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Biomedical subjects

M D Luque de Castro

Publications and source records attributed to M D Luque de Castro.

At least 37 records · Page 2Linked to original sources

Ultrasound-assisted extraction for the analysis of phenolic compounds in strawberries.

A semiautomatic method based on application of ultrasounds has been developed to leach and hydrolyse phenolic compounds, such as naringin, rutin, naringenin, ellagic acid, quercetin and kaempferol, from strawberries. Two grams of lyophilized sample was placed into a sample cell and 5 mL of acetone containing hydrochloric acid was added. The cell was immersed in a water bath and sonicated for 30 s (duty cycle 0.8 s, output amplitude 50% of the nominal amplitude of the converter, applied power 100 W and with the probe placed 2 cm from the top surface of the extraction cell) for three times: each time 5 mL extractant displaced the previous extract. When the extraction was completed, the combined extracts were evaporated for 10 min, diluted to 10 mL with water adjusted to pH 8, and transferred to a cleanup-preconcentration manifold; here the analytes were retained in two in-series minicolumns packed with HR-P sorbent and then eluted with 4 mL methanol, and injected for individual separation-quantitation into a chromatograph-photodiode array detector assembly. Optimisation of the extraction was carried out using samples spiked with 4 mg kg(-1) of each analyte. Calibration curves using the standard addition in red strawberries typically gave linear dynamic ranges of 4-40 mg L(-1) for all analytes, except for ellagic acid (40-400 mg L(-1)). The r(2) values exceeded 0.98 in all cases.

Fragaria↗

Use of superheated liquids for the extraction of non-volatile compounds from wood: liquid chromatography studies.

A study of the extraction of non-volatile compounds from oak wood using superheated water-ethanol mixtures ranging from 10 to 60% ethanol is reported. Identification and characterization of the extracted compounds have been made by high performance liquid chromatography. The extraction has been performed using the static mode by single or repetitive cycles. The variables affecting the extraction process have been studied and their optimum values established (extraction time: 50 min; pressure: 40 atm; extraction temperature: 180 degrees C). The study allows comparing the non-volatile polyphenol fractions obtained in this way with those present in commercial samples with fully agreement between them. In addition, the method makes possible manipulation of the extract composition by changing the working pressure, temperature and water-ethanol ratio.

Chromatography, High Pressure Liquid↗

Ultrasound-assisted Soxhlet extraction: an expeditive approach for solid sample treatment. Application to the extraction of total fat from oleaginous seeds.

Conventional Soxhlet extraction assisted in the cartridge by ultrasound has been developed and used to extract the total fat content from oleaginous seeds such as sunflower, rape and soybean seeds. The application of ultrasound to the sample cartridge enormously decreases the number of Soxhlet extraction cycles needed for quantitative extraction of the fat, thus reducing the extraction time at least to half the time needed by the conventional procedures. The results agree well with those obtained by conventional Soxhlet extraction and the ISO reference method, both in terms of efficiency and precision. The repeatability of the proposed approach, expressed as relative standard deviation, was 0.9%; the within-laboratory reproducibility was 1.3%. Qualitative analysis of the extracted fat showed that the application of ultrasound does not change the composition of the oil.

Fats↗

Dynamic ultrasound-assisted leaching of essential macro and micronutrient metal elements from animal feeds prior to flame atomic absorption spectrometry.

A method for routine analysis of essential metal elements in animal feeds using a dynamic and automated ultrasound-assisted extractor is proposed here. Owing to the different concentration of the essential metal elements in the samples, two macronutrients-namely, Ca and Mg-and three micronutrients or trace elements-namely, Fe, Cu and Zn-were selected as models for the optimisation and characterisation of the proposed method by using experimental design methodology. Moreover, the extraction kinetics and influence of the particle size on the extraction efficiency were studied. The detection and quantification limits ranged between 2.9 and 40 microg kg(-1) and between 7.9 and 95.5 microg kg(-1), respectively. The precision, expressed as repeatability relative standard deviation (RSD) and as within-laboratory RSD, ranged between 1.86 and 5.66% and between 5.06 and 6.15%, respectively. The proposed approach allows the extraction of these metal elements from animal feeds with extraction efficiencies similar to those provided by the AOAC Official Method 968.08, but with a drastic reduction in both the extraction time (18 min versus 4.5 h) and sample handling, and using smaller volumes of extractant (an acid aqueous solution).

Animal Feed↗

Pressurised liquid-liquid extraction. An approach to the removal of inorganic non-metal species from used industrial oils.

Modified pressurised hot water is used for the development of a high pressure liquid-liquid extraction method for the decontamination of used industrial oils from inorganic non-metal species (chlorine, fluorine and sulphur). The oils were subjected to dynamic extraction with water modified with 5% v/v HNO3 at 200 degrees C as extractant. Under these working conditions the analytes were transferred to the aqueous phase. Spontaneous separation of the two immiscible liquid phases (the used oil and extract) takes place in the collection flask after extraction. The treated and untreated oil samples were oxidised and the chloride, fluoride and sulphate thus formed were determined by ion-chromatography. The method was applied to four oil samples from different locations in Spain. A residence time of approximately =10 min provided oil samples from which 88.3%, 89.4% and 89.4% of chloride, fluoride and sulphate, respectively, have been removed with respect to the initial concentration of each analyte in the oil. The repeatability, expressed as relative standard deviation (RSD), was of 11.9%, 13.7% and 7.2% for Cl(-), F(-) and SO4(2-), respectively; whilst the within-laboratory reproducibility yielded RSDs of 6.2%, 7.9% and 6.2% for the same analytes. The proposed approach has proved to be efficient, simple, easily transferable to industrial scale, cheap, fast and environmentally friendly.

Chlorine↗

Determination of the oxidative stability of olive oil, using focused-microwave energy to accelerate the oxidation process.

A new method is proposed for rapid monitoring of the oxidative stability of virgin olive oil. Samples were irradiated with microwave energy to accelerate the oxidation process and photometric monitoring was performed at 232 and 270 nm. Conditions such as microwave irradiation power, number of irradiation cycles, and irradiation time were optimized by means of multivariate screening that showed that irradiation power is the most significant condition in the oil oxidation process. Twelve samples of extra virgin olive oil of different oxidative stability--between 19 and 130 h calculated from the induction time of the Rancimat method, usual for analysis of olive oil in routine laboratories--were analyzed by the proposed microwave-assisted method, and a decrease in the induction time between 60 and 68% was obtained. The results obtained showed that correlation between the proposed method and the Rancimat method was excellent. The correlation coefficients were 0.9953 and 0.9963 for monitoring at 232 and 270 nm, respectively.

Microwaves↗

Ultrasound-assisted extraction of nitropolycyclic aromatic hydrocarbons from soil prior to gas chromatography-mass detection.

Continuous ultrasound-assisted extraction of nitropolycyclic aromatic hydrocarbons from soil prior to their individual separation and determination by gas chromatography (GC) with MS-MS detection is presented here. A multivariate optimisation of the variables affecting the continuous extraction step (namely, probe position, ultrasound radiation amplitude, percentage of duty cycle of ultrasonic exposure, sonication time, total extractant volume, extractant flow rate and temperature of the water-bath in which the extraction cell was placed) was performed. The method was compared with the reference EPA method 3540 using natural contaminated soils. Similar efficiencies were obtained but with a drastic reduction of both the extraction time (10 min versus 24 h) and the extractant volume (less than 10 ml versus 100 ml) by the proposed method. Detection limits of low picogram were obtained, with repeatability and reproducibility between 4.21-5.70 and 5.20-7.23%, respectively.

Gas Chromatography-Mass Spectrometry↗

Flow-injection spectrophotometric determination of cyanate in bioremediation processes by use of immobilised inducible cyanase.

A new flow injection (FI) method for photometric monitoring of cyanate in bioremediation processes using immobilised native cyanase is described. The method is based on the catalytic reaction between cyanate and bicarbonate to produce ammonia and carbon dioxide in the presence of an inducible native cyanase, immobilised in a reactor packed with glass beads. Two degrees of purification of the biocatalyst were used-heated cell-free extract and purified extract of cyanase from Pseudomonas pseudoalcaligenes CECT 5344. The ammonia produced by the enzymatic reaction is finally monitored photometrically at 700 nm using a modification of the conventional Berthelot method. The method furnishes different calibration curves depending on the degree of purification of the cyanase, with linear ranges between 1.23 and 616.50 micromol L(-1) ( r(2)=0.9979, n=7) and between 1.07 and 308.25 micro mol L(-1) ( r(2)= 0.9992, n=7) for the heated cell-free extract and the purified cyanase extract, respectively. No statistically significant differences between the samples were found in the precision study evaluated at two cyanate concentration levels using one-way analysis of variance. A sampling frequency of 15 h(-1) was achieved. The method was used to monitor cyanate consumption in a cyanate bioremediation tank inoculated with Pseudomonas pseudoalcaligenes CECT 5344 strain. The correlation between cyanate degradation and ammonia production was tested using a conventional method. Finally, the method was applied to different samples collected from the bioremediation tank using the standard addition method; recoveries between 85.9 and 97.4% were obtained.

Biodegradation, Environmental↗

Superheated liquids for extraction of solid residues from winemaking processes.

Solid residues from winemaking processes have been subjected to extraction with superheated water-ethanol mixtures. Identification and characterization of the extracted compounds were achieved by spectrophotometry, gas chromatography with either flame-ionization or mass detectors, and-high performance liquid chromatography with UV detection. Extraction was performed statically with single or repeated cycles. All variables affecting the extraction process have been studied and optimised. The extraction time and temperature were 65 min and 210 degrees C, respectively. Extracts comprised two phases--an aqueous phase, rich in phenolic compounds, and an oily phase, comprising mainly fatty acids. The method allows manipulation of extract composition by changing the applied pressure, temperature, water-to-ethanol ratio, and pH. The method is faster than traditional extraction procedures for obtaining valuable compounds from these residues.

Journal Article↗

Comparison of the static, dynamic and static-dynamic pressurised liquid extraction modes for the removal of nitrated polycyclic aromatic hydrocarbons from soil with on-line filtration-preconcentration.

The three operational modes of pressurised liquid extraction (PLE) (namely, static, dynamic and static-dynamic) have been applied for the extraction of nitrated polycyclic aromatic hydrocarbons from both spiked and natural contaminated soils. A comparison of the three modes in terms of experimental set-up used, extraction time needed for total removal of the analytes and precision has been carried out. The use of a flow-injection manifold as interface between every pressurised extractor and a filtration-preconcentration system has allowed the partial automation of the proposed approaches. Efficiencies close to 100% have been provided by the three operational modes. However, the static-dynamic mode has been proved as the most suitable alternative providing the shortest extraction time (25 min) versus the static (30 min) and the dynamic (50-70 min) modes. Gas chromatography with MS-MS ion preparation mode has been used providing both high selectivity (no interferences were observed) and sensitivity (detection limits of low pg). The comparison of the proposed approaches with the reference method 3540 of the US Environmental Protection Agency (EPA) has shown that both methods provide similar efficiencies with an important shortening in the extraction time (25-70 min by PLE versus 24 h by the EPA method). The use of water as leaching agent has avoided the use of organic solvents providing an environmentally friendly method.

Filtration↗

New methods for acceleration of meat sample preparation prior to determination of the metal content by atomic absorption spectrometry.

Focused microwave-assisted digestion and ultrasound leaching have been applied for the extraction of Pb, Cd, Cr, Cu, Fe, Zn, Ca, and Mg from raw meat. Semimembranous muscle (SM) of raw pig ham was used for optimizing both the digestion and extraction steps by multivariate approaches. The detection and quantification limits were 0.5 and 0.9 microg kg(-1) for Pb, 0.06 and 0.1 microg kg(-1) for Cd, 0.2 and 1.2 microg kg(-1) for Cr, 0.4 and 3 microg kg(-1) for Cu, 0.04 and 0.1 mg kg(-1) for Fe, 0.012 and 0.017 mg kg(-1) for Zn, 0.3 and 0.4 mg kg(-1) for Ca, and 0.01 and 0.03 mg kg(-1) for Mg. The precision, expressed as relative standard deviation (RSD), ranged between 2.5 and 9.6% for focused microwave-assisted digestion and between 3.5 and 10.6% for ultrasound leaching. The methods were then compared with a reference method and applied to a certified reference material (bovine muscle 184, from the BCR). The t-test, applied to the results obtained from focused microwave-assisted digestion, revealed that they are in agreement (p>0.01) with the certified and estimated values in the case of Pb, Cd, Cr, Cu, Fe, Ca, Mg, and Zn but not in that of Fe. In the case of ultrasound leaching, only the extraction of Pb, Cu, and Ca was quantitative. The method based on microwave digestion provides more accurate and precise results than ultrasound leaching. These new procedures have many advantages with regards to conventional methods, namely, reduction of the extraction time, simplification of the process, avoidance of chemical emissions to the atmosphere, and no losses of metals by volatilization.

Animals↗

Automated method for the determination of vitamin D3 hydroxymetabolites in serum.

An almost automated method for the determination of hydroxymetabolites of vitamin D(3) (cholecalciferol) in human serum is reported. The method consists of three steps: 1) a batch liquid-liquid extraction step with 2-propanol and hexane, and drying of the extract and reconstitution with phosphate buffer. 2) A cleanup and preconcentration step based on solid-phase extraction using Prospekt equipment, with CN group cartridges and elution with the chromatographic mobile phase. 3) A chromatographic step for individual separation of the target analytes starting with a 90:10 methanol-water mixture, then a linear gradient to obtain 100% methanol; followed by photometric detection. The method provides a linear range between 1.0 and 100 ng mL(-1) for 24,25-(OH)(2) vitamin D(3) and for 25-(OH)(2) vitamin D(3), and between 1.5 and 100 ng mL(-1) for 1,25-(OH) vitamin D(3), with correlation coefficients ranging between 0.993 and 0.987, repeatability between 1.9% and 4.8% and within-laboratory reproducibility between 2.8% and 8.8%.

Automation↗

Continuous ultrasound-assisted extraction coupled to on line filtration-solid-phase extraction-column liquid chromatography--post column derivatisation-fluorescence detection for the determination of N-methylcarbamates in soil and food.

A dynamic ultrasound-assisted method for the extraction of N-methylcarbamates (oxamyl, dioxacarb, metolcarb, carbofuran, carbaryl and isoprocarb) from soils and foods is proposed. The main factors affecting the extraction efficiency have been optimised by means of a central composite design. Pure water can be used as leaching agent. A flow injection manifold coupled to the extractor allows automation of the several steps involved in the analytical process. The method allows extraction of the carbamates from soil and food at 1 microg/g spiked level, with recoveries similar to those provided by the EPA 8318 method, without degradation of the target compounds during the extraction and with drastic shortening of the time required for this step (2 min vs. 4 h). Recoveries of the target analytes were 77-95% for spiked soil and 85-101% for spiked food. The detection and quantification limits were 12 and 40 ng/g, respectively, for all analytes, except carbaryl (detection and quantification limits 3 and 10 ng/ng, respectively). The relative standard deviations for repeatability and within-laboratory reproducibility were 3.1 and 7.5%, respectively.

Carbamates↗

Automated fast extraction of nitrated polycyclic aromatic hydrocarbons from soil by focused microwave-assisted Soxhlet extraction prior to gas chromatography--electron-capture detection.

An approach for the automated fast extraction of nitrated polycyclic aromatic hydrocarbons (nitroPAHs) from soil, using a focused microwave-assisted Soxhlet extractor, is proposed. The main factors affecting the extraction efficiency (namely: irradiation power, irradiation time, number of cycles and extractant volume) were optimised by using experimental design methodology. The reduction of the nitro-PAHs to amino-PAHs and the derivatisation of the reduced analytes with heptafluorobutyric anhydride was mandatory prior to the separation-determination step by gas chromatography--electron-capture detection. The proposed approach has allowed the extraction of these pollutants from spiked and "real" contaminated soils with extraction efficiencies similar to those provided by the US Environmental Protection Agency methods 3540-8091, but with a drastic reduction in both the extraction time and sample handling, and using less organic solvent, as 75-85% of it was recycled.

Chromatography, Gas↗

Rapid analytical method for the determination of pesticide residues in sunflower seeds based on focused microwave-assisted Soxhlet extraction prior to gas chromatography-tandem mass spectrometry.

A rapid analytical method for determination of organochlorine pesticide residues in sunflower seeds based on focused microwave-assisted Soxhlet extraction has been developed. The main factors affecting the extraction efficiency--namely microwave power, irradiation time, volume of extractant and number of cycles--were optimized by a two-level factorial fractional design. After extraction, a liquid-liquid extraction and a clean-up step including the use of Florisil macrocolumns were required prior to injection of the extracts into the chromatograph in order to isolate the pesticide residues from the lipid fraction of the original extract. The MS-MS ion preparation mode was selected due to the high sensitivity and selectivity it provides. Seed samples were used collected near a crop subjected to aerial pesticide application. Residues of hexachlorocyclohexane isomers and endosulfan were found in the seeds although they were not subjected to pesticide application, thus showing the spray-drift contamination. The validation of the proposed approach was carried out by comparison with the ISO 659-1988 reference extraction method obtaining similar, or even better efficiencies by the proposed approach.

Gas Chromatography-Mass Spectrometry↗

Superheated water extraction of cholesterol from solid food.

A method based on superheated water extraction has been developed for the removal of cholesterol from solid food thus providing a clean approach by avoiding the use of organic solvents. A preconcentration step was also studied with the aim of solving the problem derived from low-cholesterol-content samples and dilution effect. The research also involved the optimisation of the parameters affecting the extraction process by a central composite experimental design as well as a univariate study of the optimisation of the preconcentration step. The time required for total removal of the target compound was 60 min. The method was validated using a certified reference material (NIST-CRM 1845) and was used to analyse food samples within a wide range of cholesterol concentrations. The efficiency for the CRM was 105%. The precision of the method yielded values less than 6.5% (expressed as relative standard deviation) in all instances.

Chemical Fractionation↗

Enzyme inhibition-based biosensors and biosensing systems: questionable analytical devices.

An overview of methods based on enzyme inhibition is presented. Both biosensors and biosensing systems (implemented in continuous and discontinuous approaches) are considered. The aim of the overview is to alert users on the use of these devices which, despite they can involve selective biocatalysts, the inhibition effect is not selective. Tables and examples illustrate this assertion.

Biosensing Techniques↗

Static pressurised liquid extraction of nitrated polycyclic aromatic hydrocarbons from soils with on-line filtration-preconcentration prior to gas chromatography-mass spectrometry detection.

Nitrated polycyclic aromatic hydrocarbons have been extracted from both spiked and natural contaminated soils by using static pressurised liquid extraction. The variables governing the extraction (namely, pressure, extraction time, temperature and number of static extraction cycles) have been optimised using the experimental design methodology. A laboratory-made pressurised liquid extractor has been used allowing its coupling to a filtration-preconcentration system through a flow injection interface, thus providing a partially automated approach. Gas chromatography with MS-MS ion preparation mode has been used for individual separation-identification/quantification providing both high selectivity (no interferences were observed) and sensitivity (detection limits of low pg). The comparison of the proposed approach with the reference EPA Method 3540 has shown that both methods provide similar efficiencies but with dramatic saving of time by the former (30 min vs. 24 h). The use of water as leaching agent avoids the use of organic solvents, thus providing an environmentally friendly method.

Filtration↗