Hydrophilic interaction liquid chromatography based method for simultaneous determination of purines and their derivatives in food spices.

This work presents method for separation and quantification of adenine, guanine, xanthine, hypoxanthine, uric acid, and creatinine in food spices using hydrophilic interaction liquid chromatography with UV detection. Optimized conditions allowed separation with mobile phases containing acetonitrile and additives ammonium acetate (90:10, v/v, pH 6.1) or formate (90:10, v/v, pH 3.2). In food spices no uric acid was detected, creatinine (16 ± 2 µg g-1) was found only in instant dried yeast. The highest content of purines was determined in dried yeast (xanthine 110 ± 8 µg g-1, hypoxanthine 441 ± 24 µg g-1, adenine 84 ± 16 µg g-1, guanine 163 ± 12 µg g-1), high in curry, herbal pepper, and chicken seasoning, the lowest concentration was in black pepper (hypoxanthine 12 ± 2 µg g-1, adenine 27 ± 3 µg g-1). To best of our knowledge, no such complementary method and obtained data have been reported so far.

Exogenous application of xanthine and uric acid and nucleobase-ascorbate transporter MdNAT7 expression regulate salinity tolerance in apple.

BACKGROUND: Soil salinity is a critical threat to global agriculture. In plants, the accumulation of xanthine activates xanthine dehydrogenase (XDH), which catalyses the oxidation/conversion of xanthine to uric acid to remove excess reactive oxygen species (ROS). The nucleobase-ascorbate transporter (NAT) family is also known as the nucleobase-cation symporter (NCS) or AzgA-like family. NAT is known to transport xanthine and uric acid in plants. The expression of MdNAT is influenced by salinity stress in apple. RESULTS: In this study, we discovered that exogenous application of xanthine and uric acid enhanced the resistance of apple plants to salinity stress. In addition, MdNAT7 overexpression transgenic apple plants showed enhanced xanthine and uric acid concentrations and improved tolerance to salinity stress compared with nontransgenic plants, while opposite phenotypes were observed for MdNAT7 RNAi plants. These differences were probably due to the enhancement or impairment of ROS scavenging and ion homeostasis abilities. CONCLUSION: Our results demonstrate that xanthine and uric acid have potential uses in salt stress alleviation, and MdNAT7 can be utilized as a candidate gene to engineer resistance to salt stress in plants.

CuO Hollow Cubic Caves Wrapped with Biogenic N-Rich Graphitic C for Simultaneous Monitoring of Uric Acid and Xanthine.

Herein, we synthesized hollow cubic caves of CuO (HC) and wrapped it with N-rich graphitic C (NC), derived from a novel biogenic mixture composed of dopamine (DA) and purine. The synthesized NC wrapped HC (NC@HC) sensor shows enhanced electrocatalytic efficacy compared to unwrapped CuO with shapes including HC, sponge (SP), cabbage (CB), and solid icy cubes (SC). The shape and composition of synthesized materials were confirmed through field-emission scanning electron microscopy (FE-SEM), X-ray diffraction (XRD), Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS), whereas interfacial surface energy was calculated through contact angle measurement. The designed NC@HC sensor shows a remarkable response toward the simultaneous detection of uric acid (UA) and xanthine (Xn) with detection limits of 0.017 ± 0.001 (S/N of 3) and 0.004 ± 0.001 µM (S/N of 3), respectively. In addition, this platform was successfully applied to monitor UA from the gout patient serum. To the best of our knowledge, this is the first report on using such novel NC@HC materials for the simultaneous monitoring of UA and Xn.

Convenient synthesis of three-dimensional hierarchical CuS@Pd core-shell cauliflowers decorated on nitrogen-doped reduced graphene oxide for non-enzymatic electrochemical sensing of xanthine.

A novel hybrid with three-dimensional (3D) hierarchical CuS@Pd core-shell cauliflowers decorated on nitrogen-doped reduced graphene oxide (CuS@Pd/N-RGO) has been prepared by a facile wet-chemical route without utilizing any template molecules and surfactants. The characterization results reveal that the 3D flower-like structure of CuS "core" is composed of interconnecting nanoplates, which is conductive to the loading of Pd nanoparticles' "shell" and results in the robust interaction between the core and shell for the formation of CuS@Pd cauliflowers. Anchoring such appealing CuS@Pd cauliflowers on the two-dimensional N-RGO can efficaciously inhibit the aggregation of CuS@Pd cauliflowers and accelerate the kinetics of xanthine oxidation. Benefiting from the multi-functional properties and unique morphology, the sensor constructed by CuS@Pd/N-RGO exhibits excellent performance for non-enzymatic detection of xanthine including a wide detection range of 0.7-200.0 µM (0.94 V vs. SCE), a low detection limit of 28 nM (S/N = 3), high reproducibility (relative standard deviation (RSD) = 4.1%), and commendable stability (retained 90% of the initial electrochemical responses after storage for 30 days), which is amongst the best of various electrochemical sensors reported for xanthine assays till date. Reliable and satisfying recoveries (95-105%, RSD ≤ 4.1%) are achieved for xanthine detection in real samples. The inspiring results make the uniquely structural CuS@Pd/N-RGO greatly promising in non-enzymatic electrochemical sensing applications. Graphical abstract A high-performance non-enzymatic xanthine sensor has been constructed by the three-dimensional hierarchical CuS@Pd core-shell cauliflowers decorated on nitrogen-doped reduced graphene oxide.

Effect of allicin and its mechanism of action in purine removal in turbot.

Low-purine food is not only the focus of gout patients, but also the focus of contemporary green diet development. Fish are usually considered as high-purine foodstuff because of the high nutritional value and high purine content. Therefore, it is necessary to reduce purine content in fish to ensure that they are suitable for patients with hyperuricemia or gout. In this study, the effect of allicin on purine reduction in turbot during cooking was investigated using high-performance liquid chromatography, and the change of xanthine oxidase (XO) activity was also studied. Molecular docking analysis was further performed to elucidate the mechanism of purine reduction by allicin. The results revealed that in the step of soaking, allicin could reduce purine content in fish by slightly enhancing XO activity, promoting hypoxanthine transformation into xanthine. The removal of total purines in experimental and control group reached 70.45% and 57.20%, respectively. Moreover, allicin could change the thermal stability of xanthine by providing an acidic environment, resulting in the rapid decrease of xanthine and hypoxanthine levels by boiling. Thus, this study provides a simple method to decrease purine levels, suggesting a possibility that allicin can function as a purine remover in food.

Science and Healthy Meals in the World: Nutritional Epigenomics and Nutrigenetics of the Mediterranean Diet.

The Mediterranean Diet (MD), UNESCO Intangible Cultural Heritage of Humanity, has become a scientific topic of high interest due to its health benefits. The aim of this review is to pick up selected studies that report nutrigenomic or nutrigenetic data and recapitulate some of the biochemical/genomic/genetic aspects involved in the positive health effects of the MD. These include (i) the antioxidative potential of its constituents with protective effects against several diseases; (ii) the epigenetic and epigenomic effects exerted by food components, such as Indacaxanthin, Sulforaphane, and 3-Hydroxytyrosol among others, and their involvement in the modulation of miRNA expression; (iii) the existence of predisposing or protective human genotypes due to allelic diversities and the impact of the MD on disease risk. A part of the review is dedicated to the nutrigenomic effects of the main cooking methods used in the MD and also to a comparative analysis of the nutrigenomic properties of the MD and other diet regimens and non-MD-related aliments. Taking all the data into account, the traditional MD emerges as a diet with a high antioxidant and nutrigenomic modulation power, which is an example of the "Environment-Livings-Environment" relationship and an excellent patchwork of interconnected biological actions working toward human health.

A simple electrochemical approach to fabricate functionalized MWCNT-nanogold decorated PEDOT nanohybrid for simultaneous quantification of uric acid, xanthine and hypoxanthine.

Medical diagnostics and detection of food spoilage require estimation of hypoxanthine (HX), xanthine (XN), and uric acid (UA). A selective sensing platform has been proposed for simultaneous detection of all these species. Functionalized multi-walled carbon nanotube (fMWCNT) stabilized nanogold decorated PEDOT:TOS polymeric nanocomposite (Au-PEDOT-fMWCNT) was synthesized through rapid one-step electropolymerization to enhance conductivity and active surface area by several folds. Electrochemical activities of the proposed sensing platform were analyzed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS). Analyses through SEM, FESEM and TEM were performed to explore the surface morphology and elemental analysis of the polymeric nanohybrid was investigated by XPS, Raman, FTIR, XRD spectroscopy. Electro-catalysis of UA, XN and HX occurred at low oxidation potentials i.e. 0.082, 0.463 and 0.808 V, respectively in the optimized conditions. The uniquely designed simple, interference free Au-PEDOT-fMWCNT/GCE sensor exhibited high selectivity, good reproducibility, reusability (∼180 times) and stability (∼3 month) with excellent sensitivity of 1.73, 14.31 and 3.82 µA µM-1 cm-2 for UA, XN and HX, respectively. The sensor exhibited linear ranges of detection as 0.1-800, 0.05-175 and 0.1-150 µM with detection limits of 199.3, 24.1 and 90.5 nM for quantification of UA, XN and HX respectively. The performance of the proposed sensor was validated by addition of UA, XN and HX in human serum, urine and fish samples by comparing to those using HPLC. The results indicated good applicability of the proposed sensor for simultaneous detection of UA, XN, HX in real biological fluids.

Simple and Cost-effective "Turn-on" Fluorescence Sensor for the Determination of Xanthine.

A simple and selective 'turn-on' fluorescence sensor have been developed for the determination of xanthine (XA) based on glutathione (GSH) capped copper nanoclusters (CuNCs) as the fluorescent probe. The proposed sensor possess several advantages such as sensitivity, short analysis time and requires no sample pretreatment. The conditions for the performances of the sensor have been optimized and good linear relationship was obtained between concentration and relative fluorescence intensity in the concentration range 9.0[Formula: see text]10-3 M to 8.0[Formula: see text]10-5 M with a detection limit 6.0[Formula: see text]10-6 M. The mechanism behind the fluorescence enhancement may be ascribed to the binding of XA on the surface of GSH CuNCs. The sensor have been successfully applied to determine XA in spiked physiological samples.

Dual-Emission Reverse Change Ratio Photoluminescence Sensor Based on a Probe of Nitrogen-Doped Ti3C2 Quantum Dots@DAP to Detect H2O2 and Xanthine.

Titanium carbide quantum dots (Ti3C2 QDs) derived from two-dimensional (2D) Ti3C2Tx (MXene) are the rising-star material recently. Herein, nitrogen-doped Ti3C2 QDs (N-Ti3C2 QDs) were synthesized via a solvothermal method. The obtained N-Ti3C2 QDs exhibited excitation-dependent photoluminescence, antiphotobleaching, and dispersion stability. Furthermore, by combining the N-Ti3C2 QDs and DAP (2,3-diaminophenazine, the oxidative product of o-phenylenediamine) as a composite nanoprobe (N-Ti3C2 QDs@DAP), we developed a dual-emission reverse change ratiometric sensor to quantitatively monitor H2O2 based on photoinduced electron-transfer effects, where N-Ti3C2 QDs acted as the donor and DAP as the acceptor. On the basis of the xanthine converting into H2O2 through the catalysis of xanthine oxidase, the N-Ti3C2 QDs@DAP nanoprobe was also exploited for xanthine sensing. As a result, the proposed assay was demonstrated to be highly sensitive for H2O2 and xanthine with detection limits of 0.57 and 0.34 µM, respectively. In a word, we have investigated the application of N-Ti3C2 QDs in H2O2 and xanthine sensing and opened a new and exciting avenue for the N-Ti3C2 QDs in biosensing.

Hemin@carbon dot hybrid nanozymes with peroxidase mimicking properties for dual (colorimetric and fluorometric) sensing of hydrogen peroxide, glucose and xanthine.

The multifunctional hemin@carbon dot hybrid nanozymes (hemin@CD) with simultaneous peroxidase-like activity and fluorescence signalling property was prepared for the first time. Based on these properties, hemin@CD was applied to develop a dual-channel fluorescent probe for H2O2 and H2O2-based biocatalytic systems. By virtue of the peroxidase-like activity, hemin@CD can catalyze the oxidative coupling of 4-aminoantipyrine with phenol in the presence of H2O2 to form a pink-red quinoneimine dye with a maximum absorbance at 505 nm. Under the excitation wavelength of 480 nm, the green fluorescence of hemin@CD peaks at 540 nm and is quenched by the generated quinoneimine dye due to an inner filter effect, and also by H2O2 because of dynamic quenching. Thus, a colorimetric and fluorimetric dual-channel optical probe for H2O2 is obtained. Due to the glucose/xanthine transformations under formation of H2O2 by the relevant oxidase catalysis, the probe can be applied for detection of glucose and xanthine. The colorimetric detection limits for H2O2, glucose and xanthine are 0.11, 0.15, 0.11 µM, and the and fluorimetric detection limits are 0.15, 0.15, 0.12 µM, respectively. Graphical abstractSchematic representation of the colorimetric and fluorimetric dual probe for H2O2, glucose and xanthine based on the multifunctional emin@carbon dot) hybrid nanozymes with simultaneous peroxidase-like activity and fluorescence signalling property.

Recent progress in nanomaterial-based electrochemical and optical sensors for hypoxanthine and xanthine. A review.

This review (with 160 ref.) summarizes the progress that has been made in the methods for chemical or biochemical sensing of hypoxanthine and xanthine, which are produced as part of purine metabolism and are precursors of uric acid. An introduction discusses the importance of hypoxanthine and xanthine as analytes due to their significance in the clinical and food science, together with the conventional methods of analysis. A large section covers methods for the electrochemical hypoxanthine and xanthine sensing. It is divided into subsections according to the nanomaterials used including carbon nanomaterials, meal oxide nanoparticles, metal organic frameworks, conductive polymers, and bio-nanocomposites. A further large section covers optical methods for hypoxanthine and xanthine sensing, with subsections on nanomaterials including carbon nanomaterials, nanosheets, nanoclusters, nanoparticles, and their bio-nanocomposites. A concluding section summarizes the current status, addresses current challenges, and discusses future perspectives. Graphical abstractSchematic representation of the hypoxanthine and xanthine electrochemical and optical sensors incorporating various nanomaterials like graphene, carbon nanotubes (CNT), quantum dots (QD), nanoparticles and polymers, which are implemented in clinical and food analysis.

Triethylamine-controlled Cu-BTC frameworks for electrochemical sensing fish freshness.

The simultaneous determination of xanthine (XA) and hypoxanthine (HXA) has been proved to be a feasible approach for the assessment of fish freshness. In this study, copper(II) nitrate and 1,3,5-benzenetricarboxylic acid (H3BTC) were used as precursors to prepare various Cu-BTC frameworks with the addition of various amounts of triethylamine at room temperature. The characterization of X-ray diffraction, Fourier-transform infrared spectroscopy and Raman spectroscopy testified that the obtained materials are Cu-BTC frameworks. However, the amount of triethylamine had significant effects on the morphology, active response area and electron transfer ability of Cu-BTC frameworks. The oxidation behavior of XA and HXA demonstrated that the prepared Cu-BTC frameworks exhibited higher sensing activity, with greatly-enhanced oxidation signals. More importantly, the amount of triethylamine obviously affected the accumulation capacity and signal enhancement ability of Cu-BTCs toward XA and HXA, as confirmed from double potential step chronocoulometry. Based on the triethylamine-tuned signal amplification strategy of Cu-BTC frameworks, a highly-sensitive and simple electrochemical sensing system was developed for the assessment of fish freshness by simultaneous detection of XA and HXA. The developed sensing method was used in practical samples, and the results were validated by high-performance liquid chromatography.

Ultrafine Fe3C nanoparticles embedded in N-doped graphitic carbon sheets for simultaneous determination of ascorbic acid, dopamine, uric acid and xanthine.

A pyrolytic method is described for preparation of ultrafine Fe3C nanoparticles incorporated into N-doped graphitic carbon nanosheets (Fe3C@NGCSs). Iron phthalocyanine and graphitic carbon nitride (g-C3N4) are used as starting materials. The hybrid nanocomposite was placed on a glassy carbon electrode (GCE) and then applied to simultaneous determination of ascorbic acid (AA), dopamine (DA), uric acid (UA) and xanthine (XA). Figures of merits are as follows: for AA, the linear response range covers the 54.0-5491.0 µM range, the lower detection limit is 16.7 µM, and the best working voltage (vs. the saturated calomel electrode (SCE)) is 0.05 V. The respective data for DA are 1.2-120.8 µM, 0.34 µM and 0.19 V (vs. SCE). For UA, the respective data are 4.8-263.0 µM, 1.4 µM and 0.32 V (vs. SCE), and for XA the data are 4.8-361.0 µM, 1.5 µM and 0.71 V (vs. SCE). The method was successfully applied to their simultaneous determination in spiked serum samples. Graphical abstract Ultrafine Fe3C nanoparticles embedded in N-doped graphitic carbon sheets for simultaneous determination of ascorbic acid, dopamine, uric acid and xanthine.

Electrochemical xanthine detection by enzymatic method based on Ag doped ZnO nanoparticles by using polypyrrole.

A sensitive electrochemical detection of xanthine (X), which is an early biomarker of fish meat spoilage, was achieved by a novel biosensor developed via three main steps. The first step is the electropolymerization of a conducting polymer (pyrrole) onto the pencil graphite electrode (PGE). The second step is the entrapment of silver-doped zinc oxide nanoparticles (nano Ag-ZnO) onto PGE, which has already been doped with polypyrrole (PPy). The third step is the immobilization of the enzyme (xanthine oxidase) onto the modified electrode (nano Ag-ZnO/PPy/PGE) surface. The biosensor was characterized by scanning electron microscopy (SEM). The addition of Ag-doped ZnO nanoparticles into the conducting polymer structure played an important role in the performance of the biosensor by increasing the porous structure of the conducting polymer surface. The electrochemical behaviour of the biosensor was studied by electrochemical impedance spectroscopy (EIS) and chronoamperometry (CA). This enzyme biosensor showed the maximum response at pH 7.40 when +0.7 V was applied to reach 95% of steady-state current at ~3.2 s. The designed biosensor showed high selectivity with a sensitivity of 0.03 µA/mM and a low detection limit of 0.07 µM.

Bacterial Nucleobases Synergistically Induce Larval Settlement and Metamorphosis in the Invasive Mussel Mytilopsis sallei.

Marine bacterial biofilms have long been recognized as potential inducers of larval settlement and metamorphosis in marine invertebrates, but few chemical cues from bacteria have been identified. Here, we show that larval settlement and metamorphosis of an invasive fouling mussel, Mytilopsis sallei, could be induced by biofilms of bacteria isolated from its adult shells and other substrates from the natural environment. One of the strains isolated, Vibrio owensii MS-9, showed strong inducing activity which was attributed to the release of a mixture of nucleobases including uracil, thymine, xanthine, hypoxanthine, and guanine into seawater. In particular, the synergistic effect of hypoxanthine and guanine was sufficient for the inducing activity of V. owensii MS-9. The presence of two or three other nucleobases could enhance, to some extent, the activity of the mixture of hypoxanthine and guanine. Furthermore, we determined that bacteria producing higher concentrations of nucleobases were more likely to induce larval settlement and metamorphosis of M. sallei than were bacteria producing lower concentrations of nucleobases. The present study demonstrates that bacterial nucleobases play an important role in larval settlement and metamorphosis of marine invertebrates. This provides new insights into our understanding of the role of environmental bacteria in the colonization and aggregation of invasive fouling organisms and of the metabolites used as chemical mediators in cross-kingdom communication within aquatic systems.IMPORTANCE Invasive species are an increasingly serious problem globally. In aquatic ecosystems, invasive dreissenid mussels are well-known ecological and economic pests because they appear to effortlessly invade new environments and foul submerged structures with high-density aggregations. To efficiently control exotic mussel recruitment and colonization, the need to investigate the mechanisms of substrate selection for larval settlement and metamorphosis is apparent. Our work is one of very few to experimentally demonstrate that compounds produced by environmental bacteria play an important role in larval settlement and metamorphosis in marine invertebrates. Additionally, this study demonstrates that bacterial nucleobases can be used as chemical mediators in cross-kingdom communication within aquatic systems, which will enhance our understanding of how microbes induce larval settlement and metamorphosis of dreissenid mussels, and it furthermore may allow the development of new methods for application in antifouling.

Hereditary xanthinuria in a goat.

A 2-year-old mixed breed goat was presented for a 1-day history of anorexia and 1 week of weight loss. Serum biochemistry disclosed severe azotemia. Abdominal ultrasound examination showed decreased renal corticomedullary distinction, poor visualization of the renal pelves, and dilated ureters. On necropsy, the kidneys were small, the pelves were dilated, and the medulla was partially effaced by variably sized yellow nephroliths. Histologically, cortical and medullary tubules were distended by yellow-brown, multilayered crystals. Stone composition was 100% xanthine. Exonic sequencing of xanthine dehydrogenase (XDH) and molybdenum cofactor sulfurase (MOCOS) identified 2 putative pathogenic variants: a heterozygous XDH p.Leu128Pro variant and a homozygous MOCOS p.Asp303Gly variant. Variant frequencies were determined in 7 herd mates, 12 goats undergoing necropsy, and 443 goats from genome databases. The XDH variant was not present in any of these 462 goats. The MOCOS variant allele frequency was 0.03 overall, with 3 homozygotes detected. Hereditary xanthinuria is a recessive disorder in other species, but the XDH variant could be causal if the case goat is a compound heterozygote harboring a second variant in a regulatory region not analyzed or if the combination of the XDH and MOCOS variants together abolish XDH activity. Alternatively, the MOCOS variant alone could be causal despite the presence of other homozygotes, because hereditary xanthinuria in humans often is asymptomatic. Ours is the first report describing the clinical presentation and pathology associated with xanthine urolithiasis in a goat. The data support hereditary xanthinuria, but functional studies are needed to conclusively determine the causal variant(s).

Rhodamine B Chemiluminescence Improved by Mimetic AuCu Alloy Nanoclusters and Ultrasensitive Measurement of H2O2, Glucose and Xanthine.

The less stability and robustness, high-cost preparation and maintenance of natural enzymes, especially horseradish peroxidase (HRP), challenge researchers to introduce effective alternatives for their wide applications. Herein, the peroxidase-like activity of AuCu bimetal nanoclusters (AuCu NCs) was investigated in the rhodamine B-H2O2 chemiluminescence (CL) system. AuCu NCs could effectively catalyzed the CL reaction, and a high intensive emission intensity was obtained. A comprehensive study was implemented to examine the effects of different stabilizing ligands and Au/Cu ratios on the catalytic activity of obtained NCs. Comparison experiments were also expanded to include Au and Cu nanoparticles with different sizes, too. The results verified the superior catalytic activity of penicillamine-stabilized AuCu bimetal NCs containing 50% cu atoms. Finally, the analytical application of the introduced CL system showed great sensitivity for H2O2 detection, with a detection limit of 0.13 nM. Moreover, the developed CL method was able to measure glucose and xanthine over wide concentration ranges of 0.1 - 400 and 0.1 - 200 µM, respectively. The method also indicated satisfactory reliability, confirmed by standard reference materials.

Nitrogen compounds in Phacelia tanacetifolia Benth. honey: First time report on occurrence of (-)-5-epi-lithospermoside, uridine, adenine and xanthine in honey.

Lacy phacelia (Phacelia tanacetifolia Borkh.) honey composition was screened by UHPLC-DAD-QqTOF-MS. The targeted analysis revealed 6 major nitrogen compounds including aromatic amino acids (tyrosine, phenylalanine), purine derivatives (adenine, xanthine), nucleoside (uridine) and rare non-cyanogenic cyanoglucoside, (-)-5-epi-lithospermoside ((2Z)-2-[(4R,5R,6S)-4,5-dihydroxy-6-(ß-d-glucopyranosyl)oxycyclohex-2-en-1-ylidene]acetonitrile). Their identity was confirmed by different analytical tools: HRMS, co-chromatography with standard compound or comprehensive NMR experiments. All the compounds, except amino acids, were reported and determined in honey for the first time. The amount of the compounds was quantified in 16 unifloral phacelia samples: adenine (18.45 ±â€¯4.63 mg/kg), xanthine (10.53 ±â€¯2.98 mg/kg), uridine (42.84 ±â€¯9.26 mg/kg), tyrosine (14.66 ±â€¯10.22 mg/kg), (-)-5-epi-lithospermoside (70.61 ±â€¯31.37 mg/kg) and phenylalanine (20.41 ±â€¯11.99 mg/kg). The (-)-5-epi-lithospermoside content is significantly correlated with P. tanacetifolia pollen percentage (R2 = 0.5612, p < 0.001) and it is proposed as a potential marker of botanical origin for phacelia honey.

Amperometric biosensors based on carboxylated multiwalled carbon nanotubes-metal oxide nanoparticles-7,7,8,8-tetracyanoquinodimethane composite for the determination of xanthine.

A comparison of the analytical performances of two xanthine biosensors, based on the use of different metal oxide nanoparticles (MONPs: Co3O4 or Fe3O4)-modified carboxylated multiwalled carbon nanotubes (c-MWCNTs)-7,7',8,8'-tetracyanoquinodimethane (TCNQ)-chitosan (CHIT) composite, is discussed. Xanthine oxidase (XOD) enzyme was covalently attached to c-MWCNTs/MONPs/TCNQ/CHIT/GCE via N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxy succinimide (NHS) chemistry and the electrode surface was further modified with Nafion in order to minimize the effect of possible interfering substances. The results showed that analytical performance of the Fe3O4 based biosensor was better than the Co3O4 based biosensor. The linear working range, limit of detection and sensitivity were found to be 1.9×10-6-2.3×10-4M, 0.20µM (S/N=3), 25.07µAmM-1cm-2 for the Fe3O4 based biosensor and 1.9×10-6-1.2×10-4M, 0.36µM (S/N=3), 13.24µAmM-1cm-2 for the Co3O4 based biosensor, respectively. The purposed biosensors were applied in the determination of xanthine in coffee samples, and satisfactory results were obtained.

Inkjet-Printing Enzyme Inhibitory Assay Based on Determination of Ejection Volume.

An accurate, rapid, and cost-effective methodology for enzyme inhibitor assays is highly needed for large-scale screening to evaluate the efficacy of drugs at the molecular level. For the first time, we have developed an inkjet printing-based enzyme inhibition assay for the assessment of drug activity using a conventional inkjet printer composed of four cartridges. The methodology is based on the determination of the number of moles of the drug on the printed surface. The number of moles was quantified through the volume of substance ejected onto the printed surface. The volume ejected on the reaction spot was determined from the density of reagent ink solution and its weight loss after printing. A xanthine oxidase (XOD) inhibition assay was executed to quantitatively evaluate antioxidant activities of the drug based on the determination of the number of moles of the drug ejected by inkjet printing. The assay components of xanthine, nitro blue tetrazolium (NBT), superoxide dismutase (SOD)/drug, and XOD were printed systematically on A4 paper. A gradient range of the number of moles of SOD/drug printed on A4 paper could be successfully obtained. Because of the effect of enzyme activity inhibition, incrementally reduced NBT formazan colors appeared on the paper in a number-of-moles-dependent manner. The observed inhibitory mole (IM50) values of tested compounds exhibited a similar tendency in their activity order, compared to the IC50 values observed through absorption assay in well plates. Inkjet printing-based IM50 assessment consumed a significantly smaller reaction volume (by 2-3 orders of magnitude) and more rapid reaction time, compared to the well-plate-based absorption assay.