Facile fabricate sandwich-structured molecularly imprinted dopamine polymer for simultaneously specific capture of Low-density lipoprotein and eliminate "bad cholesterol".

A simplified approach for preparation of sandwich type molecularly imprinted polymers (PPDA-MIPs) is proposed for simultaneously identify Low-density lipoprotein (LDL) and dispose "bad cholesterol". Porous polydopamine nanosphere (PPDA) is applied as a matrix for immobilization of LDL, and the imprinted layer is formed by dopamine acting as a functional monomer. Since imprinted cavities exhibit shape memory effects in terms of recognizing selectivity, the PPDA-MIPs exhibit excellent selectivity toward LDL and a substantial binding capacity of 550.3 µg mg-1. Meanwhile, six adsorption/desorption cycles later, the adsorption efficiency of 83.09 % is still achieved, indicating the adequate stability and reusability of PPDA-MIPs. Additionally, over 80 % of cholesterol is recovered, indicating the completeness of "bad cholesterol" removal in LDL. Lastly, as demonstrated by gel electrophoresis, PPDA-MIPs performed satisfactory behavior for the removal of LDL from the goat serum sample.

Integral methods for automatic quantification of fast-scan-cyclic-voltammetry detected neurotransmitters.

Modern techniques for estimating basal levels of electroactive neurotransmitters rely on the measurement of oxidative charges. This requires time integration of oxidation currents at certain intervals. Unfortunately, the selection of integration intervals relies on ad-hoc visual identification of peaks on the oxidation currents, which introduces sources of error and precludes the development of automated procedures necessary for analysis and quantification of neurotransmitter levels in large data sets. In an effort to improve charge quantification techniques, here we present novel methods for automatic selection of integration boundaries. Our results show that these methods allow quantification of oxidation reactions both in vitro and in vivo and of multiple analytes in vitro.

Addressing the instability issue of dopamine during microdialysis: the determination of dopamine, serotonin, methamphetamine and its metabolites in rat brain.

Dopamine is a catecholamine neurotransmitter that degrades rapidly in aqueous solutions; hence, its analysis following brain microdialysis is challenging. The aim of the current study was to develop and validate a new microdialysis coupled LC-MS/MS system with improved accuracy, precision, simplicity and turnaround time for dopamine, serotonin, methamphetamine, amphetamine, 4-hydroxymethamphetamine and 4-hydroxyamphetamine analysis in the brain. Dopamine degradation was studied with different stabilizing agents under different storage conditions. The modified microdialysis system was tested in vitro, and was optimized for best probe recovery, assessed by %gain. LC-MS/MS assay was developed and validated for the targeted compounds. Stabilizing agents (ascorbic acid, EDTA and acetic acid) as well as internal and cold standards were added on-line to the dialysate flow. Assay linearity range was 0.01-100 ng/mL, precision and accuracy passed criteria, and LOQ and LLOQ were 0.2 and 1.0 pg, respectively. The new microdialysis coupled LC-MS/MS system was used in Wistar rats striatum after 4 mg/kg subcutaneous methamphetamine. Methamphetamine rapidly distributed to rat striatum reaching an average ~200 ng/mL maximum, ~82.5 min post-dose. Amphetamine, followed by 4-hydroxymethamphetamine, was the most abundant metabolite. Dopamine was released following methamphetamine injection, while serotonin was not altered. In conclusion, we proposed and tested an innovative and simplified solution to improve stability, accuracy and turnover time to monitor unstable molecules, such as dopamine, by microdialysis.

Enhancement of fast scan cyclic voltammetry detection of dopamine with tryptophan-modified electrodes.

Fast scan cyclic voltammetry (FSCV) allows for real -time analysis of phasic neurotransmitter levels. Tryptophan (TRP) is an aromatic amino acid responsible for facilitating electron transfer kinetics in oxidoreductase enzymes. Previous work with TRP-modified electrodes showed increased sensitivity for cyclic voltammetry detection of dopamine (DA) when used with slower scan rates (0.05 V/s). Here, we outline an in vitro proof of concept for TRP-modified electrodes in FSCV detection of DA, and decreased sensitivity for ascorbic acid (AA). TRP-modified electrodes had a limit of detection (LOD) for DA of 2.480 ± 0.343 nM compared to 8.348 ± 0.405 nM for an uncoated electrode. Selectivity for DA/ascorbic acid (AA) was 1.107 ± 0.3643 for uncoated and 15.57 ± 4.184 for TRP-modified electrodes. Additionally, these TRP-modified electrodes demonstrated reproducibility when exposed to extended cycling. TRP-modified electrodes will provide an effective modification to increase sensitivity for DA.

N-Acetyldopamine derivatives from Periostracum Cicadae and their regulatory activities on Th1 and Th17 cell differentiation.

Bioassay-guided fractionation of a 90% ethanol extract of Periostracum Cicadae led to the isolation of two new N-acetyldopamine dimers (1a/1b) along with six known dimers (2a/2b, 3a/3b, and 4a/4b) and two monomers (5a/5b); compounds 2a/2b, 4a/4b and 5a/5b were newly isolated from this material. All compounds were isolated as enantiomeric mixtures and each enantiomer was successfully separated by chiral-phase HPLC. The structures including absolute configurations were confirmed by high-resolution electrospray ionization mass spectrometry (HR-ESIMS), 1D/2D nuclear magnetic resonance (NMR) spectroscopy, 1H iterative Full Spin Analysis (HiFSA), and electronic circular dichroism (ECD) spectroscopy. Subsequently, the bioactivities of these isolates were evaluated via CD4+ T cell differentiations, which are critical for immune responses and inflammation. The results revealed that compound 5b was observed to enhance the IFN-γ+ Th1 differentiation, which may have a potential for cancer immunotherapy.

In Matrix Derivatization Combined with LC-MS/MS Results in Ultrasensitive Quantification of Plasma Free Metanephrines and Catecholamines.

Plasma-free metanephrines and catecholamines are essential markers in the biochemical diagnosis and follow-up of neuroendocrine tumors and inborn errors of metabolism. However, their low circulating concentrations (in the nanomolar range) and poor fragmentation characteristics hinder facile simultaneous quantification by liquid chromatography and tandem mass spectrometry (LC-MS/MS). Here, we present a sensitive and simple matrix derivatization procedure using propionic anhydride that enables simultaneous quantification of unconjugated l-DOPA, catecholamines, and metanephrines in plasma by LC-MS/MS. Dilution of propionic anhydride 1:4 (v/v) in acetonitrile in combination with 50 µL of plasma resulted in the highest mass spectrometric response. In plasma, derivatization resulted in stable derivatives and increased sensitivity by a factor of 4-30 compared with a previous LC-MS/MS method for measuring plasma metanephrines in our laboratory. Furthermore, propionylation increased specificity, especially for 3-methoxytyramine, by preventing interference from antihypertensive medication (ß-blockers). The method was validated according to international guidelines and correlated with a hydrophilic interaction LC-MS/MS method for measuring plasma metanephrines (R2 > 0.99) and high-performance liquid chromatography with an electrochemical detection method for measuring plasma catecholamines (R2 > 0.85). Reference intervals for l-DOPA, catecholamines, and metanephrines in n = 115 healthy individuals were established. Our work shows that analytes in the subnanomolar range in plasma can be derivatized in situ without any preceding sample extraction. The developed method shows improved sensitivity and selectivity over existing methods and enables simultaneous quantification of several classes of amines.

Progress toward the development of a microchip electrophoresis separation-based sensor with electrochemical detection for on-line in vivo monitoring of catecholamines.

The development of a separation-based sensor for catecholamines based on microdialysis (MD) coupled to microchip electrophoresis (ME) with electrochemical (EC) detection is described. The device consists of a pyrolyzed photoresist film working electrode and a poly(dimethylsiloxane) microchip with a flow-gated sample injection interface. The chip was partially reversibly sealed to the glass substrate by selectively exposing only the top section of the chip to plasma. This partially reversible chip/electrode integration process not only allows the reuse of the working electrode but also greatly enhanced the reproducibility of electrode alignment with the separation channel. The developed MD-ME-EC system was then tested using l-DOPA, 3-O-MD, HVA, DOPAC, and dopamine standards, which were separated in less than 100 seconds using a background electrolyte consisting of 15 mM sodium phosphate (pH 7.4), 15 mM sodium dodecyl sulfate, and 2.5 mM boric acid. A potential of +1.0 V vs. Ag/AgCl was used for amperometric detection of the analytes. The device was evaluated for on-line monitoring of the conversion of l-DOPA to dopamine in vitro and for monitoring dopamine release in an anesthetized rat in vivo following high K+ stimulation. The system was able to detect stimulated dopamine release in vivo but not endogenous levels of dopamine.

Boronic acid-modified polyhedral oligomeric silsesquioxanes on polydopamine-coated magnetized graphene oxide for selective and high-capacity extraction of the catecholamines epinephrine, dopamine and isoprenaline.

Amino-functionalized polyhedral oligomeric silsesquioxanes (POSS-8NH2) were covalently bound to the surface of polydopamine-coated magnetized graphene oxide. It was then reacted with 4-formylphenylboronic acid to prepare a "cubic boronic acid"-bonded magnetic graphene oxide adsorbent. The new adsorbent exhibits better selectivity and much higher adsorption capacity for ortho-phenols over adsorbents where small boronic ligands are directly bound to the surface of the material. It is shown to enable selective and faster enrichment of the catecholamines epinephrine (EP), dopamine (DA) and isoprenaline (IP) with high selectivity over many potential interferents that can occur in urine. The analytes were then quantified by HPLC with fluorometric detection. Under optimal conditions, response is linear (R2 ≥ 0.9907), limits of detection are low (0.54-2.3 ng·mL-1), and reproducibility is acceptable (inter- and intra-day assay RSDs of≤10.9%). The method was successfully applied to the determination of endogenous EP and DA and exogenous IP in urine samples. Graphical abstractSchematic of boronic acid (BA)-modified polyhedral oligomeric silsesquioxanes (POSS) on polydopamine-coated magnetized graphene oxide (magGO). The material (magGO@POSS-BA) has good selectivity and higher adsorption capacity to ortho-phenols and can be applied to enrich the catecholamines in urine.

Comparison between electrochemical and photoelectrochemical detection of dopamine based on titania-ceria-graphene quantum dots nanocomposite.

In this study, titania-ceria-graphene quantum dot (TC-GQD) nanocomposite was synthesized by hydrothermal method for the first time. The prepared nanomaterials were characterized by XRD, FTIR dynamic light scattering (DLS), FESEM, HRTEM, and EDX spectroscopy along with elemental mapping. The synergistic effect of the nanocomposite components was studied by diffuse reflectance spectroscopy (DRS) and electrical conductivity meter. The results showed that band gap of TC-GQD nanocomposite was shifted to visible lights relative to its components (1.3 eV), and electrical conductivity of the sample was significant increased to 89.5 µS cm-1. After chemical and physical characterization, prepared new nanocomposites were used to design a new electrochemical (EC) and photoelectrochemical (PEC) dopamine (DA) sensors. In both EC and PEC methods effecting experimental parameters were optimized. Due to the synergic effect of the nanocomposite components, an outstanding photocurrent response was observed for DA based on PEC sensor. A linear calibration curve with a lower detection limit of 22 nM DA, and sensitivity of 13.8 mA/mM(DA), in a wider range of 0.3-750 µM DA, was obtained for TC-GQD/GCE electrode in PEC. While, the TC-GQD/GCE electrode detected DA in the range of 1-500 µM DA, with two linear calibration curve, detection limit of 0.22 µM DA, and sensitivity of 4.9 mA/mM(DA), in the EC. Observed results from EC and PEC sensors are presented and compared.

Mesoporous chitosan based conformable and resorbable biostrip for dopamine detection.

This work presents a chitosan based resorbable biostrip for label-free electrochemical detection of dopamine (DA).The biostrip consists of mesoporous-chitosan-graphene oxide (m-Chit-GO) composite-based sensing electrode and graphene-based interconnects. Obtained with particulate leaching, the m-chit-GO showed average pore size of 1µmwith slow (2 h) curing process. The response of DA on m-Chit-GO was investigated and compared with their bulk counterpart to study the effect of mesoporosity on voltammogram output signals. The voltammetric investigations were performed with three-electrode set-up using m-Chit-GO electrode as working electrode whereas Ag/AgCl and Graphene were used as a reference and counter electrodes, respectively. The quantitative analysis of concentration-dependent voltammetric peak-current enhancement revealed significantly higher response for m-Chit-GO (10pM) as compared to their bulk state (100 nM) on DA. The presented resorbable biostrip offers a limit of detection of 10pM and thereby shows great promise for detection of DA levels for early diagnosis of neurodegenerative diseases.

A selective colorimetric strategy for probing dopamine and levodopa through the mussel-inspired enhancement of Fe3O4 catalysis.

Mussel-inspired enhancement of Fe3O4 catalysis was discovered towards a highly selective and sensitive colorimetric strategy for the magnetic separation-based evaluation of dopamine and/or levodopa in urine, in which the specific interaction of bis-catechol-containing analytes and mesoporous Fe3O4 NPs would form highly stable complexes of bis-catechol-Fe coordination.

Design of Ni(OH)2 nanocages@MnO2 nanosheets core-shell architecture to jointly facilitate electrocatalytic dynamic for highly sensitive detection of dopamine.

In this work, Ni(OH)2 nanocages@MnO2 nanosheets core-shell architecture (Ni(OH)2 NCs@MnO2 NSs CSA) was successfully prepared through coordinated etching and precipitation (CEP) route followed by hydrothermal reaction, and then tested as sensitive electrode material for detection of dopamine (DA). The three dimensional (3D) hollow Ni(OH)2 core effectively prevented the aggregation of MnO2 NSs, leading to high utilization rate of MnO2 NSs. Meanwhile, the two dimensional (2D) MnO2 shell endowed Ni(OH)2 NCs with larger specific area and abundant diffusion channels, facilitating mass transport. Ni(OH)2 NCs@MnO2 NSs CSA modified glassy carbon electrode (GCE) exhibited two satisfying sensitivities of 467.1 and 1249.9 µA mM-1 cm-2 within the two linear ranges of 0.02-16.30 µM and 18.30-118.58 µM, respectively. Furthermore, Ni(OH)2 NCs@MnO2 NSs CSA/GCE presented low detection limit of 1.75 nM and short response time of 1.14 s. Overall, Ni(OH)2 NCs@MnO2 NSs/GCE looks promising for analytical sensing of DA thanks to its prominent electrocatalytic dynamic issued from the 3D hollow structure@2D nanosheets core-shell architecture.

Polymethacryloyl-L-Phenylalanine [PMAPA]-Based Monolithic Column for Capillary Electrochromatography.

The ability to detect catecholamines (CAs) and their metabolites is vital to understand the mechanism behind the neuronal diseases. Neurochemistry aims to provide an improved pharmacological, molecular and physiological understanding of complex brain chemistries by analytical techniques. Capillary electrophoresis (CE) is one such analytical technique that enables the study of various chemical species ranging from amino acids and peptides to natural products and drugs. CE can easily adapt the changes in research focus and in recent years remains an applicable technique for investigating neuroscience and single cell neurobiology. The prepared phenylalanine-based hydrophobic monolithic column, Polymethacryloyl-L-phenylalanine [PMAPA], was used as a stationary phase in capillary electrochromatography to separate CAs that are similar in size and shape to each other including dopamine (DA) and norepinephrine (NE) via hydrophobic interactions. Separation carried out in a short period of 17 min was performed with the electrophoretic mobility of 5.54 × 10-6 m2 V-1 s-1 and 7.60 × 10-6 m2 V-1 s-1 for DA and NE, respectively, at pH 7.0, 65% acetonitrile ratio with 100 mbar applied pressure by the developed hydrophobic monolithic column without needing any extra process such as imprinting or spacer arms to immobilize ligands used in separation.

Fluorescent sensor array for separation-free dopamine analogue discrimination via polyethyleneimine-mediated self-polymerization reaction.

The effective discrimination of dopamine (DA) analogues is an enduring challenge because of their very tiny structural differences, and thus a separation technique is generally required during the conventional analysis. In this study, a hyperbranched polyethyleneimine (hPEI)-based fluorescent sensor array has been constructed for the separation-free and effective differentiation of four DA analogues. The discrimination includes two steps: firstly, the formation of fluorescent polymer nanoparticles (FPNs) with diverse emission profiles via hPEI-mediated self-polymerization reaction of DA analogues and secondly, the linear discriminant analysis of fluorescence patterns of the formed FPNs for the differentiation of DA analogues. The hPEI-assisted self-polymerization reaction of DA analogues and substitution group mediated optical properties of the resulted FPNs enable an excellent discrimination of four DA analogues at a concentration of 1.0 µM when linear discriminant analysis and hierarchical cluster analysis are smartly combined. Additionally, binary, tertiary and even quaternary mixtures of analogues can also be well distinguished with the proposed sensor array. The practicability of this established sensor array is validated by a high accuracy (100%) evaluation of 88 blind samples containing a single analogue or a mixture of two, three or four analogues.

Ultra-selective fiber optic SPR platform for the sensing of dopamine in synthetic cerebrospinal fluid incorporating permselective nafion membrane and surface imprinted MWCNTs-PPy matrix.

Surface plasmon resonance (SPR) based dopamine sensor is realized using the state-of-art technique of molecular imprinting over an optical fiber substrate. Polypyrrole (PPy) is depicted as an effective polymer for the imprinting of dopamine through a green synthesis approach. Sensitivity of the probe is enhanced by the augmenting effect of surface imprinting of dopamine in polypyrrole over multiwalled carbon nanotubes (MWCNTs). To ensure the permselectivity of the probe towards dopamine molecules, a cation exchange polymer, nafion, is utilized as a membrane over imprinted sites to reduce the interference from anionic analytes like ascorbic acid and uric acid at physiological pH. The probe is characterized for a wide range of dopamine concentration from 0 to 10-5 M in artificial cerebrospinal fluid. Various probe parameters are varied to maximize the sensitivity of the sensor. The sensor possesses 18.9 pM as the limit of detection (LOD) which is lowest of those reported in the literature. The manifestation of sensing probe over an optical fiber along with the improved LOD makes the approach highly advantageous in terms of stability, repeatability, online remote monitoring, fast response, and miniaturization for its in vivo/in vitro applications in clinical sensing of dopamine.

3D hierarchical hollow hydrangea-like Fe3+@ɛ-MnO2 microspheres with excellent electrochemical performance for dopamine and hydrogen peroxide.

Construction a sensor to accurately detect dopamine (DA) and hydrogen peroxide (H2O2) is meaningful due to their close relation to the health of organisms. In this work, one-step hydrothermal method was employed to synthesize hierarchical hollow hydrangea-like Fe3+@ɛ-MnO2 microspheres constructed by interconnected nanosheets, and the growth mechanism of the microspheres (nFe/nMn = 0.6) was also investigated in detail. The material was used to construct an electrochemical sensor for DA and H2O2 detection with the linear range of 0.02-78 µmoL-1 and 0.000133-5.19 mmoL-1, respectively. The detection limit and sensitivity for DA and H2O2 are 5 and 50 nmoL-1 (S/N = 3), 7034.1 and 242.6 µA m(mol·L-1)-1 cm-2, respectively. Furthermore, the sensor was successfully applied to the detection of DA and H2O2 in serum and urine samples, indicating a potential value of this work in the pharmaceutical and environmental fields.

Wearable Fiber-Based Organic Electrochemical Transistors as a Platform for Highly Sensitive Dopamine Monitoring.

Fiber-based organic electrochemical transistors (FECTs) provide a new platform for the realization of an ultrafast and ultrasensitive biosensor, especially for the wearable dopamine (DA)-monitoring device. Here, we presented a fully filament-integrated fabric, it exhibited remarkable mechanical compatibility with the human body, and the minimum sensing unit was an organic electrochemical transistor (OECT) based on PVA- co-PE nanofibers (NFs) and polypyrrole (PPy) nanofiber network. The introduction of NFs notably increased the specific surface area and hydrophilicity of the PA6 filament, resulting in the formation of a large area of intertwined PPy nanofiber network. The electrical performance of PPy nanofiber network-modified fibers improved considerably. For the common FECTs, the typical on/off ratio was up to two orders of magnitude, and the temporal recovery time between on and off states was shortened to 0.34 s. Meanwhile, the device exhibited continuous cycling stability. In addition, the performances of FECT-based dopamine sensors depending on different gate electrodes have also been investigated. The PPy/NFs/PA6 filament-based dopamine sensor was more superior to the gold and platinum (Pt) wires, and the sensor presented long-term sensitivity with a detection region from 1 nM to 1 µM, rapid response time to a set of DA concentrations, remarkable selectivity in the presence of sodium chloride, uric acid, ascorbic acid and glucose, and superior reproducibility. Moreover, it could also be woven into the fabric product. The novel and wearable FECT device shows the potential to become the state-of-the-art DA-monitoring platform.

Nafion-Radical Hybrid Films on Carbon Nanotube Transistors for Monitoring Antipsychotic Drug Effects on Stimulated Dopamine Release.

We developed floating electrode-based carbon nanotube biosensors for the monitoring of antipsychotic drug effects on the dopamine release from PC12 cells under potassium stimulation. Here, carbon nanotube field-effect transistors with floating electrodes were functionalized with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•) radicals by Nafion films. This method allows us to build selective biosensors for dopamine detection with a detection limit down to 10 nM even in the presence of other neurotransmitters such as glutamate and acetylcholine, resulting from the selective interaction between ABTS• radicals and dopamine. The sensors were also utilized to monitor the real-time release of dopamine from PC12 cells upon the stimulation of high-concentrated potassium solutions. Significantly, the antipsychotic effects of pimozide on the dopamine release from potassium-stimulated PC12 cells could also be evaluated in a concentration-dependent manner by using the sensors. The quantitative and real-time evaluation capability of our strategy should provide a versatile tool for many biomedical studies and applications.

Affinity of Electrochemically Deposited Sol⁻Gel Silica Films towards Catecholamine Neurotransmitters.

Dopamine, norepinephrine, and epinephrine neurotransmitters can be detected by electrochemical oxidation in conventional electrodes. However, their similar chemical structure and electrochemical behavior makes a difficult selective analysis. In the present work, glassy carbon electrodes have been modified with silica layers, which were prepared by electroassisted deposition of sol⁻gel precursors. These layers were morphologically and compositionally characterized using different techniques, such as field emission scanning electron microscopy (FESEM), TEM, FTIR, or thermogravimetric analysis⁻mass spectrometry (TG-MS). The affinity of silica for neurotransmitters was evaluated, exclusively, by means of electrochemical methods. It was demonstrated that silica adsorbs dopamine, norepinephrine, and epinephrine, showing different interaction with silica pores. The adsorption process is dominated by a hydrogen bond between silanol groups located at the silica surface and the amine groups of neurotransmitters. Because of the different interaction with neurotransmitters, electrodes modified with silica films could be used in electrochemical sensors for the selective detection of such molecules.

Dopamine/2-Phenylethylamine Sensitivity of Ion-Selective Electrodes Based on Bifunctional-Symmetrical Boron Receptors.

Piperazine-based compounds bearing two phenylboronic acid or two benzoxaborole groups (PBPA and PBBB) were applied as dopamine receptors in polymeric membranes (PVC/DOS) of ion-selective electrodes. The potentiometric sensitivity and selectivity of the sensors towards dopamine were evaluated and compared with the results obtained for 2-phenylethylamine. Since the developed electrodes displayed strong interference from 2-phenylethylamine, single-molecule geometry optimizations were performed using the density functional theory (DFT) method in order to investigate the origin of dopamine/2-phenylethylamine selectivity. The results indicated that phenylboronic acid and benzoxaborole receptors bind dopamine mainly through the dative B⁻N bond (like 2-phenylethylamine) and the potentiometric selectivity is mainly governed by the higher lipophilicity of 2-phenylethylamine.