[Improved Indophenol Titration Method for Ascorbic Acid Using a Dropper and Electronic Balance: Enhanced Convenience and Efficiency].

The analysis of ascorbic acid using the 2,6-dichloroindophenol (DCIP) titration method is a well-established technique, but requires the skilled handling of a burette for accurate measurements. In the present study, we propose a modified DCIP titrimetric method that replaces the burette with a dropper and employs an electronic balance to measure the titrated amount by weight. The dropper used can be flexibly selected, allowing for a wide range of drop sizes, from large to very small. This modification offers several advantages, including lower skill requirements, a 43% reduction in the analysis time, a 50% decrease in sample/reagent consumption, and the ability to prepare DCIP standard solutions tailored to the concentration of ascorbic acid in the sample being analyzed. Our analysis of several food samples using this improved method showed that inherent issues of the DCIP method, such as determining the titration end point, could not be resolved. Nevertheless, the improved titration method remains more convenient and adaptable than the original approach using a burette, enabling quick and accurate analysis, especially for unskilled analysts.

Methodological considerations of the acetaminophen Detection Kit®: Involvement of molecular oxygen (O2) in an indophenol reaction.

The Acetaminophen Detection Kit® (Kanto Chemical Company Co. Inc., Tokyo, Japan) is a colorimetric test based on an indophenol reaction. The test involves three reactions: deproteination of the sample, hydrolysis of acetaminophen to yield p-aminophenol, and coupling p-aminophenol with a derivative of phenol in alkali conditions to form a blue-colored indophenol dye. The kit was devised to accomplish these three reactions with only two reagents, allowing the prompt diagnosis of acetaminophen overdose in emergency medicine. In the user instructions included with the kit and in reports introducing the kit, the chemical composition of the two reagents was not disclosed. Details about the composition can be found in the Safety Data Sheet from the manufacturer; however, there is little explanation about the principle (mechanism) of the coupling reaction. This lack of information appears to have hampered the use of this kit in forensic medicine. In this report, we conducted the coupling reaction by successively adding the two reagents to a p-aminophenol (intermediate molecule) solution with the reaction vessel open to the air and under an anaerobic condition. Development of the blue color was inhibited in the absence of air but gradually developed when the reaction vessel was opened to air. Thus, the coupling reaction is an oxidation-reduction (redox) reaction that requires molecular oxygen (O2) dissolved from the air to act as an oxidant. This finding corroborates statements in previous reports and will hopefully facilitate the use of the kit for forensic purposes.

Optical System Based on Nafion Membrane for the Detection of Ammonia in Blood Serum Samples.

The blood ammonia (NH3) level is one of the most important hepatic biomarkers for the diagnosis and monitoring of liver pathologies and infections. In this work, we developed an optimized optical biosensing method to extract and quantify the ammonia contained in complex-matrix samples emulating the blood serum. First, the approach was tested with solutions of phosphate-buffered saline (PBS) and ammonia chloride. Then, further trials were carried out with solutions of fetal bovine serum (FBS). The ammonia was extracted from the tested samples through a customized cell, and it was optically quantified by exploiting the indophenol reaction. The extraction cell included a cation-exchange membrane in Nafion, which was chemically pre-treated through cleaning procedures of sulfuric acid and hydrogen peroxide to keep a basic pH in the ammonia solution and to avoid contaminants in the membrane. From the NH3 solution, the indophenol reaction produced light-reactive indophenol dye molecules, which were used as colorimetric indicators. Through absorbance measurements of the indophenol dye solution at 670 nm wavelength, we were able to detect and quantify the ammonia level in the samples both with a spectrophotometer and a customized miniaturized read-out system, obtaining a detection limit of 0.029 µmol/mL.

Anti-Helicobacter pylori, anti-Inflammatory, and Antioxidant Activities of Trunk Bark of Alstonia boonei (Apocynaceae).

An ulcer is an erosion of the gastric mucosa that occurs following an imbalance between the aggression and protective factors and/or an infection with Helicobacter pylori (H. pylori). About 90-100% of duodenal ulcers and 70-80% of gastric ulcers are caused by H. pylori. The objective of this work was to evaluate in vitro the anti-H. pylori activity and then the anti-inflammatory and antioxidant properties of aqueous and methanol extracts of Alstonia boonei. The anti-H. pylori tests (CMI and antiureasic activity) were determined using the agar well diffusion method, the microbroth dilution method, and the measurement of ammonia production by the indophenol method; the anti-inflammatory properties were evaluated by inhibition of proteinases, denaturation of albumin, production of NO by macrophages, cell viability, and hemolysis of red blood cells by heat; then, the antioxidant properties were evaluated by the FRAP method (ferric reducing antioxidant power) and the DPPH (1,1-diphenyl-2-picrylhydrazyl) test. The results show that the best trapping of the DPPH radical was obtained with the methanol extract (EC50 = 8.91 µg/mL) compared to the aqueous extract (EC50 = 19.86 µg/mL). The methanol extract also showed greater iron-reducing activity than the aqueous extract and vitamin C. Furthermore, at the concentration of 200 µg/mL, the methanol extract showed a percentage (96.34%) strains of H. pylori higher than that of the aqueous extract (88.52%). The MIC90 of the methanol extract was lower than that of the aqueous extract. The methanol extract showed a higher percentage inhibition (85%) of urease than the aqueous extract (73%). The methanol extract at a concentration of 1000 µg/mL showed the greatest ability to inhibit proteinase activity, albumin denaturation, and red blood cell hemolysis; on the other hand, maximum cell viability and greater production of nitrite oxide by macrophages were obtained with the aqueous extract. Aqueous and methanol extracts of Alstonia boonei possess anti-H. pylori which would probably be linked to their antioxidant and anti-inflammatory properties.

Sustainable removal of nitrite waste to value-added ammonia on Cu@Cu2O core-shell nanostructures by pulsed laser technique.

The biochemical reduction of nitrite (NO2-) ions to ammonia (NH3) requires six electrons and is catalyzed by the cytochrome c NO2- reductase enzyme. This biological reaction inspired scientists to explore the reduction of nitrogen oxyanions, such as nitrate (NO3-) and NO2- in wastewater, to produce the more valuable NH3 product. It is widely known that copper (Cu)-based nanoparticles (NPs) are selective for the NO3- reduction reaction (NO3-RR), but the NO2-RR has not been well explored. Therefore, we attempted to address the electrocatalytic conversion of NO2- to NH3 using Cu@Cu2O core-shell NPs to simultaneously treat wastewater by removing NO2- and producing valuable NH3. The Cu@Cu2O core-shell NPs were constructed using the pulsed laser ablation of Cu sheet metal in water. The core-shell nanostructure of these particles was confirmed by various characterization techniques. Subsequently, the removal of NO2- and the ammonium (NH4+)-N yield rate were estimated using the Griess and indophenol blue methods, respectively. Impressively, the Cu@Cu2O core-shell NPs exhibited outstanding NO2-RR activity, demonstrating a maximum NO2- removal efficiency of approximately 94% and a high NH4+-N yield rate of approximately 0.03 mmol h-1.cm-2 at -1.6 V vs. a silver/silver chloride reference electrode under optimal conditions. The proposed NO2-RR mechanism revealed that the (111) facet of Cu favors the selective conversion of NO2- to NH3 via a six-electron transfer. This investigation may offer a new insight for the rational design and detailed mechanistic understanding of electrocatalyst architecture for the effective conversion of NO2- to NH4+.

Combined liquid phase microextraction and fiber-optics-based cuvetteless micro-spectrophotometry for sensitive determination of ammonia in water and food samples by the indophenol reaction.

The Berthelot reaction for ammonia is revisited with the aim of miniaturization and addressing interferences as encountered with food and water samples. Headspace single drop microextraction of ammonia in phosphoric acid served to attain selectivity in complex matrices, and liquid-liquid microextraction of red or blue indophenol species into 1-octanol-isooctane (60:40, v/v) resulted into high sensitivity. Fiber-optics-based cuvetteless micro-spectrophotometry has been used for colorimetric determination on microliter volumes of extract. The linear dynamic range, limit of detection and enrichment factor have been found to be 0.2-3 mg kg-1, 0.14 mg kg-1 and 38, respectively, measuring red species for milk, cheese and beer (4.9-5.5% error; 4.8-6.3% RSD; n = 5); and 5-400 µg L-1, 0.4 µg L-1 and 137, respectively, measuring blue species for water samples (3.3-5.7% error; 3.6-6.8% RSD; n = 5). A plausible reaction scheme has been proposed for nitroprusside catalysis in indophenol reaction.

Development of an Integrated Syringe-Pump-Based Environmental-Water Analyzer ( iSEA) and Application of It for Fully Automated Real-Time Determination of Ammonium in Fresh Water.

The development of a multipurpose integrated syringe-pump-based environmental-water analyzer ( iSEA) and its application for spectrophotometric determination of ammonium is presented. The iSEA consists of a mini-syringe pump equipped with a selection valve and laboratory-programmed software written by LabVIEW. The chemistry is based on a modified indophenol method using o-phenylphenol. The effect of reagent concentrations and sample temperatures was evaluated. This fully automated analyzer had a detection limit of 0.12 µM with sample throughput of 12 h-1. Relative standard deviations at different concentrations (0-20 µM) were 0.23-3.36% ( n = 3-11) and 1.0% ( n = 144, in 24 h of continuous measurement, ∼5 µM). Calibration curves were linear ( R2 = 0.9998) over the range of 0-20 and 0-70 µM for the detection at 700 and 600 nm, respectively. The iSEA was applied in continuous real-time monitoring of ammonium variations in a river for 24 h and 14 days. A total of 1802 samples were measured, and only 0.4% was outlier data (≥3 sigma residuals). Measurements of reference materials and different aqueous samples ( n = 26) showed no significant difference between results obtained by reference and present methods. The system is compact (18 cm × 22 cm × 24 cm), portable (4.8 kg), and robust (high-resolution real-time monitoring in harsh environments) and consumes a small amount of chemicals (20-30 µL/run) and sample/standards (2.9 mL/run).

An LC-MS Approach to Quantitative Measurement of Ammonia Isotopologues.

Ammonia is a fundamental aspect of metabolism spanning all of phylogeny. Metabolomics, including metabolic tracing studies, are an integral part of elucidating the role of ammonia in these systems. However, current methods for measurement of ammonia are spectrophotometric, and cannot distinguish isotopologues of ammonia, significantly limiting metabolic tracing studies. Here, we describe a novel LC-MS-based method that quantitatively assesses both 14N-and 15N-isotopologues of ammonia in polar metabolite extracts. This assay (1) quantitatively measures the concentration of ammonia in polar metabolite isolates used for metabolomic studies, and (2) accurately determines the percent isotope abundance of 15N-ammonia in a cell lysate for 15N-isotope tracing studies. We apply this assay to quantitatively measure glutamine-derived ammonia in lung cancer cell lines with differential expression of glutaminase.

Lipopolysaccharide aggravated DOI-induced Tourette syndrome: elaboration for recurrence of Tourette syndrome.

Tourette syndrome (TS) is a neurological disorder characterized by highest familial recurrence rate among neuropsychiatric diseases with complicated inheritance. Recurrence of Tourette syndrome was frequently observed in clinical. Unexpectedly, the mechanism of recurrence of Tourette syndrome was failure to elucidate. Here, we first shown that lipopolysaccharide(LPS) may played an important role in the recurrence of Tourette syndrome. The TS model in rats was induced by DOI (the selective 5-HT2A/2C agonist 1-(2, 5-dimethoxy-4-iodophenyl) -2- aminopropane). The rats were randomly divided into 4 groups:(1)Control;(2) Control + LPS; (2)TS; (3)TS + LPS. The results demonstrated that the LPS treatment significantly increased stereotypic score and autonomic activity. LPS treatment also significantly increased inflammatory cytokines such as interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in serum and striatum. Also, highly expressed TLR4, MyD88, P-NF-κBp65, P-IκBα in TS rats were increased respectively by LPS treatment as indicted in western blot analysis and immunohistochemistry analysis. Thus, it was supposed that lipopolysaccharide(LPS) may played an important role in the recurrence of Tourette syndrome and its mechanism was related to TLR/NF-κB pathway.

Antibody-Free Colorimetric Detection of Total Aflatoxins in Rice Based on a Simple Two-Step Chromogenic Reaction.

The prevalently used immunoassays for fast screening of aftatoxins (AFs) usually cannot meet the requirement for simultaneous determination of total AFs (aflatoxin B1 + aflatoxin B2 + aflatoxin G1 + aflatoxin G2) due to the deficiency of highly group-specific antibodies. This paper describes a two-step chromogenic reaction based method to quantitatively detect total AFs in rice using colorimetric measurement without antibody. In the method, colorless AFs transform into green-colored indophenol products through the reaction with sodium hydroxide and 2,6-dibromoquinone-4-chloroimide (DBQC) successively, allowing selectively determining total AFs up to 3.9 µg/kg over other competitive mycotoxins under optimal conditions by a UV-vis spectrophotometer. In addition, the colorimetric measurement results of the rice samples agree well with that of a standard HPLC method, demonstrating the good reliability and applicability of the method. Uniquely, the method has potential for on-site detection of total AFs in rice when using a nylon membrane-based device.

Liquid waveguide spectrophotometric measurement of nanomolar ammonium in seawater based on the indophenol reaction with o-phenylphenol (OPP).

We describe a highly sensitive colorimetric method for the determination of nanomolar concentrations of ammonium in seawater based on the indophenol reaction with o-phenylphenol [(1,1'-biphenyl)-2-ol, abbreviated as OPP]. OPP is available as non-toxic, stable flaky crystals with no caustic odor and has some advantages over phenol in practical use. The method was established by using a gas-segmented continuous flow analyzer equipped with two types of long path liquid waveguide capillary cell, LWCCs (100 cm and 200 cm) and an UltraPath (200 cm), which have inner diameters of 0.55 mm and 2 mm, respectively. The reagent concentrations, flow rates of the pumping tubes, and reaction path and temperature were determined on the basis of a manual indophenol blue method with OPP (Kanda, Water Res. 29 (1995) 2746-2750). The sample mixed with reagents that form indophenol blue dye was measured at 670 nm. Aged subtropical surface water was used as a blank, a matrix of standards, and the carrier. The detection limits of the analytical systems with a 100 cm LWCC, a 200 cm LWCC, and a 200 cm UltraPath were 6, 4, and 4 nM, respectively. These systems had high precision (<4% at 100 nM) and a linear dynamic range up to 200 nM. Non-linear baseline drift did not occur when using the UltraPath system. This is due to the elimination of cell clogging because of the larger inner diameter of the UltraPath compared to the LWCCs. The UltraPath system is thus more suitable for long-term measurements compared with the LWCC systems. The results of the proposed sensitive colorimetry and a conventional colorimetry for the determination of seawater samples showed no significant difference. The proposed analytical systems were applied to underway surface monitoring and vertical observation in the oligotrophic South Pacific.

Simple and inexpensive quantification of ammonia in whole blood.

Quantification of ammonia in whole blood has applications in the diagnosis and management of many hepatic diseases, including cirrhosis and rare urea cycle disorders, amounting to more than 5 million patients in the United States. Current techniques for ammonia measurement suffer from limited range, poor resolution, false positives or large, complex sensor set-ups. Here we demonstrate a technique utilizing inexpensive reagents and simple methods for quantifying ammonia in 100 µL of whole blood. The sensor comprises a modified form of the indophenol reaction, which resists sources of destructive interference in blood, in conjunction with a cation-exchange membrane. The presented sensing scheme is selective against other amine containing molecules such as amino acids and has a shelf life of at least 50 days. Additionally, the resulting system has high sensitivity and allows for the accurate reliable quantification of ammonia in whole human blood samples at a minimum range of 25 to 500 µM, which is clinically for rare hyperammonemic disorders and liver disease. Furthermore, concentrations of 50 and 100 µM ammonia could be reliably discerned with p = 0.0001.

An enzymatic assay for high-throughput screening of cytidine-producing microbial strains.

Cytidine is an industrially useful precursor for the production of antiviral compounds and a variety of industrial compounds. Interest in the microbial production of cytidine has grown recently and high-throughput screening of cytidine over-producers is an important approach in large-scale industrial production using microorganisms. An enzymatic assay for cytidine was developed combining cytidine deaminase (CDA) and indophenol method. CDA catalyzes the cleavage of cytidine to uridine and NH3, the latter of which can be accurately determined using the indophenol method. The assay was performed in 96-well plates and had a linear detection range of cytidine of 0.058-10 mM. This assay was used to determine the amount of cytidine in fermentation flasks and the results were compared with that of High Perfomance Liquid Chromatography (HPLC) method. The detection range of the CDA method is not as wide as that of the HPLC, furthermore the correlation factor of CDA method is not as high as that of HPLC. However, it was suitable for the detection of large numbers of crude samples and was applied to high-throughput screening for high cytidine-producing strains using 96-well deep-hole culture plates. This assay was proved to be simple, accurate, specific and suitable for cytidine detection and high-throughput screening of cytidine-producing strains in large numbers of samples (96 well or more).

A rapid enzymatic assay for high-throughput screening of adenosine-producing strains.

Adenosine is a major local regulator of tissue function and industrially useful as precursor for the production of medicinal nucleoside substances. High-throughput screening of adenosine overproducers is important for industrial microorganism breeding. An enzymatic assay of adenosine was developed by combined adenosine deaminase (ADA) with indophenol method. The ADA catalyzes the cleavage of adenosine to inosine and NH3 , the latter can be accurately determined by indophenol method. The assay system was optimized to deliver a good performance and could tolerate the addition of inorganic salts and many nutrition components to the assay mixtures. Adenosine could be accurately determined by this assay using 96-well microplates. Spike and recovery tests showed that this assay can accurately and reproducibly determine increases in adenosine in fermentation broth without any pretreatment to remove proteins and potentially interfering low-molecular-weight molecules. This assay was also applied to high-throughput screening for high adenosine-producing strains. The high selectivity and accuracy of the ADA assay provides rapid and high-throughput analysis of adenosine in large numbers of samples.

Quantitative determination of glycine in aqueous solution using glutamate dehydrogenase-immobilized glyoxal agarose beads.

In this study, an enzymatic procedure for the determination of glycine (Gly) was developed by using a column containing immobilized glutamate dehydrogenase (GDH) on glyoxal agarose beads. Ammonia is produced from the enzymatic reactions between Gly and GDH with NAD(+) in phosphate buffer medium. The indophenol blue method was used for ammonia detection based on the spectrophotometric measurements of blue-colored product absorbing at 640 nm. The calibration graph is linear in the range of 0.1-10 mM of Gly concentrations. The effect of pH, temperature, and time interval was studied to find column stability, and also the interference effects of other amino acids was investigated. The interaction between GDH and glyoxal agarose beads was analyzed by Fourier transform infrared (FTIR) spectroscopy. The morphology of the immobilized and non-immobilized agarose beads were characterized by atomic force microscopy (AFM).

Interferencia de la bilirrubina en la medición del paracetamol por el método del indofenol en el analizador Dimension (R) EXL(TM) / Bilirubin interference in the measurement of paracetamol by the indophenol method on the Dimension (R) EXL(TM)

Introducción. Los niveles de paracetamol se miden habitualmente en los laboratorios de urgencias ante la evidencia o sospecha de una intoxicación para predecir la posible hepatotoxicidad aguda por este fármaco. Sin embargo varios estudios han reportado un falso incremento en la concentración de paracetamol medida por métodos enzimáticos en plasmas ictéricos, circunstancia que ocurre frecuentemente en casos de fracaso hepático. Material y métodos. Para evaluar la influencia de la bilirrubina en la medición de paracetamol en el analizador Dimension(R) EXL(TM) (Siemens Healthcare Diagnostics Inc.(R)) se preparó una solución primaria de interferente a una concentración de 800 mg/dl (13.680 μmol/l) que se añadió a una solución de control comercial con altos niveles de paracetamol hasta obtener una mezcla con una concentración de bilirrubina de 40 mg/dl (684 μmol/l) realizando la medición de paracetamol en esta mezcla y en una de referencia. Comprobada la existencia de interferencia se procedió a cuantificarla diluyendo la solución primaria hasta obtener mezclas con 20; 10; 5; 2,5; 1,25 y 0,625 mg/dl de bilirrubina (342; 171; 85,5; 42,75; 21,37 y 19,69 μmol/l). Se obtuvo el interferograma representando la desviación de la medida frente a la mezcla de referencia frente a las distintas concentraciones de interferente. Resultados. Las distribuciones fueron gaussianas, por lo que se pudo aplicar la t de Student para el cálculo de las significaciones estadísticas. Se demostró la existencia de una interferencia significativa negativa a partir de 5 mg/dl (85,5 μmol/l) de bilirrubina, lo que muestra que las mediciones de paracetamol deben interpretarse con cautela en casos de hiperbilirrubinemia (AU)

Background. Blood levels of acetaminophen (paracetamol) are measured routinely in large acetaminophen overdoses or when poisoning is suspected, in order to predict the likelihood of hepatotoxicity. However some cases of spurious acetaminophen levels have been reported in jaundiced patients. Material and methods. The interference of bilirubin in acetaminophen measurements was studied on the Dimension(R) EXL(T)M by preparing a primary mixture of bilirubin at 800 mg/dL, which was spiked to a quality control solution with high acetaminophen concentration in order to get a bilirrubin concentration of 40 mg/dl on the mixture. Acetaminophen concentration was measured in the mixture with bilirubin and in reference solution without bilirubin. Interference was assessed and quantified by diluting the bilirubin solution to achieve final concentrations of 20, 10, 5, 2.5, 1.25 and 0.625 mg/dl (342, 171, 85.5, 42.75, 21.37 and 19.69 μmol/l). A graph of the interference was obtained showing the bias of acetaminophen concentrations with reference solution vs. increasing bilirubin concentrations. Results. Acetaminophen concentrations in both solutions followed Gaussian distributions. The t test showed significant negative interference at bilirubin concentrations higher to 5 mg/dl (85.5 μmol/l), which means that blood acetaminophen concentrations must be interpreted with caution in cases of hyperbilirubinemia (AU)

Disruption of thalamocortical activity in schizophrenia models: relevance to antipsychotic drug action.

Non-competitive NMDA receptor antagonists are widely used as pharmacological models of schizophrenia due to their ability to evoke the symptoms of the illness. Likewise, serotonergic hallucinogens, acting on 5-HT(2A) receptors, induce perceptual and behavioural alterations possibly related to psychotic symptoms. The neurobiological basis of these alterations is not fully elucidated. Data obtained in recent years revealed that the NMDA receptor antagonist phencyclidine (PCP) and the serotonergic hallucinogen 1-(2,5-dimethoxy-4-iodophenyl-2-aminopropane; DOI) produce a series of common actions in rodent prefrontal cortex (PFC) that may underlie psychotomimetic effects. Hence, both agents markedly disrupt PFC function by altering pyramidal neuron discharge (with an overall increase) and reducing the power of low frequency cortical oscillations (LFCO; < 4 Hz). In parallel, PCP increased c-fos expression in excitatory neurons of various cortical areas, the thalamus and other subcortical structures, such as the amygdala. Electrophysiological studies revealed that PCP altered similarly the function of the centromedial and mediodorsal nuclei of the thalamus, reciprocally connected with PFC, suggesting that its psychotomimetic properties are mediated by an alteration of thalamocortical activity (the effect of DOI was not examined in the thalamus). Interestingly, the observed effects were prevented or reversed by the antipsychotic drugs clozapine and haloperidol, supporting that the disruption of PFC activity is intimately related to the psychotomimetic activity of these agents. Overall, the present experimental model can be successfully used to elucidate the neurobiological basis of schizophrenia symptoms and to examine the potential antipsychotic activity of new drugs in development.

The effect of the 5-HT2A/2C receptor agonist DOI on micturition in rats with chronic spinal cord injury.

PURPOSE: We examined the effects of the 5-HT2A/2C receptor agonist DOI on micturition in chronic spinal cord injured rats. MATERIALS AND METHODS: Female Sprague-Dawley® rats were used. Spinal cord injury was produced by transection at the T10 level. A cystometric study was performed 8 to 12 weeks after transection. Cystometrograms were done using urethane anesthesia in all rats. The selective 5-HT2A antagonist ketanserin was administered after each DOI dose-response curve. All drugs were administered intravenously. RESULTS: Compared to controls, spinal cord injured rats had higher bladder capacity and post-void residual urine volume, and lower voiding efficiency. In spinal cord injured rats DOI (0.01 to 0.3 mg/kg) induced significant dose dependent increases in micturition volume and decreases in residual volume, resulting in increased voiding efficiency. Cystometrogram measurements showed a dose dependent increase in high frequency oscillation activity, evident as an increased number of small oscillation per voiding. This correlated with the improved voiding efficiency. Ketanserin (0.1 mg/kg) partially or completely reversed the DOI induced changes. CONCLUSIONS: High frequency oscillation seems to reflect external urethral sphincter burst activity during voiding. Bladder voiding efficiency and high frequency oscillation activity were decreased in spinal cord injured rats. High frequency oscillation activity can be enhanced by 5-HT2A receptor agonism, improving voiding efficiency. To our knowledge it remains to be studied whether these results may have implications for the future treatment of voiding dysfunction in patients with spinal cord injury.

Liquid microjunction surface sampling probe fluid dynamics: characterization and application of an analyte plug formation operational mode.

The recently discovered sample plug formation and injection operational mode of a continuous flow, coaxial tube geometry, liquid microjunction surface sampling probe (LMJ-SSP) was further characterized and applied for concentration and mixing of analyte extracted from multiple areas on a surface and for nanoliter-scale chemical reactions of sampled material. A transparent LMJ-SSP was constructed and colored analytes were used so that the surface sampling process, plug formation, and the chemical reactions could be visually monitored at the sampling end of the probe before being analyzed by mass spectrometry of the injected sample plug. Injection plug peak widths were consistent for plug hold times as long as the 8 min maximum attempted (RSD below 1.5%). Furthermore, integrated injection peak signals were not significantly different for the range of hold times investigated. The ability to extract and completely mix individual samples within a fixed volume at the sampling end of the probe was demonstrated and a linear mass spectral response to the number of equivalent analyte spots sampled was observed. Using the color and mass changing chemical reduction of the redox dye 2,6-dichlorophenol-indophenol with ascorbic acid, the ability to sample, concentrate, and efficiently run reactions within the same plug volume within the probe was demonstrated.