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Heat-assisted extraction for the determination of methylarginines in serum by CE.

ABSTRACT: Methylarginines (MAs) are potent vasoconstrictors that have been reported to be present at elevated concentrations in the blood of patients suffering from cardiovascular disease. To determine the diagnostic potential of MAs for cardiovascular disease, a method capable of rapidly quantifying their endogenous concentrations from serum samples is required. To that end, a heat-assisted extraction method was developed. Serum was first rapidly heated, causing it to congeal into a gel, and then subjected to solid-liquid extraction. The extraction solution was then derivatized with a fluorogenic dye and analyzed by CE-LIF to permit quantitation of the MAs. This heat-assisted extraction procedure allowed a no-net-dilution extraction of the analytes to be performed into a solvent compatible with the subsequent CE analysis. This enabled direct detection of low-abundance analytes, such as MAs, without the need for a preconcentration step. This sample preparation method was compared with a commonly used solid-phase extraction method for MA analysis. Endogenous MA concentrations determined by both the heating and solid-phase extraction methods were found to be in good agreement with each other and with values previously reported in the literature.

SUBMITTER: Linz TH

PROVIDER: S-EPMC3744101 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Heat-assisted extraction for the determination of methylarginines in serum by CE.

Linz Thomas H TH Lunte Susan M SM

Electrophoresis 20130601 11

Methylarginines (MAs) are potent vasoconstrictors that have been reported to be present at elevated concentrations in the blood of patients suffering from cardiovascular disease. To determine the diagnostic potential of MAs for cardiovascular disease, a method capable of rapidly quantifying their endogenous concentrations from serum samples is required. To that end, a heat-assisted extraction method was developed. Serum was first rapidly heated, causing it to congeal into a gel, and then subject ...[more]

PMID: 23417924

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Project description:Polyphenolic compounds play an essential role in plant growth, reproduction, and defense mechanisms against pathogens and environmental stresses. Extracting these compounds is the initial step in assessing phytochemical changes, where the choice of extraction method significantly influences the extracted analytes. However, due to environmental factors, analyzing numerous samples is necessary for statistically significant results, often leading to the use of harmful organic solvents for extraction. Therefore, in this study, a novel DES-based shaking-assisted extraction procedure for the separation of polyphenolic compounds from plant samples followed by LC-ESI-QTOF-MS analysis was developed. The DES was prepared from choline chloride (ChCl) as the hydrogen bond acceptor (HBA) and fructose (Fru) as the hydrogen bond donor (HBD) at various molar ratios with the addition of 30% water to reduce viscosity. Several experimental variables affecting extraction efficiency were studied and optimized using one-variable-at-a-time (OVAT) and confirmed by response surface design (RS). Nearly the same experimental conditions were obtained using both optimization methods and were set as follows: 30 mg of sample, 300 mg of ChCl:Fru 1:2 DES containing 30% w/w of water, 500 rpm shaking speed, 30 min extraction time, 10°C extraction temperature. The results were compared with those obtained using conventional solvents, such as ethanol, methanol and water, whereby the DES-based shaking-assisted extraction method showed a higher efficiency than the classical procedures. The greenness of the developed method was compared with the greenness of existing procedures for the extraction of polyphenolic substances from solid plant samples using the complementary green analytical procedure index (ComplexGAPI) approach, while the results for the developed method were better or comparable to the existing ones. In addition, the practicability of the developed procedure was evaluated by application of the blue applicability grade index (BAGI) metric. The developed procedure was applied to the determination of spruce root samples with satisfactory results and has the potential for use in the analysis of similar plant samples.

Dataset Information

Heat-assisted extraction for the determination of methylarginines in serum by CE.

Publications

Heat-assisted extraction for the determination of methylarginines in serum by CE.

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