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Effect of decreasing column inner diameter and use of off-line two-dimensional chromatography on metabolite detection in complex mixtures.

ABSTRACT: Capillary liquid chromatography coupled with electrospray ionization to a quadrupole ion trap mass spectrometer was explored as a method for the analysis of polar anionic compounds in complex metabolome mixtures. A ternary mobile phase gradient, consisting of aqueous acidic, aqueous neutral and organic phases in combination with an aqueous compatible reversed-phase stationary phase allowed metabolites with a wide range of polarities to be resolved and detected. Detection limits in the full scan mode for glycolysis and tricarboxylic acid cycle intermediates were from 0.9 to 36fmol. Using this system, 111+/-9 (n=3) metabolites were detected in Escherichia coli lysate samples. Reducing column I.D. from 50 to 25microm increased the number of metabolites detected to 156+/-17 (n=3). The improvement in number of metabolites detected was attributed to an increase in separation efficiency, an increase in sensitivity, and a decrease in adduct formation. Implementation of a second separation mode, strong anion exchange, to fractionate the sample prior to capillary RPLC increased the number of metabolites detected to 244+/-21 (n=3). This improvement was attributed to the increased peak capacity which decreased co-elution of molecules enabling more sensitive detection by mass spectrometry. This system was also applied to islets of Langerhans where more significant improvements in metabolite detection were observed. In islets, 391+/-33 small molecules were detected using the two-dimensional separation. The results demonstrate that column miniaturization and use of two-dimensional separations can yield a significant improvement in the coverage of the metabolome.

SUBMITTER: Edwards JL

PROVIDER: S-EPMC2710303 | biostudies-literature | 2007 Nov

REPOSITORIES: biostudies-literature

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Effect of decreasing column inner diameter and use of off-line two-dimensional chromatography on metabolite detection in complex mixtures.

Edwards James L JL Edwards Rachel L RL Reid Kendra R KR Kennedy Robert T RT

Journal of chromatography. A 20071010 2

Capillary liquid chromatography coupled with electrospray ionization to a quadrupole ion trap mass spectrometer was explored as a method for the analysis of polar anionic compounds in complex metabolome mixtures. A ternary mobile phase gradient, consisting of aqueous acidic, aqueous neutral and organic phases in combination with an aqueous compatible reversed-phase stationary phase allowed metabolites with a wide range of polarities to be resolved and detected. Detection limits in the full scan ...[more]

PMID: 17961581

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Project description:Intravenous (IV) fluid administration is critical for all patients undergoing care in a hospital setting. In-patient hospital practice, surgeries, and emergency care require functional IVs for fluid replacement and medication administration. Proper placement of IVs is vital to providing medical services. The ease of placement of an IV catheter, however, depends not only on the size of the catheter but also on provider experience and patient demographics such as age, body mass index, hydration status, and medical comorbidities present challenges to successful IV placement. Smaller diameter IV placement can improve success and there are instances where multiple small diameter catheters are placed for patient care when larger bore access is unattainable. Smaller inner-diameter catheters for anesthesia have functional constraints. Ideally, there would be a smaller catheter for placement that could function as a larger catheter for patient care. One solution is the idea of functionally responsive catheters. Here, we evaluated tubular-shaped hydrogels as potential functional catheters that can increase in inner diameter through fluid swelling using cross-linked homopolymers of polyacrylamide, PAM (10-40% w/w), and their copolymers with 0-8% w/w Poly-(Ethylene Glycol)-Diacrylate, PEGDA. For the PAM gels, the water transport mechanism was shown to be concentration-dependent Fickian diffusion, with the less concentrated gels exhibiting increasingly anomalous modes. Increasing the PEGDA content in the network yielded an initial high rate of water uptake, characterized by Case II transport. The swelling kinetics depended strongly on the sample geometry and boundary conditions. Initially, in a submerged swelling, the annulus expands symmetrically in both outward and inward directions (it thickens), reducing the internal diameter by up to 70%. After 1 h, however, the inner diameter increases steadily so that at equilibrium, there is a net (>100%) increase in all the dimensions of the tube. The amount of linear swelling at equilibrium depended only on the polymer volume fraction as made, while the rate of inner diameter expansion depended on the hydrophilicity of the matrix and the kinetics of sorption. This study serves as proof of concept to identify key parameters for the successful design of hydrogel-based catheter devices with expanding inner-diameters for applications in medical care.

Dataset Information

Effect of decreasing column inner diameter and use of off-line two-dimensional chromatography on metabolite detection in complex mixtures.

Publications

Effect of decreasing column inner diameter and use of off-line two-dimensional chromatography on metabolite detection in complex mixtures.

Similar Datasets

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