Precise evaluation of batch adsorption kinetics of plant total polyphenols based on a flow-injection online spectrophotometric method.

Efficient evaluation of adsorption kinetics of plant total polyphenols is essential for the design of adsorption separation of bioactive compounds. The conventional method uses manual sampling with poor reproducibility. Here, we developed a new method for on-line determination of total polyphenol content (TPC) in plant extracts by applying the Folin-Ciocalteu method in flow-injection analysis (FIA). The FIA parameters were optimized and a standard curve with excellent linearity was established. Precise determination of TPC with a satisfactory sample throughput of 20 h-1 was achieved for the adsorption kinetic study. The pseudo-second-order kinetic model was found to better describe the kinetic parameters of the batch adsorption/desorption process. The developed method proved to be accurate compared with the conventional method. The FIA method holds significant promise for studying and monitoring adsorption processes, due to its automatic on-line nature, low consumption of reagents and samples, and the ability to generate large quantities of highly accurate adsorption data.

Determination of Selenium in Selenium-Enriched Products by Specific Ratiometric Fluorescence.

Selenium (Se), as one of the essential and nutrient components of living organisms and plants, plays an important role in life activities, while excessive selenium is hazardous to human health. So, the establishment of an effective method for simple, rapid, and highly sensitive determination of selenium content is crucial in the field of food composition analysis and other areas. In this paper, a novel and simple ratiometric fluorescence method for the determination of Se has been developed using 9-anthracenemethanol (AM) as the ratiometric fluorescence reagent on the basis of the conventional fluorometric assay which utilized 2,3-diaminonapthalene (DAN) as fluorescent ligand. The ratiometric method was compared with the conventional method with respect to precision and accuracy. The inter-day and intra-day precisions (RSDs) of the ratiometric fluorescence method ranged from 2.08 to 2.78% and 1.28 to 1.84%, with mean recoveries of 93.2~98.0% and limit of detection (LOD) and limit of quantification (LOQ) of 0.0016 and 0.0049 µg/mL, respectively. This method was successfully applied to the determination of total selenium in selenium-enriched milk and selenium-supplemented shampoo, with the results in agreement with those obtained by inductively coupled plasma mass spectrometer (ICP-MS). The results demonstrated that the precision and accuracy of the ratiometric fluorescence method were superior to those of the conventional fluorescence method, and the interferences of various environmental factors were effectively eliminated. The precision and accuracy of the conventional method can be significantly improved by simply adding an elaborately selected ratiometric fluorescence reagent, and the new method will have broader practical applications.

A membrane-protected micro-solid-phase extraction method based on molecular imprinting and its application to the determination of local anesthetics in cosmetics.

As local anesthetics that are illegally added to cosmetics are harmful to consumer health, it is necessary to establish an efficient method for detecting these substances. Herein, a molecularly imprinted polymer (bupivacaine) was prepared by bulk polymerization and packed into a hollow fiber for use as an extraction phase to fabricate a membrane-protected micro-solid-phase extraction device. The optimal values of the influencing parameters for the microextraction process were as follows: a sample solution pH of 9.0, a loading and washing time of 2 h, and an elution time of 32 min. A gas chromatography-mass spectrometry method was established for the determination of local anesthetics and coupled with the microextraction method to successfully detect local anesthetics in cosmetic samples. The calibration curve for the proposed method was linear in the range of 0.4-50 mg/L and showed a good correlation coefficient (r2 ). The limits of detection for local anesthetics were in the range of 0.01-0.71 mg/L. The molecularly imprinted polymer exhibited good imprinting and selectivity, the micro-solid-phase extraction device was simple and inexpensive and fabrication was reproducible. The combination of molecular imprinting technology, membrane separation, and micro-solid-phase extraction methods used in this study can potentially be applied to pretreat local anesthetics in cosmetic samples.

Novel carrier-mediated membrane-assisted three-phase liquid-liquid extraction coupled with liquid chromatography-mass spectrometry for the determination of eight biogenic amines in foods.

Analysis of biogenic amines (BAs) is of great importance due to their toxicity and usage as indicators of food freshness. In this study, membrane-assisted three-phase liquid-liquid extraction (MA-3pLLE) was proposed to integrate extraction and back-extraction into a single step using a specially designed U-shaped device. Parameters affecting the performance of the extraction method were optimized. Coupled with liquid chromatography-mass spectrometry, the method was successfully applied for the simultaneous determination of eight BAs without derivatization in apple juice, orange juice, red wine, soy sauce and milk granules, with satisfactory recoveries and RSDs. The calibration curve of the method was linear in the range of 1.00~100.00 ng/mL, with a correlation coefficient (r2) ≥ 0.9908. The limits of detection were in the range of 0.03~1.00 ng/mL. MA-3pLLE is efficient, reproducible, and green and has great potential for application in the one-step extraction of analytes from complex matrices.

A molecularly imprinted polymer based monolith pipette tip for solid-phase extraction of 2,4-dichlorophenoxyacetic acid in an aqueous sample.

This paper presents a simple approach for fabrication of a pipette tip solid-phase extraction (PT-SPE) device, which possesses a monolith structure with low back pressure and has high selectivity to 2,4-dichlorophenoxyacetic acid (2,4-D). Pipette tips were packed with molecularly imprinted polymers (MIPs) as a selective adsorbent and high-density polyethylene (HDPE) as a co-sintering agent, and then heated to form a monolith extraction device. The key factors including the particle size and amount of packing material, and the type and volume of elution solvent, which influence PT-SPE device performance were optimized. A packing material of 40 mg/0.20 mL in a ratio of 4/6 (MIPs/HDPE) and treatment temperature of 150 °C was selected. By the determination with high-performance liquid chromatography (HPLC-SPD), the extraction device was found to have a good extraction recovery for a 2,4-D lake water sample at a low concentration (0.006 mg L-1) with an enrichment factor about 50. The proposed method provided a simple approach for the fabrication of a PT-SPE monolith device with reduced back pressure and wall effect, which are very important for improving the extraction efficiency. And the device will have promising application in the extraction of a variety of analytes in complex samples.

A simple on-line detection system based on fiber-optic sensing for the realtime monitoring of fixed bed adsorption processes of molecularly imprinted polymers.

Fixed bed adsorption is widely used for separations and purifications of active components in medicine, and for wastewater treatment. At present, fixed bed adsorption breakthrough curve is generally obtained by manual sampling and off-line detection. In this study, we proposed a method for on-line monitoring of fixed bed adsorption process using a self-assembled fiber-optic sensing (FOS) system. The adsorption of 2,4-dichlorophenoxyacetic acid (2,4-D) on the fixed bed packed with molecularly imprinted polymers (MIPs) and non-imprinted polymers (NIPs) were studied. The reproducibility and precision of the system was investigated. The relative standard deviation (RSD) of the system was less than 1.54%, which indicates that the system has a good reproducibility. The effects of initial concentration, flow rate, adsorbent mass and particle size on the breakthrough curves were investigated. Through screening, it was found that adsorption kinetics of the polymer materials fit to Thomas and Yoon-Nelson models. The MIPs showed high binding capacity, good selectivity, fast adsorption rate, indicating a great potential for the treatment of 2,4-D contaminated water. Moreover, this study has identified that the detection method has the advantages of being on-line, realtime, simple, and accurate. The on-line method can facilitate the study of fixed bed adsorption processes and accelerate the understanding of adsorption kinetics.

In-situ preparation of porous monolithic polymer inside hollow fiber as a micro-solid phase extraction device for glucocorticoids in cosmetics.

Glucocorticoids have a certain whitening effect on the skin. However, frequent and long-term use of cosmetics including glucocorticoids is harmful to health. Herein, we proposed a novel micro-solid phase extraction method for the detection of prednisolone acetate, prednisone, and prednisolone in cosmetics coupled with high-performance liquid chromatography. In this method, porous monolithic polymer micro-extraction bars were prepared by "one-step, one-pot" in situ photopolymerization combined with sacrificial support in hollow fiber under water atmosphere. The crucial factors such as pH of sample solution, extraction, and elution times that influence micro-extraction were optimized and found to be 9.0, 2 h, and 32 min, respectively. Under the optimum experimental conditions, the linear range of the calibration curves were from 5.0 to 2000 µg/L with correlation coefficients (R2 ) between 0.9922 and 0.9996. The limit of detection and limit of quantification were 1.5 µg/L and 5.0 µg/L, respectively, and the recoveries were found to be in range of 69.0-113.3%. The analysis of precision for intraday and interday were less than 10.40 and 10.59%. The device has been successfully achieved photopolymerization under water atmosphere. The results indicated that this method is simple, accurate, and satisfactory for the pretreatment and determination of glucocorticoids in complex cosmetics samples.

Membrane-Protected Molecularly Imprinted Polymer for the Microextraction of Indole-3-butyric Acid in Mung Bean Sprouts.

Based on the hollow fiber protected molecularly imprinted polymer, a micro-solid-phase extraction (µ-SPE) method was developed and applied for the analysis of indole-3-butyric acid in mung bean sprouts by high-performance liquid chromatography. The extraction conditions of the µ-SPE method were optimized using L9(34) orthogonal, and optimum conditions were found as follows: pH of sample solution was 2.0, chloroform was the organic solvent for embedding the µ-SPE bars, and acetonitrile was the desorption solvent. In addition, the extraction time was 80 min, desorption time was 5 min, stirring speed was 800 rpm, and concentration of NaCl was 10%. Under the optimum conditions, a standard curve was established for IBA, with a correlation coefficient of 0.9999. After extraction with phosphate buffer solution (pH = 9.0), successful pretreatment of mung bean sprouts was achieved by the µ-SPE method. The limit of detection was 0.075 mg/kg, and the recoveries were found to be in the range of 88.9-106.4%. This method is simple, environmentally friendly, and can be used for the determination of indole auxin contents in green bean sprouts quickly and accurately.

Preparation of a stoichiometric molecularly imprinted polymer for auramine O and application in solid-phase extraction.

We describe a stoichiometric approach to the synthesis of molecularly imprinted polymers specific for auramine O. Using the stoichiometric interaction in molecular imprinting, no excess of binding sites is necessary and binding sites are only located inside the imprinted cavities. The free base of the template was obtained to facilitate the interaction with the monomers. Itaconic acid was selected as the functional monomer, and stoichiometric ratio of the interaction with the free base was investigated. The molecularly imprinted polymer preparation conditions such as cross-linker, molar ratio, porogen were optimized as divinylbenzene, 1:2:20 and chloroform/N,N-dimethylformamide, respectively. Under the optimum conditions, a good imprinting effect and very high selectivity were achieved. A solid-phase extraction method was developed using the molecularly imprinted polymers as a sorbent and extraction procedure was optimized. The solid-phase extraction method showed a high extraction recovery for auramine O in its hydrochloride form and free form compared to its analogues. The results strongly indicated that stoichiometric imprinting is an efficient method for development of high selectivity molecularly imprinted polymers for auramine O.

Synthesis of fluorescent drug molecules for competitive binding assay based on molecularly imprinted polymers.

Fluorescent immunosorbent assay (FIA) is very promising for sensitive and selective analysis in bio-medical applications. Here, we proposed an assay, using fluorescent engineering of analytes and the corresponding molecularly imprinted polymers (MIPs) as a plastic antibody. Three drug molecules (metronidazole, zidovudine and lamivudine) were condensed with 9-aminoacridine, using succinic anhydride as a spacer. The target products were characterized with 1H-NMR, IR and mass spectrometry. UV-vis absorption and fluorescent properties of the fluorophore-labeled drug molecules were investigated. Feasibility of the fluorescent biomimetic immunosorbent assay based on MIPs was demonstrated in the solution. This work will provide sound foundation for the future application in real sample.

Preparation of stoichiometric molecularly imprinted polymer coatings on magnetic particles for the selective extraction of auramine O from water.

A novel magnetic molecularly imprinted polymer for the selective recognition of auramine O was rationally designed via screening from a library of nonimprinted polymers. A stoichiometric ratio of functional monomer (itaconic acid) and template molecule (auramine O) was found to be 1.5. Meanwhile, the synthesized SiO2 @Fe3 O4 was modified by 0.5 mol/L hydrochloric acid to facilitate the preparation of magnetic molecularly imprinted polymer particles. Adsorption experiments showed that the magnetic polymer particles exhibited good selectivity, recoveries, and enrichment performance. The stoichiometric imprinted polymers have been employed for the selective preconcentration of auramine O from lake water sample. The high specificity of the stoichiometric imprinted polymers was proven in the extraction of mixture solution of auramine O, auramine O hydrochloride, and chrysoidine, and the recoveries ranged between 99.66 and 108.75% (RSD 2.6-3.7%, n = 3) for lake water. These results suggest that this method is effective and can be successfully applied to the analysis of auramine O in environmental water samples.

In Situ Liquid-Phase-Adsorption Measurement System Based on Fiber-Optic Sensing with the Aid of Membranes.

At present, liquid phase adsorption (LPA) is still being quantitatively characterized in the way of manual sampling and off-line determination because of the complexity of the system comparing to gas adsorption. This paper describes a novel method for in situ, real-time measurement of LPA in general based on fiber-optic sensing (FOS) with the aid of membranes for the first time. A self-made measurement vessel was assembled from an adsorption bag, thermostatic devices with a stirrer, and a fiber-optic dipping probe. Also, macroporous adsorption resins (MARs) and rutin were chosen as model adsorbent and adsorbate to establish the FOS system. Here, in situ light absorption measurement was achieved by eliminating interference of adsorbent particles via encapsulating them with a membrane into the adsorption bag. In situ LPA measurement of rutin solution on MARs was obtained by detecting light absorption at 353 nm using dipping probe, in the broad concentration range from 0.3 to 60 mg/L with excellent linearity (R 2 = 0.9996). In situ measurements of adsorption and desorption kinetics on five kinds of MARs with different polarities were systematically carried out, showing that the adsorption process obeyed the pseudo-second-model. As well as, the system was proved to be highly accurate and reproducible. More importantly, this method enabled to study the initial stage of the adsorption process, starting from the time of the first second, which is the most important part in the adsorption kinetics, and this is impossible for traditional sampling methods. The successful application of FOS to in situ measurement of LPA not only contributes to fast, automatic, and real-time monitoring of LPA process but also enriches the research connotation of adsorption.

A Simple and Selective Fluorescent Sensor Chip for Indole-3-Butyric Acid in Mung Bean Sprouts Based on Molecularly Imprinted Polymer Coatings.

In this paper, we report the preparation of molecularly imprinted polymer coatings on quartz chips for selective solid-phase microextraction and fluorescence sensing of the auxin, indole-3-butyric acid. The multiple copolymerization method was used to prepare polymer coatings on silylated quartz chips. The polymer preparation conditions (e.g., the solvent, monomer, and cross-linker) were investigated systemically to enhance the binding performance of the imprinted coatings. Direct solid-phase fluorescence measurements on the chips facilitated monitoring changes in coating performance. The average binding capacity of an imprinted polymer coated chip was approximately 152.9 µg, which was higher than that of a non-imprinted polymer coated chip (60.8 µg); the imprinted coatings showed the highest binding to IBA among the structural analogues, indicating that the coatings possess high selectivity toward the template molecule. The developed method was used for the determination of the auxin in mung bean extraction, and the recovery was found to be in the range of 91.5% to 97.5%, with an RSD (n = 3) of less than 7.4%. Thus, the present study provides a simple method for fabricating a fluorescent sensor chip for selective analysis.

Lab-on-a-chip based biosensor for the real-time detection of aflatoxin.

Polymers were synthesized and utilized for aflatoxin detection coupled with a novel lab-on-a-chip biosensor: MiSens and high performance liquid chromatography (HPLC). Non-imprinted polymers (NIPs) were preferred to be designed and used due to the toxic nature of aflatoxin template and also to avoid difficult clean-up protocols. Towards an innovative miniaturized automated system, a novel biochip has been designed that consists of 6 working electrodes (1mm diameter) with shared reference and counter electrodes. The aflatoxin detection has been achieved by a competition immunoassay that has been performed using the new biochips and the automated MiSens electrochemical biosensor device. For the assay, aflatoxin antibody has been captured on the Protein A immobilized electrode. Subsequently the sample and the enzyme-aflatoxin conjugate mixture has been injected to the electrode surfaces. The final injection of the enzyme substrate results in an amperometric signal. The sensor assays for aflatoxin B1 (AFB1) in different matrices were also performed using enzyme link immunosorbent assay (ELISA) and HPLC for confirmation. High recovery was successfully achieved in spiked wheat samples using NIP coupled HPLC and NIP coupled MiSens biosensor [2ppb of aflatoxin was determined as 1.86ppb (93% recovery), 1.73ppb (86.5% recovery), 1.96ppb (98% recovery) and 1.88ppb (94.0% recovery) for immunoaffinity column (IAC)-HPLC, NIP-HPLC, Supel™ Tox SPE Cartridges (SUP)-HPLC and NIP-MiSens, respectively]. Aflatoxin detection in fig samples were also investigated with MiSens biosensor and the results were compared with HPLC method. The new biosensor allows real-time and on-site detection of AFB1 in foods with a rapid, sensitive, fully automated and miniaturized system and expected to have an immense economic impact for food industry.

Fast and sensitive detection of mycotoxins in wheat using microfluidics based Real-time Electrochemical Profiling.

The objective of the study has been the development of a new sensing platform, called Real-time Electrochemical Profiling (REP) that relies on real-time electrochemical immunoassay detection. The proposed REP platform consists of new electrode arrays that are easy to fabricate, has a small imprint allowing microfluidic system integration, enables multiplexed amperometric measurements and performs well in terms of electrochemical immunoassay detection as shown through the deoxynivalenol detection assays. The deoxynivalenol detection has been conducted according to an optimised REP assay protocol using deoxynivalenol standards at varying concentrations and a standard curve was obtained (y=-20.33ln(x)+124.06; R(2)=0.97) with a limit of detection of 6.25 ng/ml. As both ELISA and REP detection methods use horse radish peroxidase as the label and 3.3',5.5'-Tetramethylbenzidine as the substrate, the performance of the REP platform as an ELISA reader has also been investigated and a perfect correlation between the deoxynivalenol concentration and the current response was obtained (y=-14.56ln(x)+101.02; R(2)=0.99). The calibration curves of both assays have been compared to conventional ELISA tests for confirmation. After assay optimisation using toxin spiked buffer, the deoxynivalenol detection assay has also been performed to detect toxins in wheat grain.

On-line determination of 4-nitrophenol by combining molecularly imprinted solid-phase extraction and fiber-optic spectrophotometry.

In the present paper, we describe a new on-line SPE system where molecular imprinting, fiber-optic detection and flow injection analysis were combined for the first time. This new system has been applied for the on line detection of 4-nitrophenol (4-NP). Initially, molecularly imprinted polymers (MIP) have been prepared for the selective extraction of 4-NP using 4-vinylpyridine and ethylene glycol dimethacrylate as functional and cross-linking monomers, respectively. Selective extraction was achieved using the designed MIP with 97% of recovery on imprinted polymer and 10% on control polymer. The system provided a high degree of accuracy, with RSDs varying between 0.7 and 1.39%. In respect of accuracy, reproducibility, and rapidity, this system is comparable with HPLC. In short, the system allows simple, fast, and accurate analyte determination with the possibility of future automation.

Novel fluorescent membrane for metronidazole sensing prepared by covalent immobilization of a pyrenebutyric acid derivative.

In the present paper, we report the fabrication of a new sensing membrane for fluorescence detection of metronidazole (MNZ). Briefly, a pyrenebutyric acid derivative, 2-(methacryloyloxy) ethyl-4-(1-pyrenyl) butanoate (MPB) with a double bond, was synthesized and copolymerized with 2-hydroxyethylmethacrylate (HEMA) on the activated glass surface by thermal initiation in the presence of cross-linker. The sensor responds linearly to metronidazole in the concentration range of 1.23~35.48 mg.L(-1) in aqueous solution with a detection limit of 0.36 mg.L(-1). The lifetime is enhanced by covalently immobilizing the pyrenebutyric acid derivative on glass slide, which hinders leaching of the dye from the membrane. The sensor could be regenerated after use by washing in methanol (RSD = 2.42 %), and it shows sufficient stability, and selectivity. Interference of other pharmaceuticals on membrane performance is discussed. The developed membrane has been successfully applied for the direct determination of metronidazole in human serum sample without pretreatment.

Rational design of molecularly imprinted polymer: the choice of cross-linker.

The paper describes a rational approach for the selection of cross-linkers during the development of molecularly imprinted polymers (MIPs). As a model system for this research MIPs specific for the drug zidovudine (AZT) were designed and tested. Three cross-linkers trimethylolpropane trimethacrylate (TRIM), ethylene glycol dimethacrylate (EGDMA) and divinylbenzene (DVB) were studied. The analogue of zidovudine (AZT) ester (AZT-ES) was used as a dummy template. The imprinting factors for all of the polymers in the static adsorption experiments were calculated. The data on the AZT adsorption by control polymers (CP), which were prepared with different cross-linkers without a functional monomer, was also analyzed. DVB was found to be more inert towards zidovudine than EGDMA and TRIM, which was confirmed by both molecular modelling and adsorption experiments. It was demonstrated that DVB-based polymers had a higher imprinting factor (I = 1.85) compared with other tested cross-linked polymers. It was suggested that the selection of the cross-linker should be based on the strength of the interaction with the template: the cross-linker which displays lower binding of the template should be preferential because it generates MIPs with lower non-specific binding and a higher imprinting factor, and therefore specificity. Which cross-linker to use for the preparation of any particular MIP can be determined by analysis of the interactions between the cross-linker and template. This could be done either virtually using computational modelling or by template adsorption using a small library of polymers prepared using different cross-linkers.

Rational design and synthesis of water-compatible molecularly imprinted polymers for selective solid phase extraction of amiodarone.

Novel water-compatible molecularly imprinted polymers (MIPs) selective for amiodarone (AD) were designed via a new methodology which relies on screening library of non-imprinted polymers (NIPs). The NIP library consisted of eighteen cross-linked co-polymers synthesized from monomers commonly used in molecular imprinting. The binding capacity of each polymer in the library was analyzed in two different solvents. Binding in water was used to assess non-specific (hydrophobic) interactions and binding in an appropriate organic solvent was used to assess specific interactions. A good correlation was found between the screening tests and modeling of monomer-template interactions performed using computational approach. Additionally, analysis of template-monomer interactions was performed using UV-vis spectroscopy. As the result, 4-vinylpyridine (4-VP) was selected as the best monomer for developing MIP for AD. The 4-VP-based polymers demonstrated imprinting factor equal 3.9. The polymers performance in SPE was evaluated using AD and its structural analogues. The recovery of AD was as high as 96% when extracted from spiked phosphate buffer (pH 4.5) solution and 82.1% from spiked serum samples. The developed MIP shown as a material with specific binding to AD, comparing to its structural analogues, 1-(2-diethylaminoethoxy)-2,6-diiodo-4-nitrobenzene and lidocaine, which shown 9.9% and 25.4% of recovery from the buffer solution, correspondingly. We believe that the screening of NIP library could be proposed as an alternative to commonly used computational and combinatorial approaches.