A simple,rapid, and accurate fluorometric analysis of epinephrine in local anesthetic solutions

A method is presented for the fluorometric analysis of epinephrine contained in local anesthetic solutions. The method is simple, rapid, and accurate, and does not require separation techniques.     

Electrocatalytic Application of an Overoxidized Dopamine Film Prepared on a Gold Electrode Surface to Selective Epinephrine Sensing

A dopamine polymer film was prepared ex situ on a bare gold template from a 10 mM dopamine solution in phosphate buffer, pH 7 followed by an overoxidation in 500 mM NaOH. The modified electrode was used for quantitative determination of epinephrine. A linear relationship between epinephrine concentration and current response was obtained in the range between 2 μM and 800 μM with the detection limit of 0.3 μM. The results have shown that using the overoxidized dopamine film it is possible to perform electrochemical analysis of epinephrine without interference of ascorbic and uric acids.     

A rapid and simple method for determination of halothane, isoflurane and sevoflurane in blood using gas chromatography

We have developed a technique to determine the concentration of volatile anesthetics (halothane, isoflurane and sevoflurane) in blood that is a modification of a method used for volatile anesthetics in Krebs solution. Methylene chloride was the internal standard and chloroform was used to extract the volatile anesthetic from blood. The congealed blood proteins were separated from the chloroform solvent (containing anesthetic) using a two-compartment vial that filtered out the proteinaceous material during centrifuging. Recovery averaged 102%. Linearity was excellent (r = 0.992-0.999) in the 50-600, 50-300 and 50-300 microg/mL range for halothane, isoflurane and sevoflurane, respectively. Intra-day and inter-day precisions were likewise excellent, with relative standard deviations <5.3 and <7.1%, respectively. Accuracy ranged from 0.8 to 9.5% of the estimated theoretical value. Extracted anesthetic in chloroform solvent was stable over 4-5 days, with <3% variability. The time from obtaining the blood sample to determination of the concentration from the chromatographic peak was 15 min or less.     

Flow injection chemiluminescence determination of epinephrine using epinephrine-imprinted polymer as recognition material

A flow injection chemiluminescence (CL) method using epinephrine-imprinted polymer as recognition material was investigated for the determination of epinephrine. The analyte, epinephrine, was selectively and temporarily adsorbed on the epinephrine-imprinted polymer, which was packed into a flow-cell placed in front of the window of a photomultiplier tube. Afterwards, the CL reagent, the emerging stream of luminol (presence of potassium ferrocyanide) and potassium ferricyanide flowed through the flow-cell and reacted with epinephrine adsorbed on the polymer to produce strong CL. The CL intensity responded linearly to the logarithm of the concentration of epinephrine within 5.0×10⁻⁹ to 1.0×10⁻⁷ mol/l range with a detection limit of 3×10⁻⁹ mol/l. The R.S.D. for 2.0×10⁻⁸ mol/l of epinephrine solution was less than 5% (n=7). Interference experiments demonstrated that the application of molecular imprinted polymer (MIP) to CL analysis could be improved the selectivity of the method greatly. Tests of the recovery for known amount of epinephrine in serum were carried out and satisfactory results were obtained. At the same time, the binding characteristic of the polymer to epinephrine was evaluated by the batch method and the dynamic method.     

A semi-automated method for fluorimetric determination of epinephrine and norepinephrine, without mutual interference, in single brain samples

A method is described for the separation and automated fluorimetric determination of epinephrine and norepinephrine in brain tissue extracts without mutual interference. The catecholamines are isolated and purified by extraction from activated alumina. Oxidation and rearrangement to their fluorescent lutines is carried out on two separate AutoAnalyzer manifolds and fluorescence is read in an Aminco-Bowman spectrophotofluorimeter. The interference by one amine with the determination of the other is less than 1% for determination of epinephrine in the presence of an equimolar concentration of norepinephrine, and, conversely, less than 4% for determination of norepinephrine. This eliminates the need for solution of simultaneous equations, the results of which are often misleading when the ratio of one amine to the other in brain exceeds 10:1. This method can be useful for rapid screening of psychoactive compounds affecting central and peripheral adrenergic stores.     

Determination of the solubility of inorganic salts by headspace gas chromatography

This work reports a novel method for determination of salt solubility using headspace gas chromatography. A very small amount of volatile compound (such as methanol) is added in the studied solution. Due to the molecular interaction in the solution, the vapor-liquid equilibrium (VLE) partitioning coefficient of the volatile species will change with different salt contents in the solution. Therefore, the concentration of volatile species in the vapor phase is proportional to the salt concentration in the liquid phase, which can be easily determined by headspace gas chromatography. Until the salt concentration in the solution is saturated, the concentration of volatile compound in the vapor phase will continue to increase further and a breakpoint will appear on the VLE curve. The solubility of the salts can be determined by the identification of the breakpoint. It was found that the measured solubility of sodium carbonate and sodium sulfate in aqueous solutions is slightly higher (about 6-7%) than those reported in the literature method. The present method can be easily applied to industrial solution systems.     

Simultaneous detection of epinephrine,uric acid,and ascorbic acid with graphene-modified electrode

A graphene-modified glassy carbon electrode was obtained via drop-casting method and applied to the simultaneous detection of epinephrine, uric acid, and ascorbic acid by cyclic voltammetry in a phosphate buffer solution (pH 3.0). The oxidation potentials of epinephrine, uric acid, and ascorbic acid were 0.484, 0.650, and 0.184 V at the graphene-modified glassy carbon electrode, respectively. The peak separation between epinephrine Pand uric acid, epinephrine and ascorbic acid, and uric acid and ascorbic acid was about 166, 300, and 466 mV, respectively. So, this graphene-modified electrode can be used for simultaneous determination of each component in a mixture.     

Cyclic Voltammetry Determination of Epinephrine with a Nano—gold Modified Glassy Carbon Electrode in the Presence of High Concentration Ascorbic Acid

As we know, there are usually some problems in electrochemical analysis with bare electrode to detect neurotransmitters. One is low electron transfer rate and the other is interfering compounds, such as ascorbic acid. So some kinds of modified electrodes have been successfully employed to promote the efficiency of electrochemical analysis1-4, such as Nafion modified electrode5,6 and SAM modified electrode7. They can attract positively-charged neurotransmitters while repulse negatively-cha…     

Fast enantiomeric separation of basis drugs by electrokinetic chromatography. Application to the quantitation of terbutaline in a pharmaceutical preparation

Electrokinetic chromatography (EKC) using micelles of bile salts alone or mixed with sodium dodecyl sulfate (SDS) and neutral, anionic, or cationic cyclodextrins (CDs) in the separation buffer has been employed in order to achieve fast enantiomeric separation of basic drugs. A study of the enantiomeric separation ability of these chiral selectors concerning four basic drugs (epinephrine, terbutaline, clenbuterol, and salbutamol) has been carried out under different experimental conditions. The best chiral selectors to perform the enantiomeric separation of these drugs were neutral beta-CD derivatives, specifically permethylated beta-CD PM-beta-CD. The effect of the PM-beta-CD concentration, temperature, and applied voltage on the enantiomeric resolution of the basic drugs was investigated. The use of a 25 mM ammonium acetate buffer (pH 5.0), 30 mM in PM-beta-CD together with an applied voltage of 20 kV and a temperature of 15 degrees C enabled the individual and fast enantiomeric separation of epinephrine, norepinephrine, terbutaline, clenbuterol, and salbutamol each one into its two enantiomers in less than 3 min. The EKC method was validated (precision and accuracy) to quantitate terbutaline in a pharmaceutical preparation, obtaining a limit of detection of 4 microg/mL.     

Simultaneous sensing of L-tyrosine and epinephrine using a glassy carbon electrode modified with nafion and CeO2 nanoparticles

An electrochemical sensor was developed and tested for detection of L-tyrosine in the presence of epinephrine by surface modification of a glassy carbon electrode (GCE) with Nafion and cerium dioxide nanoparticles. Fabrication parameters of a surfactant-assisted precipitation method were optimized to produce 2–3 nm CeO₂ nanoparticles with very high surface-to-volume ratio. The resulting nanocrystals were characterized structurally and morphologically by X-ray diffractometery (XRD), scanning and high resolution transmission electron microscopy (SEM and HR-TEM). The nanopowder is sonochemically dispersed in a Nafion solution which is then used to modify the surface of a GCE electrode. The electrochemical activity of L-tyrosine and epinephrine was investigated using both a Nafion-CeO₂ coated and a bare GCE. The modified electrode exhibits a significant electrochemical oxidation effect of L-tyrosine in a 0.2 M Britton-Robinson (B-R) buffer solution of pH 2. The electro-oxidation peak current increases linearly with the L-tyrosine concentration in the molar concentration range of 2 to 160 μM. By employing differential pulse voltammetry (DPV) for simultaneous measurements, we detected two reproducible peaks for L-tyrosine and epinephrine in the same solution with a peak separation of about 443 mV. The detection limit of the sensor (signal to noise ratio of 3) for L-tyrosine is ~90 nM and the sensitivity is 0.20 μA μM⁻¹, while for epinephrine these values are ~60 nM and 0.19 μA μM⁻¹. The sensor exhibited excellent selectivity, sensitivity, reproducibility and stability as well as a very good recovery time in real human blood serum samples.

Simultaneous electrochemical determination of L-tyrosine and epinephrine in blood plasma with Nafion-CeO₂/GCE modified electrode showing a 443 mV peak-to-peak potential difference between species oxidation peak currents.

     

Fabrication of a Nanostructure Based Electrochemical Sensor for Voltammetric Determination of Epinephrine,Uric Acid and Folic Acid

A carbon paste electrode (CPE) was modified by incorporation of graphene nano sheets and a ferrocene derivative. The modified electrode showed an excellent electrocatalytic effect on the oxidation of epinephrine. In phosphate buffer solution (PBS) of pH 7.0, the oxidation current increased linearly with concentration of epinephrine in the range of 0.05–550.0 µM and a detection limit (3σ) 27.0 nM was obtained for epinephrine. Then the modified electrode was used to determine epinephrine in an excess of uric acid and folic acid by SWV.     

Mass spectrometry monitoring of sevoflurane in the breathing circuit of an inhalation anesthesia machine

The results of controlling the amount of inhalation anesthetics sevoflurane in the patient breathing circuit of an inhalation anesthesia machine (IAM) by mass spectrometry are presented. A vacuum system with a differential chamber providing pressure difference in the range 1 × 10⁵–1.5 ×10⁻⁴ Pa was used to inject the studied gas sample from the delivery circuit into the mass spectrometer. The concentrations of the anesthetic obtained using mass and IR spectrometry are compared. The potency of mass spectrometry for monitoring the anesthetic gas in the real time mode is demonstrated. The time dependences of the concentration of the anesthetic gas corresponding to different periods of anesthesia are given.     

Dopamine Ion-Selective Electrode for Potentiometry in Pharmaceutical Preparations

 This paper reports the construction and evaluation of a dopamine sensitive electrode and its usefulness for the determination of this compound by direct potentiometry in pharmaceutical preparations. The electrode comprised a carboxylated poly(vinyl chloride) membrane based on β-cyclodextrine dissolved in 2-fluoro-2-nitrodiphenyl ether and using tetrakis(p-chlorophenyl) borate as a fixed anionic site. For comparison purposes membranes with similar composition but including normal high molecular weight PVC were also prepared. The electrodes including carboxylated poly(vinyl chloride) presented linear response within the concentration range of 5×10⁻⁵ and 10⁻¹ mol dm⁻³ of dopamine with a slope of about 59 mV decade⁻¹ of activity, in the pH range of 2 to 7.5 units. The response time was less than 15 seconds. Selectivity coefficients for different interferents including sodium, potassium, ammonium, lithium, epinephrine and norepinephrine were evaluated using the separated solution method and no significant interferences were observed. The electrode displays useful analytical characteristics for the direct determination of dopamine in pharmaceutical preparations. Results with an average recovery of 98.6±0.3% were obtained. Received May 28, 1998. Revision March 2, 1999.     

Enantiomeric Separation of Epinephrine and Salbutamol by Micellar Electrokinetic Chromatography Using β-Cyclodextrin as Chiral Additive

Introduction The development of chiral substances, especially inthe pharmaceutical field, places increasing demands onanalytical methods for the separation of these kinds ofisomers and the chiral purity control of drugs in phar-macokinetic studies. As the enantiomers of epinephrineand salbutamol have different pharmacological andtoxicological characteristics, separation and quantitationof the single enantiomers are required. Analytical methods used so far for the enantiomerseparation inclu…     

Formation of concentration gradient and its application to DNA capillary electrophoresis

A new method to introduce the concentration gradient into the capillary has been developed and its application to DNA capillary electrophoresis is presented. The concentration gradient produced by mixing 5% w/v polyacrylamide-co-poly(N-dimethylacrylamide) (PAM-co-PDMA) solution and 1 x Tris/N-tris(hydroxymethyl)methyl-3-amino-propanesulfonic acid/EDTA (TT) + 5 M urea buffer was successfully achieved by using two programmable syringe pumps with strict control of dead volume, flow rate, and pressure balance. This method has the advantages of high stability, reproducibility, and versatility. The column with concentration gradient greatly improved the resolution, especially for the large DNA fragments, due to a decrease in band width broadening with time. A column containing 2-4% w/v gradient in four steps had a longer read length, shorter separation time and better resolution (after 380 base) than that of 4% w/v single concentration polymer solution. The number of steps in the gradient had almost no effect on the performance. The change in the average concentration by relocating the position of the same step gradient, i.e., a combination of different low concentration to high concentration polymer solution ratios, resulted in a different migration time, read length and resolution.     

Joint effect of parameters of hydrochemical nepheline processing on alumina

A scheme of nepheline processing by a hydrochemical method was technically justified. In this process, silica is transformed into calcium hydrosilicate under autoclave conditions at a low concentration of caustic alkali. This allows the process at high heat regeneration and low steam flow for evaporation of a solution. A joint effect of temperature, leaching time, and concentration of caustic alkali solution was examined on an alumina recovery. Experiments were designed using a combined rotatable design of a second order.     

Simultaneous measurement of serotonin, catecholamines and their metabolites in cat and human plasma by in vitro microdialysis-microbore high-performance liquid chromatography with amperometric detection.

A method for the simultaneous measurement of norepinephrine, epinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, serotonin and 5-hydroxyindole-3-acetic acid in cat and human plasma by in vitro microdialysis-microbore high-performance liquid chromatography with electrochemical detection is described. The detection limit (signal-to-noise ratio = 3) is about 0.05-0.1 pg per injection. The volume of plasma samples required is very small (< 200 microliters), hence there is minimal blood loss in repeated blood sampling, especially in experiments using small animals. Within 15 min, a fast isocratic separation of these analytes by using a microbore reversed-phase ODS column is achieved, hence over 90 analyses can be performed in a single working day. As microdialysis per se is not destructive to plasma samples, the remaining plasma sample and perfusate can be repeatedly analysed for other substances. This simple, efficient and sensitive method can therefore be used as a routine clinical and basic research technique in the investigation of blood biogenic amines and their metabolites.     

An improved approach for the determination of plasma-free catecholamines by HPLC-ED: Application to normal and hypertensive patients

Summary A dual-step procedure for the rapid, quantitative isolation of free catecholamines (norepinephrine, epinephrine and dopamine) from plasma, using a little column of CM-Sephadex and alumina adsorption, is described. Sensitive high performance liquid chromatography is also discussed, employing an amperometric detector for the quantitative determination. The recovery of the three catecholamines, and of N-methyldopamine used as the internal standard, was about 70–80%; the detection limits were 2pg for norepinephrine, 3pg for epinephrine and 3pg for dopamine. The combination of the rather specific and easy to handle two-step sample clean-up procedure, the high resolving power of the chromatography and the high sensitivity of electrochemical detection provided a simple method for the determination of free catecholamines in plasma samples of normal and essential hypertensive subjects under different conditions (supine position for 45 min, standing for 5 and 10 min). It was found that a significant increase in epinephrine levels (P<0.01) occurred in hypertensive patients under the three conditions studied.     

Fully automated high-performance liquid chromatographic assay for the analysis of free catecholamines in urine

A totally automated and reliable high-performance liquid chromatographic method is described for the routine determination of free catecholamines (norepinephrine, epinephrine and dopamine) in urine. The catecholamines were isolated from urine samples using small alumina columns. A standard automated method for pH adjustment of urine before the extraction step has been developed. The extraction was performed on an ASPEC (Automatic Sample Preparation with Extraction Columns, Gilson). The eluate was collected in a separate tube and then automatically injected into the chromatographic column. The catecholamines were separated by reversed-phase ion-pair liquid chromatography and quantified by fluorescence detection. No manual intervention was required during the extraction and separation procedure. One sample may be run every 15 min, ca. 96 samples in 24 h. Analytical recoveries for all three catecholamines are 63-87%, and the detection limits are 0.01, 0.01, and 0.03 microM for norepinephrine, epinephrine and dopamine, respectively, which is highly satisfactory for urine. Day-to-day coefficients of variation were less than 10%.     

基于渗透脱水的自动化蛋白质结晶高通量筛选芯片

构建了一种基于渗透脱水模式的自动进样微流控结晶芯片. 该芯片通过真空预脱气将包含蛋白质和结晶剂的液滴自动分配至结晶微腔阵列中, 然后利用集成的一排包含不同浓度盐溶液的透析管道, 通过渗透脱水方式经一层聚二甲基硅氧烷(PDMS)膜实现液滴的逐渐浓缩, 使之趋于过饱和状态, 进而形成结晶. 此芯片可一次筛选较宽范围的过饱和状态, 实现蛋白质结晶的快速优化. 利用模式蛋白溶菌酶的结晶实验验证了该芯片的性能.