Elution Of Analytes Research Articles

Preparation of cyano‐functionalized multiwalled carbon nanotubes as solid‐phase extraction sorbent for preconcentration of phenolic compounds in environmental water

In the present work, we showed a novel method to synthesize cyano-functionalized multiwalled carbon nanotubes (MWCNTs-CN) and utilize it as a solid-phase extraction sorbent for preconcentration of phenolic compounds in environmental water samples. MWCNTs-CN was synthesized through surface functionalization of multiwalled carbon nanotubes (MWCNTs). The functional groups on the surface of modified MWCNTs were characterized by Fourier transform infrared spectroscopy, thermogravimetric analysis and scanning electron microscopy. The analytical procedure was based on a conventional solid-phase extraction step for which 100 mg of MWCNTs-CN were packed in a 3 mL polypropylene cartridge. Analytes were thus isolated and preconcentrated from the pretreated samples and subsequently detected on high-performance liquid chromatography-ultraviolet detection. The results showed the proposed method exhibited good sensitivity and precision for the extraction and elution of analytes. The limit of detections (S/N = 3) of the method were 0.45, 0.09, 0.08, and 3.00 ng mL(-1) for p-chlorophenol, 1-naphthol, 2-naphthol, and 2,4-dichlorophenol, respectively. The mean relative recoveries (n = 3) were between 80.28 and 103.13%, and the repeatability (RSD ≤ 5.10%) and reproducibility (RSD ≤ 7.68%) were accepted. This developed method was applied to determine phenolic compounds in environmental water samples. There is a positive result only for 2-naphthol with concentration of 0.38 ng mL(-1) in seawater sample.

Extraction and stability of pesticide multiresidues from natural water on a mixed-mode admicellar sorbent

The suitability of a mixed-mode sorbent made up of admicelles of sodium dodecyl sulphate (SDS) and tetrabutylammonium (TBA) to extract and preserve pesticides from river and underground water was assessed. Pesticides belonging to different structural groups (i.e. triazines, carbamates, phenylureas, anilides, chloroacetanilides, organophosphorus and phenoxyacids), most of them well known by their instability, were selected for this purpose. Extraction of pesticides with the admicellar sorbent was optimized. Percolation of 250mL of sample through the SPE cartridge, elution with 1mL of methanol (neutral and basic pesticides) or 2mL of 0.3M NaOH:methanol (90:10, v/v) (acidic pesticides) and direct analysis of the extract by liquid chromatography/UV detection, permitted to obtain method detection limits in the range 2–60ngL−1. Their degradation on the SPE cartridges after sample percolation was investigated under a variety of experimental conditions, namely darkness, natural illumination and different temperatures (room temperature, 4°C and −20°C) for 3 months. Under darkness, most of pesticides were stable for 1 month at room temperature and 3 months at 4°C. Only atriazine and clorfenvinfos did not follow this general behaviour. No influence of matrix components on the stabilization of pesticides in the admicellar sorbent was observed. The stabilizing capability of surfactant aggregates surpassed that of C18 and black carbon and was similar to that obtained in some polymeric materials, which have the disadvantage of requiring large volumes of solvents for analyte elution. The stabilization of pesticides on the admicellar sorbent was long enough to permit easy shipping and storage of the cartridges in monitorization campaigns.

Chromatographic models to predict the elution of ionizable analytes by organic modifier gradient in reversed phase liquid chromatography

The retention of an ionizable analyte under RP-HPLC organic modifier gradient elution is strongly affected by its ionization degree which, in turn, depends on its pKa and on the pH of the mobile phase. The values of both parameters change depending on the mobile phase composition and thus retention becomes a parameter quite difficult to predict, particularly when working in gradient mode. In this work, an equation describing the retention of ionizable analytes has been combined with three different models of different complexity, developed for gradient elution of neutral compounds (1, 2, or 3 fitting parameters), in order to predict retention of compounds with acid–base properties with particular buffers. All models have been tested under 16 different gradient patterns (4 linear gradients, 4 concave gradients, 4 convex gradients and 4 combinations between them) for the prediction of the retention time of 12 acid–base compounds (pKa values from 4 to 9) in 3 different buffered mobile phases (pH 5, pH 7 and pH 9) with acetonitrile as organic modifier. The agreement between the experimental and calculated retention times is good for all models. The best results are obtained through the model that depends on three parameters and the accuracy of the two-parameter model is slightly lower but very acceptable too. On the other hand, the predictions performed with the one-parameter model are the less accurate, but good enough to become a valid model taking into account that it requires very little experimental work.

Cyclodextrins as a chiral mobile phase additive in nano-liquid chromatography: comparison of reversed-phase silica monolithic and particulate capillary columns

Enantioseparations of racemic nonsteroidal anti-inflammatory drugs (naproxen, ibuprofen, ketoprofen, flurbiprofen, suprofen, indoprofen, cicloprofen, and carprofen) were performed by nano-liquid chromatography, employing achiral capillary columns and heptakis(2,3,6-tri-O-methyl)-β-cyclodextrin (TM-β-CD) or hydroxylpropyl-β-cyclodextrin (HP-β-CD) as a chiral mobile phase additive (CMPA). Working under the same experimental conditions (in terms of mobile phase and linear velocity), the performance of a RP-C18 monolithic column was compared with that of a RP-C18 packed column of the same dimensions (100 μm i.d. × 10 cm). Utilizing a mobile phase composed of 30% ACN (v/v) buffered with 50 mM sodium acetate at pH 3, and containing 30 mM TM-β-CD, the monolithic column provided faster analysis but lower resolution than the packed column. This behavior was ascribed to the high permeability of the monolithic column, as well as to its minor selectivity. HP-β-CD was chosen as an alternative to TM-β-CD. Employing the monolithic column, the effects of different parameters such as HP-β-CD concentration, mobile phase composition, and pH on the retention factor and the chiral resolution of the analytes were studied. For the most of the analytes, enantioresolution (which ranged from R(s) = 1.80 for naproxen to R(s) = 0.86 for flurbiprofen) was obtained with a mobile phase consisting of sodium acetate buffer (25 mM, pH 3), 10% MeOH, and 15 mM HP-β-CD. When the same experimental conditions were used with the packed column, no compound eluted within 1 h. Upon increasing the percentage of organic modifier to favor analyte elution, only suprofen eluted within 30 min, with an R(s) value of 1.14 (20% MeOH). Replacing MeOH with ACN resulted in a loss of enantioresolution, except for naproxen (R(s) = 0.89).

A diffusive sampling device for measurement of ammonia in air

Abstract A diffusive sampling device, the DSD-NH3, has been developed for measurement of ammonia in air. The DSD-NH3 comprises silica gel impregnated with phosphoric acid as the absorbent, a porous sintered polyethylene tube that acts as a diffusive membrane, and a small polypropylene syringe that is used for the elution of analytes from the absorbent. Silica gel impregnated with phosphoric acid is used as absorbent for the DSD-NH3; basic gases in ambient air, including ammonia, are trapped in the DSD-NH3 device by their reaction with phosphoric acid in the sampler to form their corresponding phosphoric acid salts. After collection, the DSD-NH3 samplers are eluted by water. Cations in the eluate, including ammonium ions, are analyzed by ion chromatography. A side-by-side comparison was made with active samplers, demonstrating good correlation (r2 = 0.996). The sampling rate (94.5 ml min−1) was determined from comparison with an active sampling method and sampling rates. The sampling rate is also calculated from the respective molecular weights according to a rule based on Graham’s law. The theoretical sampling rate with the DSD-NH3 is 95.4 ml min−1 and agrees with the experimental value (94.5 ml min−1). Little influence of wind velocity on the sampler was observed. The relative standard deviations for ammonia concentrations were 4.7% with face velocity ranging 0–5.0 m/s.

Microfluidic droplet-array liquid–liquid chromatography based on droplet trapping technique

We describe the first realization of liquid chromatographic separation in a droplet-based microfluidic system and develop a novel mode for microchip-based chromatography named as droplet-array liquid-liquid chromatography. In this system, two arrays of picoliter-scale droplets immobilized on both sidewalls of a microchannel with droplet trapping technique served as the stationary phase in chromatographic separation, while the other immiscible phase flowing in the microchannel served as the mobile phase. The chromatographic separation was achieved on the basis of multiple extraction and elution of analytes between the droplet array stationary phase and the mobile phase. The proof-of-concept study of the droplet-array LC system was performed in the separation of fluoranthene and benzo[b]fluoranthene. Under the optimum conditions, the two analytes were separated within 26 min with separation efficiencies of 112 μm and 119 μm plate height, respectively. The advantages of the present system include simple structure, low driving pressure, and relatively high sample capacity. It can also provide a useful platform for LC theory study and educational purposes by allowing the researchers and students to directly "see" the continuous extraction and elution process of a chromatographic separation.

Characterization of hexacationic imidazolium ionic liquids as effective and highly stable gas chromatography stationary phases

Polycationic ionic liquids (ILs) are an attractive class of ILs with great potential applicability as gas chromatography stationary phases. A family of hexacationic imidazolium ILs derived from the cycloalkanol family was chemically first prepared in a straightforward manner and then applied for analytical separation purposes. Four tuneable engineering vectors, namely cation ring size structure, anion nature, spatial disposition of cycloalkanol substituents and O-substitution, were considered as experimental parameters for the design of the desired ionic liquids. A total number of five new phases based on a common benzene core respectively exhibited column efficiencies around to 2500 plates/m, broad operating temperature ranges and also, even more importantly, good thermal stabilities (bleeding temperature between 260 and 365°C), finding variations in the selectivity and analytes elution orders depending on the IL structures. Their solvation characteristics were evaluated using the Abraham solvation parameter model, establishing clear correlations between their cation structure and retention capability with respect to certain analytes. The study of relationships between the ILs structure and solvation parameters gives us an idea of the IL stationary phase to be used for specific separations.

Single-Electrolyte Isotachophoresis Using a Nanochannel-Induced Depletion Zone

Isotachophoretic separations are triggered at the border of a nanochannel-induced ion-depleted zone. This depletion zone acts as a terminating electrolyte and is created by concentration polarization over the nanochannel. We show both continuous and discrete sample injections as well as separation of up to four analytes. Continuous injection of a spacer compound was used for selective analyte elution. Zones were kept focused for over one hour, while shifting less than 700 μm. Moreover, zones could be deliberately positioned in the separation channel and focusing strength could be precisely tuned employing a three-point voltage actuation scheme. This makes depletion zone isotachophoresis (dzITP) a fully controllable single-electrolyte focusing and separation technique. For on-chip electrokinetic methods, dzITP sets a new standard in terms of versatility and operational simplicity.

Evaluating the solvation properties of functionalized ionic liquids with varied cation/anion composition using the solvation parameter model

Ionic liquids (ILs) are promising gas chromatography (GC) stationary phases due to their high thermal stability, negligible vapor pressure, and ability to solvate a broad range of analytes. The tunability of ILs allows for structure modification in pursuit of enhanced separation selectivity and control of analyte elution order. In this study, the solvation parameter model is used to characterize the solvation interactions of fifteen ILs containing various cationic functional groups (i.e., dimethylamino, hydroxyl, and ether) and cation types paired with various counter anions, namely, tris(pentafluoroethyl)trifluorophosphate (FAP −), bis[(trifluoromethyl)sulfonyl]imide (NTf 2 −), thiocyanate (SCN −), tricyanomethide (C(CN) 3 −), tetracyanoborate (B(CN) 4 −), and bis[oxalate(2-)]borate (BOB −). The presence of functional groups affected the hydrogen bond basicity, hydrogen bond acidity, as well as dispersion interactions of the resulting ILs, while the change of cation type yielded modest influence on the dipolarity. The switch of counter anions in unfunctionalized ILs produced compounds with higher dipolarity and hydrogen bond basicity. The dipolarity and hydrogen bond basicity of ILs possessing cyano-containing anions appeared to be inversely proportional to the cyano content of the anion. The modification of IL structure resulted in a significant effect on the retention behavior as well as separation selectivity for many solutes, including reversed elution orders of some analytes. This study provides one of the most comprehensive examinations up-to-date on the relation between IL structure and the resulting solvation characteristics and gives tremendous insight into choosing suitable ILs as GC stationary phases for solute specific separations.

Studies of ion-imprinted polymers for solid-phase extraction of ruthenium from environmental samples before its determination by electrothermal atomic absorption spectrometry

The examination of the effect of interfering ions on the analytical signal of ruthenium measured by electrothermal atomic absorption spectrometry was initially performed in this work. The complexes of ruthenium(III) with thiosemicarbazide (TSd) and acetaldehyde thiosemicarbazone (AcTSn) were prepared and imprinted in polymeric network. The ion-imprinted polymers were synthesized by copolymerization of methacrylic acid, as functional monomer and ethylene glycol dimethacrylate, as crosslinking agent in the presence of 2,2-azobisisobutyronitrile as initiator. The effects of sample volume, pH, and flow rate on the extraction of analyte were studied in dynamic mode. The optimum pH for quantitative retention of ruthenium on each of the studied sorbents was 7.5 ± 0.5. The elution of analyte was completed with 0.2 mol L −1 thiourea in 0.2 mol L −1 HCl. The effect of matrix ions on ruthenium(III) separation process was studied. The analytical performance of the Ru–TSd polymer in the presence of competing ions was better than Ru–AcTSn polymer, considering recovery of analyte, reproducibility of results, selectivity coefficients, and sorbent capacity. The detection limit of the proposed method (0.16 ng mL −1 on Ru–TSd and 0.25 ng mL −1 on Ru–AcTSn) is lower in comparison with the previously published methods. The developed separation method was successfully applied to the determination of trace amounts of ruthenium in spiked water samples, sludge, grass, and human hair.

Tandem use of solid‐phase extraction and dispersive liquid–liquid microextraction for the determination of mononitrotoluenes in aquatic environment

Solid-phase extraction (SPE) in tandem with dispersive liquid-liquid microextraction (DLLME) has been developed for the determination of mononitrotoluenes (MNTs) in several aquatic samples using gas chromatography-flame ionization (GC-FID) detection system. In the hyphenated SPE-DLLME, initially MNTs were extracted from a large volume of aqueous samples (100 mL) into a 500-mg octadecyl silane (C(18)) sorbent. After the elution of analytes from the sorbent with acetonitrile, the obtained solution was put under the DLLME procedure, so that the extra preconcentration factors could be achieved. The parameters influencing the extraction efficiency such as breakthrough volume, type and volume of the elution solvent (disperser solvent) and extracting solvent, as well as the salt addition, were studied and optimized. The calibration curves were linear in the range of 0.5-500 μg/L and the limit of detection for all analytes was found to be 0.2 μg/L. The relative standard deviations (for 0.75 μg/L of MNTs) without internal standard varied from 2.0 to 6.4% (n=5). The relative recoveries of the well, river and sea water samples, spiked at the concentration level of 0.75 μg/L of the analytes, were in the range of 85-118%.

Development of immunoaffinity columns for pyraclostrobin extraction from fruit juices and analysis by liquid chromatography with UV detection

Pyraclostrobin belongs to a new generation of fungicides widely used to preserve high valuable crops. In the present study, three monoclonal antibodies with different affinities to this modern strobilurin have been evaluated for their usefulness in the production of immunoaffinity columns suitable for the solid-phase extraction, concentration, and clean-up of residues from food commodities. Different immunosorbents were produced and characterized in terms of antibody immobilization efficiency, immunosorbent binding capacity, optimum elution conditions, and reusability. Covalent coupling of the antibodies to Sepharose–CNBr gel took place with high yield (over 90%), whereas the immunosorbent efficacy to retain the analyte (from 28 to 68%) was shown to depend on the amount and type of antibody immobilized on the support. As a matter of fact, columns prepared with the monoclonal antibody PYs5#14 were able to selectively bound up to 53 μg of pyraclostrobin per gram of beads. Acetonitrile solutions were preferred over methanolic ones for analyte elution, and some immunosorbents could be reused at least 4–6 times provided that the amount of pyraclostrobin and the volume of sample did not overload the column. Effectiveness of the selected immunoaffinity column was evidenced by the development of an extraction procedure for pyraclostrobin residues from fruit juices and further determination by high-performance liquid chromatography with UV detection. A concentration factor of 50 times was achieved with the developed immunoaffinity column, which eventually resulted in a limit of quantification of 0.01 mg L −1. Finally, quantitative recoveries were obtained on apple juice and red grape must samples spiked with pyraclostrobin from 0.01 to 1 mg L −1.

Development and validation of a highly sensitive LC-MS/MS method for simultaneous quantitation of acetyl-CoA and malonyl-CoA in animal tissues

A highly sensitive and specific LC-MS/MS method was developed for simultaneous estimation of acetyl co-enzyme A (ACoA) and malonyl co-enzyme A (MCoA) in surrogate matrix using n-propionyl co-enzyme A as an internal standard (IS). LC-MS/MS was operated under the multiple reaction-monitoring mode using the electrospray ionization technique. Simple acidification followed by dilution using an assay buffer process was used to extract ACoA, MCoA and IS from surrogate matrix and tissue samples. The total run time was 3 min and the elution of both analytes (ACoA, MCoA) and IS occurred at 1.28 min; this was achieved with a mobile phase consisting of 5 mM ammonium formate (pH 7.5)-acetonitrile (30:70, v/v) delivered at a flow rate of 1 mL/min on a monolithic RP-18e column. A linear response function was established for the range of concentrations 1.09-2187 and 1.09-2193 ng/mL for ACoA and MCoA, respectively. The intra- and inter-day precision values for ACoA and MCoA met the acceptance as per FDA guidelines. ACoA and MCoA were stable in a battery of stability studies viz. bench-top, auto-sampler and long-term. The developed assay was used to quantitate ACoA and MCoA levels in various tissues of rat.

Toward the use of surface modified activated carbon in speciation: Selective preconcentration of selenite and selenate in environmental waters

This paper describes a novel application of tetrabutylammonium hydroxide-modified activated carbon (AC-TBAH) to the speciation of ultra-trace Se(IV) and Se(VI) using LC-ICP-DRC-MS. The anion exchange functionality was immobilized onto the AC surface enables selective preconcentration of inorganic Se anions in a wide range of working pHs. Simultaneous retention and elution of both analytes, followed by subsequent analysis with LC-ICP-DRC-MS, allows to accomplish speciation analysis in natural samples without complicated redox pre-treatment. The laboratory-made column of immobilized AC (0.4 g of sorbent packed in a 6 mL syringe barrel) has achieved analyte enrichment factors of 76 and 93, respectively, for Se(IV) and Se(VI), thus proving its superior preconcentration efficiency and selectivity over common AC. The considerable enhancement in sensitivity achieved by using the preconcentration column has improved the method's detection limits to 1.9–2.2 ng L −1, which is a 100-fold improvement compared with direct injection. The analyte recoveries from heavily polluted river matrix were between 95.3 and 107.7% with less than 5.0% RSD. The robustness of the preconcentration and speciation method was validated by analysis of natural waters collected from rivers and reservoirs in Hong Kong. The modified AC material is hence presented as a low-cost yet robust substitute for conventional anion exchange resins for routine applications.

Microextração em sorvente empacotado (MEPS) para a determinação de fármacos em fluidos biológicos

Microextraction by packed sorbent (MEPS) is a new sample preparation technique developed in 2004. Actually, MEPS is a miniaturization of conventional solid phase extraction (SPE) packed bed devices from mililitre bed volumes to microlitre volumes. The key factor in MEPS is that the volume of solvent used to elute the analytes from the extraction process is of a suitable order of magnitude to be injected directly on-line into Liquid Chromatography, Gas Chromatography or Capillary Electrophoresis systems, without any modification of the instrument. This review focuses on MEPS theory and applications. MEPS procedure with different steps (sample preparation, analytes pre-concentration, sorbent washing, and analytes elution) is discussed for biological samples. A short overview of recent and successful applications of MEPS for determination of drugs and their metabolites in different biological matrices (plasma, urine, blood, hair) for therapeutic drug monitoring, doping inspection, biomarkers analysis, and forensic toxicology are also summarized.

Synthesis, Spectral Assignment and Application of a New Synthesized Schiff Base Ionophore to Determination of Ultra Trace Copper by Solid Phase Extraction Flame Atomic Absorption Spectrometry

The present report deals with a very selective, facile and precise method for rapid separation of the trace copper in aqueous samples using octadecyl silica bonded phase membrane disks modified by N,N’-diethylethylendiaminyl,2,6-pyridinedicarboxylate (DDPD) combined with flame atomic absorption spectrometric determination. Besides, synthesis and spectral characterization of DDPD have been described in detail. All the affecting experimental variables such as pH, amount of modifier, eluent type, sample and eluent flow-rate, interfering ions and disk capacity were also investigated. The target analyte (trace copper) was quantitatively retained at pH = 4, and eluted with 6 ml of HNO3 0.5 M applying flow rates of 45 ml min−1 and 12 ml min−1 for analyte passage and elution steps, respectively. The proposed method also permits an enrichment factor of about 500 and a detection limit of 0.005 ng ml−1. The method has been successfully applied for isolation and determination of copper in different aqueous, real samples, peppers and standard alloys.

LC-MS/MS method for the determination of carbamathione in human plasma

Liquid chromatography-tandem mass spectrometry methodology is described for the determination of S-(N,N-diethylcarbamoyl)glutathione (carbamathione) in human plasma samples. Sample preparation consisted of a straightforward perchloric acid medicated protein precipitation, with the resulting supernatant containing the carbamathione (recovery ∼98%). For optimized chromatography/mass spec detection a carbamathione analog, S-(N,N-di-i-propylcarbamoyl)glutathione, was synthesized and used as the internal standard. Carbamathione was found to be stable over the pH 1–8 region over the timeframe necessary for the various operations of the analytical method. Separation was accomplished via reversed-phase gradient elution chromatography with analyte elution and re-equilibration accomplished within 8min. Calibration was established and validated over the concentration range of 0.5–50nM, which is adequate to support clinical investigations. Intra- and inter-day accuracy and precision determined and found to be <4% and <10%, respectively. The methodology was utilized to demonstrate the carbamathione plasma-time profile of a human volunteer dosed with disulfiram (250mg/d). Interestingly, an unknown but apparently related metabolite was observed with each human plasma sample analyzed.

A validated HPLC method for simultaneous determination of four active components in counter cough formulation

A new quantitative reversed-phase HPLC method has been developed for the simultaneous determination of phenylephrine hydrochloride (PEH), guaiphenesin (GP), ambroxol hydrochloride (AMBH) and levocetirizine dihydrochloride (LCZH) in counter cough formulation. Mobile phase consists of phosphate buffer of pH 3.0 and solvent mixture (methanol: acetonitrile in the ratio of 1: 2) using gradient system. Column reversed phase Luna C 8 (5 μm, 250 x 4.6 mm i.d.) phenomenex is used as stationary phase. The elution of analytes is achieved in less than 30 min with a flow rate of 1.0 ml min -1 . Detection is carried out using a variable wavelength UV-Vis detector at 273 nm and 230 nm. Linearity and percent recoveries for PEH, GP, AMBH and LCZH where R 2 > 0.999 and 100.18, 99.92, 100.26 and 100.22% respectively. Therefore, the reported method is selective, specific, precise and accurate for the analyses of different constituents in drug substance and pharmaceutical dosage form.

Quantification of theobromine and caffeine in saliva, plasma and urine via liquid chromatography–tandem mass spectrometry: A single analytical protocol applicable to cocoa intervention studies

Targeted analyses of clinically relevant metabolites in human biofluids often require extensive sample preparation (e.g., desalting, protein removal and/or preconcentration) prior to quantitation. In this report, a single ultra-centrifugation based sample pretreatment combined with a designed liquid chromatography–tandem mass spectrometry (LC–MS/MS) protocol provides selective quantification of 3,7-dimethylxanthine (theobromine) and 1,3,7-trimethylxanthine (caffeine) in human saliva, plasma and urine samples. The optimized chromatography permitted elution of both analytes within 1.3 min of the applied gradient. Positive-mode electrospray ionization and a triple quadruple MS/MS instrument operated in multiple reaction mode were used for detection. 13C 3 isotopically labeled caffeine was included as an internal standard to improve accuracy and precision. Implementing a 20-fold dilution of the isolated low MW biofluid fraction prior to injection effectively minimized the deleterious contributions of all three matrices to quantitation. The assay was linear over a 160-fold concentration range from 2.5 to 400 μmol L −1 for both theobromine (average R 2 0.9968) and caffeine (average R 2 0.9997) respectively. Analyte peak area variations for 2.5 μmol L −1 caffeine and theobromine in saliva, plasma and urine ranged from 5 and 10% (intra-day, N = 10) to 9 and 13% (inter-day, N = 25) respectively. The intra- and inter-day precision of theobromine and caffeine elution times were 3 and <1% for all biofluids and concentrations tested. Recoveries for caffeine and theobromine ranged from 114 to 118% and 99 to 105% at concentration levels of 10 and 300 μmol L −1. This validated protocol also permitted the relative saliva, plasma and urine distribution of both theobromine and caffeine to be quantified following a cocoa intervention.

An Aptameric Microfluidic System for Specific Purification, Enrichment, and Mass Spectrometric Detection of Biomolecules

We present an innovative microfluidic system that accomplishes specific capture, enrichment, and isocratic elution of biomolecular analytes with coupling to label-free mass spectrometric detection. Analytes in a liquid phase are specifically captured and enriched via their affinity binding to aptamers, which are immobilized on microbeads packed inside a microchamber. Exploiting thermally induced reversible disruption of aptamer-analyte binding via on-chip temperature control with an integrated heater and temperature sensor, the captured analytes are released into the liquid phase and then isocratically eluted and transferred via a microfluidic flow gate for detection by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The utility of the device is demonstrated using adenosine monophosphate (AMP) as a model analyte. Experimental results indicate that the device is capable of purifying and enriching the analyte from a sample mixed with nonspecific analytes and contaminated with salts. In addition, thermally induced analyte release is performed at modest temperatures (45degC), and mass spectra obtained from MALDI-MS demonstrate successful detection of AMP at concentrations as low as 10 nM following enrichment by consecutive infusion of a diluted sample.