs to a neurotransmitter class of compounds with a structure very similar to an important neurotransmitter excitatory, acid glutamate, and indeed mimics glutamate in its interaction with some of its receptor subtypes. DA can damage the neurons by activating R-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and kainate receptors, causing an influx of calcium, and then be the cause of several symptoms such as nausea, disorientation, temporary amnesia and, in more serious cases, persistent memory loss and/or coma, and ultimately even death (Costa et al. 2010). This toxin was originally isolated from a red microalga Chondria armata by Japanese researchers (Takemoto and Daigo 1958) and is now reported to be produced in several species of marine diatoms from the genus Pseudo-nitzschia. DA was identified as the causative agent of the well-known tragic intoxication event observed in 1987 (Prince Edward Island from Cardigan Bay in Eastern Canada). More than one hundred people became ill and three victims died, by septic shock or pneumonia, 11-24 days after intoxication. Whereas most victims recovered within 10 days, after a period of confusion and nausea, the others continued to show signs of selective short-term memory loss (Bates et al. 1989). Additional consequences of the DA presence included the temporary closure of shellfish aquaculture industries and a strong impact on tourism activities (Pistocchi et al. 2012;Trainer et al. 2012).The detection limit of the method generally considered as the reference for particulate DA quantification in marine organisms (high-performance liquid chromatography with ultra-violet detection at λ = 242 nm, HPLC-UV) ranges between 13 and 250 nM depending on the detector (Quilliam 2003). This sensitivity threshold is often insufficient to follow the dynamics of both dissolved and particulate domoic acid AbstractDomoic acid (DA), a toxin produced worldwide by some species of the genus Pseudo-nitzschia, is responsible for contamination of marine molluscs, mammals, birds, and for human intoxication, and when detected in high levels results in closures of shellfish farms, thus causing severe economic losses to aquaculture. Studies on algal production of DA in cultures and field samples require sensitive methods capable of measuring trace concentrations of domoic acid. Measuring domoic acid concentrations at trace levels is still a significant challenge. A sensitive and reliable double derivatization using o-phthaldialdehyde-mercaptoethanol/9-fluorenylmethylchloroformate (OPA-MeSH/FMOC-Cl) followed by high-performance liq...

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Section: Comparative Sensitivity and Reproducibility Of The Two Derivsupporting

confidence: 87%

Selective liquid chromatographic determination of trace domoic acid in seawater and phytoplankton: improvement using the o‐phthaldialdehyde/9‐fluorenylmethylchloroformate derivatization

Devez

Delmas

2013

Limnology & Ocean Methods

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“…One topic of continuing interest in analytical chemistry is the development of smaller and faster methods of analysis. Examples in the field of separations include the production of microfabricated devices, as well as ongoing work in microbore HPLC and capillary electrochromatography. − These last two areas generally use a decrease in column diameter with an accompanying increase in column length to produce more efficient chromatographic systems for the analysis of complex samples. However, another way in which column size can be decreased is through a reduction in length.…”

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confidence: 99%

Development of Sandwich HPLC Microcolumns for Analyte Adsorption on the Millisecond Time Scale

Clarke

Hage

2001

Anal. Chem.

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A new class of columns is reported that uses only microgram quantities of active support and that provides for the retention of biological compounds and other analytes on the millisecond time scale. This was accomplished by packing standard HPLC supports into layers as small as 60 microm in length and using only 90 microg of support material. This provided columns with effective residence times in the millisecond time range when routine HPLC flow rates and pressures were used. The retention of analytes by such columns was examined under both adsorption- and diffusion-limited conditions. The RPLC adsorption of hemoglobin (a system with diffusion-limited retention) was found to give 95% binding in as little as 4 ms. The adsorption of fluorescein by an anti-fluorescein antibody column (an adsorption-limited system) gave 95% retention in 100-120 ms. One application examined for these columns was their use in a chromatographic-based competitive binding immunoassay. This used bovine serum albumin (BSA) as the model analyte, and fluorescein-labeled BSA was used for detection. The resulting approach had a contact time of 180 ms between the sample and an anti-BSA immunoaffinity microcolumn and provided a signal within 5-25 s after sample injection. The columns developed in this work should also be useful in other situations that involve a small amount of a stationary phase or that require short column residence times.

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“…Assim como ocorreu 40 anos atrás, atualmente o alargamento de banda extracoluna ainda vem sendo considerado um dos maiores desafios da miniaturização da cromatografia líquida (LC) [3][4][5][6][7][8] , uma vez que possui grandes implicações no desenvolvimento de sistemas instrumentais de alta eficiência destinados à escala capilar.…”

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Cromatografia líquida capilar: 2. Alargamento das bandas por efeitos extracoluna

Coutinho¹,

Lanças

2011

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ResumoO alargamento da banda é um fator altamente indesejável em todas as técnicas cromatográficas, uma vez que diminui a eficiência da coluna (menor número de pratos, N) e deteriora a resolução (menor Rs). Este efeito é particularmente crítico na cromatografia líquida capilar (c-LC) devido aos pequenos volumes da instrumentação, conexões, colunas, etc., empregados na técnica. Como parte de uma série de artigos a respeito da c-LC, neste trabalho serão abordados os principais fatores que ocasionam o alargamento das bandas cromatográficas em c-LC, assim como a maneira de evitá-lo. Palavras-chaveCromatografia líquida capilar; alargamento de bandas; fatores extra coluna; micro LC; LC capilar. Capillary liquid chromatography: 2. Band broadening due to extracolumn effects AbstractBand broadening is one of the major factors to be avoided in chromatographic techniques, once it decreases the column efficiency (the number of plates, N) and deteriorates the Resolution (Rs). This effect is particularly critical in capillary liquid chromatography due to the very low volumes, instrumentation, connections and columns employed in this technique. As a part in a series of papers dealing with c-LC, this work describes the major factors responsible for band broadening in chromatography, as well as the ways to minimize it.

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Miniaturized Columns for the Routine HPLC Lab: High Speed and Minibore Performance

Cited by 10 publications

References 0 publications

Selective liquid chromatographic determination of trace domoic acid in seawater and phytoplankton: improvement using the o‐phthaldialdehyde/9‐fluorenylmethylchloroformate derivatization

Selective liquid chromatographic determination of trace domoic acid in seawater and phytoplankton: improvement using the o‐phthaldialdehyde/9‐fluorenylmethylchloroformate derivatization

Development of Sandwich HPLC Microcolumns for Analyte Adsorption on the Millisecond Time Scale

Cromatografia líquida capilar: 2. Alargamento das bandas por efeitos extracoluna

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