Photo-induced relaxation and proton transfer in some hydroxy naphthoic acids in polymers

Photophysical and photochemical properties of 3-hydroxy-2-naphthoic acid [3,2-HNA] and 1-hydroxy-2naphthoic acid [1,2-HNA] in different aprotic, protic, and ion exchange (Nafion) polymers have been described in this article. In both molecules, intramolecular hydrogen bond (IMHB) exists between OH and COOH functional groups. Both 3,2-HNA and 1,2-HNA form different emitting species in different polymeric media. Fluorescence characteristic of 3,2-HNA is found to depend on its concentration, nature of the microenvironment, and wavelength of excitation, while 1,2-HNA is less susceptible to these changes. 3,2-HNA exhibits dual fluorescence band (normal and large Stokes shifted) in aprotic and only a single large Stokes shifted fluorescence in protic polymers, while 1,2-HNA shows a single fluorescence band along with weak phosphorescence emission in these polymers. Both excited-state inter and intramolecular proton transfer (ESPT) take place in 3,2-HNA in aprotic and protic polymers, resulting in large Stokes shifted emission band. A competition between ESIPT and excimer formation is observed by the appearance of rise time on increasing the concentration of 3,2-HNA in protic polymer. In Nafion film, 3,2-HNA is present as a cationic as well as neutral species. The presence of extra protons in Nafion film facilitates excited-state intramolecular proton transfer (ESIPT) in the neutral species of 3,2-HNA and gives large Stokes shifted emission (10 500 cm(-1)). No such effect is observed in 1,2-HNA doped in Nafion film. It is observed that, depending on the position of the IMHB ring, the electronic spectra get modified and the strength of IMHB is affected by the micro-environment of the polymer which alters the photophysics of these molecules.     

Identification of hyaluronic acid oligosaccharides by direct coupling of capillary electrophoresis with electrospray ion trap mass spectrometry

A new method for the identification of oligosaccharides obtained by enzymatic digestion of hyaluronic acid (HA) with bacterial hyaluronidase (HA lyase, E.C. 4.2.2.1, from Streptococcus agalactiae) using online capillary electrophoresis/electrospray mass spectrometry (CE/ESI-MS) is presented. A fused-silica capillary coated with polyacrylamide was used with a 40 mM ammonium acetate buffer at pH 9.0 and a separation voltage of +30 kV applied to the inlet. Separation was achieved for oligosaccharides containing 4-16 monomers. The migration behavior follows the chain length of the oligomers, regardless of charge state. However, no linear relationship was found for the relation between mobility and chain length. Using an ion trap mass analyzer, complementary structural information was obtained by MS/MS and MS(n) experiments.     

Determination of heterocyclic aromatic amines by capillary high-performance liquid chromatography with diode array detection in ready-to-eat cooked ham treated with electron-beam irradiation

Heterocyclic aromatic amines (HAs) are a group of mutagenic and carcinogenic substances present in significant amounts in cooked meat and fish that can potentially be formed during food processing operations. This paper proposes a capillary liquid chromatography method with diode array detection for the trace-level determination of three HAs, namely, MeIQx (2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline), norharman (9H-pyrido[3,4-b]indole) and harman (1-methyl-9H-pyrido[3,4-b]indole), in ready-to-eat (RTE) cooked ham processed by electron-beam (accelerated electrons) irradiation to eliminate pathogenic microorganisms and to extend its shelf-life. The HAs selected have frequently been detected and quantified in a wide range of food and could be potential markers to indicate the presence of these toxic compounds. The method is based on the separation in an Inertsil C(8) capillary column (150 mm x 0.3-mm internal diameter, 3 microm) by gradient elution mode using a mixture of acetonitrile and 30 mM ammonium acetate pH 4.5 buffer as the mobile phase. Detection was at 250 and 265 nm and, to improve sensitivity, large injection volumes (20 microL) and on-column focusing techniques based on the injection of HA samples in low organic solvent strength solutions were employed. A simple and short solid-phase extraction and purification procedure was also optimized for sample preparation. Nonirradiated and irradiated RTE cooked ham samples at doses between 1 and 8 kGy were analyzed. HAs were not detected in any of the samples analyzed; so both types of samples were spiked at concentration levels in the range 5-25 ng g(-1), which may be found in meat products. The quality parameters of the method developed in the food matrix were established, and detection limits around 0.3 ng g(-1) were obtained. Spiked recoveries between 70 and 79% (n = 3 for each spiked level) relative standard deviations between 1 and 5% were also obtained, showing the effectiveness of the proposed method.     

On-column derivatization-capillary electrochromatography with o-phthalaldehyde/alkylthiol for assay of biogenic amines

The elution behaviors of the biogenic amines, histamine (HA) and its metabolite methyl histamine (MHA), were evaluated by means of on-column derivatization (OCD)-capillary electrochromatography (CEC) which employed a monolithic octadecylsilica (ODS) capillary column (20 cm of effective length x 50 microm of inner diameter). Five kinds of alkylthiols, e.g., 2- hydroxyethylthiol (or 2-mercaptoethanol (2-ME)), ethanethiol (ET), 1-propanethiol (1-PT), 2-methyl-1-propanethiol (2-MPT) and 1-butanethiol (1-BT) were separately presented at 5 mM each in the OCD-CEC separation run buffer consisting of 60% acetonitrile in 5 mM o-phthalaldehyde (OPA)-10 mM borate buffer (pH 10). When 2-ME was present in the run buffer solution, both derivatives corresponding to HA and MHA migrated separately, but closely together through the capillary column. Replacement of 2-ME with 1-BT in the run buffer solution caused a delay in their elution profiles on the electrochromatogram and the separation between those two peaks became remarkably improved. The elution times of HA and MHA followed the increase in alkyl chain length or hydrophobicity of thiol, 1-BT > 2-MPT > 1-PT > ET > 2-ME. Performance of on-line preconcentrations of HA and MHA was also evaluated by varying the electrokinetic injection voltage from 1 kV to 8 kV. The peak area counts corresponding to HA recorded about 50 times higher when 2 kV was applied for 240 s to a 0.1 mM HA solution than when 8 kV was applied for 5 s. This method was next applied to a sample of human urine spiked with HA and MHA at levels of 0.1 microM each. Although HA and MHA peaks were not identifiable among the peaks corresponding to the materials in the urine matrix when OPA/2-ME was employed in a run buffer for the OCD-CEC, the separation and identification of their peaks became possible by replacing 2-ME with 1-BT in the run buffer solution.     

An experimental and theoretical investigation of the photophysics of 1-hydroxy-2-naphthoic acid

Photophysical and photochemical properties of 1-hydroxy-2-naphthoic acid (1,2-HNA) have been investigated experimentally by steady state and time domain fluorescence measurements and theoretically by Hartree-Fock (HF), configuration interaction at the single excitation (CIS) level, density functional theoretic (DFT), and semiempirical (AM1) methods. 1,2-HNA exhibits normal fluorescence that depends on its concentration, nature of the solvent, pH, temperature, and wavelength of excitation. It seems to form different emitting species in different media, akin to 3-hydroxy-2-naphthoic acid (3,2-HNA). The large Stokes shifted emission observed at pH 13 is attributed to species undergoing excited-state intramolecular proton transfer. Nonradiative transition seems to increase on protonation and decrease on deprotonation. AM1(PECI=8) calculations predict the absorption maximum (lambda(max) = 335.9 nm) in reasonable agreement with experiment (lambda(max) = 352 nm) for the neutral 1,2-HNA. They also predict a red shift in absorption on protonation and a blue shift on deprotonation as observed experimentally. CIS calculations tend to overestimate the energy gap and hence underestimate the absorption maxima between the ground and the excited electronic states of 1,2-HNA and its protonated and deprotonated forms. However, they do predict correctly that the excited state intramolecular proton transfer is likely to occur in the deprotonated form of 1,2-HNA and not in the neutral and the protonated forms. A single minimum is found in the potential energy profile for the ground state as well as the first excited state of 1,2-HNA and its protonated species. In contrast, a double minimum with a nominal barrier in between is predicted for the ground state and also the first three excited states of the deprotonated species. The keto form of the deprotonated species is found to be slightly less stable than the enol form in all the states investigated.     

Analysis of illicit drugs by nonaqueous capillary electrophoresis and electrochemical detection

Nonaqueous capillary electrophoresis (NACE) was applied to the determination of illicit drugs. The complete separation of amphetamine, methamphetamine, 3,4-methylene dioxy amphetamine (MDA), 3,4-methylene dioxy methamphetamine (MDMA), mescaline, cocaine and benzoylecgonine was obtained using an acetonitrile based buffer solution containing 10 mM sodium acetate and 1 M acetic acid. Electrochemical detection using a Pt microdisk electrode set to a potential of +1.8 V was found to be selective for MDA, MDMA and mescaline. The detection limits for these compounds were in the low ng/mL range which is between 2 and 3 orders of magnitude lower compared to UV-detection.     

Analysis of illicit drugs by nonaqueous capillary electrophoresis and electrochemical detection

Nonaqueous capillary electrophoresis (NACE) was applied to the determination of illicit drugs. The complete separation of amphetamine, methamphetamine, 3,4-methylene dioxy amphetamine (MDA), 3,4-methylene dioxy methamphetamine (MDMA), mescaline, cocaine and benzoylecgonine was obtained using an acetonitrile based buffer solution containing 10 mM sodium acetate and 1 M acetic acid. Electrochemical detection using a Pt microdisk electrode set to a potential of +1.8 V was found to be selective for MDA, MDMA and mescaline. The detection limits for these compounds were in the low ng/mL range which is between 2 and 3 orders of magnitude lower compared to UV-detection.     

分子印迹Mn掺杂ZnS量子点磷光法检测烟酸转化液中的6-羟基烟酸

以6-羟基烟酸(6-HNA)为模板,采用分子印迹溶胶-凝胶法合成了具有良好光学性质的印迹Mn掺杂Zn S量子点磷光传感器(MIP-QDs)。红外光谱(FT-IR)和扫描电镜(SEM)测试表明,印迹聚合物成功接枝在纳米传感器表面。吸附试验显示,MIP-QDs对6-HNA的亲和性与选择性显著高于其结构类似物阔马酸和烟酸,具备作为检测探针的潜力。在最优条件下,基于6-HNA可选择性猝灭MIP-QDs磷光而建立了灵敏、简便的检测微量6-HNA的光学新方法。当6-HNA浓度在3.4~67.0μmol/L范围内,MIP-QDs的磷光猝灭率P_0/P随浓度变化的关系符合Stern-Volmer方程(r~2=0.996 5),检出限为1.03μmol/L。实际样品测定显示,6-HNA的回收率为91.0%~103.9%,相对标准偏差不超过4.5%,检测结果与HPLC法相一致(P0.05),方法具有良好的准确性和可行性。     

Separation and identification of photodegradation products of benzoic acid by capillary zone electrophoresis

A capillary zone electrophoresis (CZE) method was developed for separation and identification of photodegradation products of benzoic acid under irradiation at a wavelength of 300 nm. Parameters such as run buffer, applied voltage and injection time were optimized for the separation of benzoic acid and its photodegradation products. Linearity, limit of detection, and repeatability of migration time as well as peak area of the method were examined. Four reaction products, including salicylic acid, 3-hydroxybenzoic acid, 4-hydroxybenzoic acid and 3,4-dihydroxybenzoic acid have been separated and identified by spiking the known compounds into the irradiated samples using the CZE method developed. The confirmation of the reaction products is one of the key steps for proposing the possible reaction mechanisms involved in the photodegradation of benzoic acid.     

Non-aqueous capillary electrophoresis with diode array and electrospray mass spectrometric detection for the analysis of selected steroidal alkaloids in plant extracts

A capillary electrophoresis (CE) assay has been developed for the quantitation and determination of the impurity profile of the potassium channel blockers 3,4-diaminopyridine and 4-aminopyridine. The compounds were separated from related substances using a capillary of 30 cm effective length, a 50 mM phosphate buffer, pH 2.5 and an applied voltage of 25 kV. The assay was validated with respect to specificity, linearity, range, limits of quantitation and detection, precision and robustness. The method allows the detection and quantitation of impurities at the 0.05% level. The feasibility of the assay was demonstrated by analyzing a commercial sample of 3,4-diaminopyridine. All known related substances could be detected in this sample with the present CE method.     

Post-column derivatization capillary electrochromatography for detection of biogenic amines in tuna-meat

A system to perform post-column derivatization capillary electrochromatography (CEC) was developed for the first time. The system mainly included a 4-microm (O.D.) silica packed column (200 mm effective length x 0.1 mm inner diameter I.D.) with micro-magnetic particles (MMPs) frits, a T-junction connector, an in-line fluorescence detector and a high-voltage power supply. The system was evaluated by using histamine (HA) as a standard biogenic amine for this study. A 5 microM HA solution was loaded at the anodic site of the capillary column by applying 3 kV for 5s. Then, HA was electrophoretically eluted with a 20mM phosphate buffer (pH 7) by applying 3 kV, and was derivatized with 3mM o-phthalaldehyde (OPA)/N-acetylcysteine (NAC) in 100 mM borate (pH 10), which was continuously delivered through the reagent-loading capillary tube by gravity into the T-junction connector. HA derivative was finally detected with the in-line fluorescence detector (lambda(Ex)=340 nm, lambda(Em)=450 nm) at 9.7 min after sample loading. To test the utility of this system, it was next employed for its ability to detect the presence of HA and other kinds of biogenic amines, including cadaverine (Cad), spermidine (Spm) and tyramine (Tyr) in tuna-meat, once the validity of the method had been confirmed.     

Novel Dispersion of MWCNTs in Polystyrene Polymer Induced by the Addition of 3-Hydroxy-2-Napthoic Acid

This article reports an extensive investigation of the unique dispersion behavior of solutions with multi-walled carbon nanotubes (MWCNTs) and 3-hydroxy-2-napthoic acid (β-HNA) in tetrahydrofuran (THF) solvent, which results into a multifold enhancement in the electrical properties of polystyrene (PS). A number of solutions with 0.4% of MWCNTs (w/v) and β-HNA (0–1%, w/v) in THF were prepared separately. MWCNTs precipitated out in THF solvent shortly after the preparation and formed two distinct phase regions (2φ). Gratifyingly, addition of β-HNA solution to the MWCNTs solution offered an unprecedented enhancement in the dispersion of MWCNTs. Such dispersion in solutions with only 0.02% β-HNA (w/v) was found to be stable up to 2 weeks at room temperature. FTIR spectroscopy was incorporated to illustrate the adsorption of β-HNA onto the surface of carbon nanotubes (CNTs). After this successful dispersion, nanocomposites solutions comprising of 0.067% β-HNA (w/v), 6.7% PS (w/v), and varying concentrations of MWCNTs (0–0.33%, w/v) were prepared. A remarkable dispersion behavior of MWCNTs in the presence of polymer was also observed. Finally, thin films made up of consistent polystyrene/β-HNA concentrations and increasing amounts of MWCNTs were prepared by casting technique to investigate the influence of dispersion on the electrical properties of the film. The dispersion significantly affected the DC electrical conductivity and incorporation of 5% MWCNTs elevated the electrical conductivity up to 10 orders of magnitude with respect to neat PS.     

Poly(methacrylic acid-ethylene glycol dimethacrylate) monolith in-tube solid-phase microextraction applied to simultaneous analysis of some amphetamine derivatives in urine by capillary zone electrophoresis

A method based on in-tube solid-phase microextraction and capillary zone electrophoresis (CZE) was proposed for simultaneously determining four amphetamines (amphetamine, methamphetamine, 3,4-methylenedioxyamphetamine, and 3,4-methylenedioxymethamphetamine) in urine. A poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column, which can provide sufficient extraction efficiency, was introduced for the extraction of amphetamines from urine samples. The hydrophobic main chains and acidic pendant groups of the monolithic column make it a superior material for extraction of basic analytes from aqueous matrix. After extraction, the samples were analyzed by CZE. The best separation was achieved using a buffer composed of 0.1 M disodium hydrogen phosphate (adjusted to pH 4.5 with 1 M hydrochloric acid) and 20% methanol v/v, with a temperature and voltage of 25 degrees C and 20 kV, respectively. By applying electrokinetic injection with field-amplified sample stacking, detection limits of 25-34 microg/L were achieved. Excellent method of reproducibility was found over a linear range of 0.1-5 mg/L. Determination of these analytes from abusers' urine sample was also demonstrated.     

Determination of urinary catecholamines with capillary electrophoresis after solid-phase extraction

The stabilities of 3,4-dihydroxybenzylamine (DHBA), dopamine, 3-methoxytyramine, normetanephrine and metanephrine standards under acid, base and enzymatic hydrolysis conditions were studied. Basic incubation media were not suitable for 3,4-dihydroxy compounds, but acid and enzymatic hydrolysis conditions were applicable to all the compounds. The results of acid and enzymatic hydrolysis were comparable and the enzymatic hydrolysis was applied to a urine matrix. A method including solid-phase extraction (SPE) with a copolymer sorbent was developed for purification of the urine samples. Due to poor recovery of DHBA, the most frequently used internal standard in catecholamine analysis, this compound was replaced with the 3-O-methoxy structure. The recoveries of the compounds in spiked urine samples in SPE were between 96.4 and 124.4%. The repeatability of the combination of enzymatic hydrolysis and SPE pretreatment was good for all the compounds, except for dopamine and 3-methoxytyramine due to some matrix compounds still interfering with the separation. The analyses were performed with capillary electrophoresis in an ammonium acetate buffer with UV detection. The validation data for the compounds including limit of detection, limit of quantification, linearity and repeatability of the method are presented.     

Study on the Effect of Moderate Exercise on Lactic Acid Content in Breast Milk by Indirect CE with Amperometric Detection

An indirect high-performance capillary electrophoresis with amperometric detection (CE-AD) method has been developed for determination of lactic acid (LA) in body fluids of lactating postpartum women. Several important factors such as the running buffer additive and concentration, the working electrode potential, the pH value and concentration of the running buffer, the separation voltage and injection time were investigated. Under the optimum conditions, LA could be well separated with co-existing interferences including uric acid (UA) in real samples in a 90-cm-length capillary at separation voltage of 12 kV in 4.0 × 10^?6 g mL^?1 3,4-dihydroxybenzylamine (DHBA)/40 mmol L^?1 H₃BO₃?CNa₂B₄O₇ buffer (pH 7.8). The linearity between peak current and concentration of LA was over three orders of magnitude with detection limit of 5.00 × 10^?7 g mL^?1 (S/N = 3). This proposed method has been successfully used to study the effects of moderate exercise on LA content in breast milk and urine samples of lactating postpartum women, and assay results showed that LA content in breast milk can return to normal level through 60 min rest without decreasing acceptance by breast-feeding infants, although the LA level did increase by 4?C6 times in both breast milk and urine samples at 10 min after moderate exercise.     

Evaluation of the stability constants of acidic and basic organic substances with 18-crown-6 and β-cyclodextrin using capillary zone electrophoresis

The possibility of evaluating stability constants in the analyte-macrocycle system by capillary zone electrophoresis was exemplified in the separation of a mixture of acids (homovanillic acid, vanilmandelic acid, 5-hydroxyindole-3-acetic acid, and 3,4-dihydroxyphenylalanine) or bases (adrenalin, noradrenalin, dopamine, serotonin, and metanephrine) with the addition of β-cyclodextrin or 18-crown-6, respectively, to the buffer electrolyte. The information content of the constants obtained in the separation of biologically important mixtures for the determination of optimum concentrations of the complex-forming agents in the running buffer is discussed.     

Analysis of atrazine, terbutylazine and their N-dealkylated chloro and hydroxy metabolites by solid-phase extraction and gas chromatography-mass spectrometry and capillary electrophoresis-ultraviolet detection

Solid-phase extraction (SPE) with the styrene-divinylbenzene adsorbent LiChrolut EN was investigated for the extraction of the s-triazine herbicides atrazine and terbutylazine, their polar N-dealkylated degradation products deethylatrazine (DEA), deisopropylatrazine (DIA) and deethylterbutylazine (DET) and for the hydrophilic hydroxytriazine degradation products (HTDPs) hydroxyatrazine (HA), hydroxyterbutylazine (HT), deethylhydroxyatrazine (DEHA), deisopropylhydroxyatrazine (DIHA) and deethyldeisopropylhydroxyatrazine (ameline). The optimum pH value for the extraction of the HTDPs from fortified tap water at 2 micrograms/l is 3.0. Recovery values with 200 mg LiChrolut EN are > 80% for HA, HT, DEHA and 30% for DIHA from 200 ml spiked tap and river water. Atrazine, terbutylazine, DEA, DIA and DET are quantitatively extracted by LiChrolut EN. The chlorotriazines are analyzed by GC-MS and the HTDPs by capillary zone electrophoresis (CZE) and micellar electrokinetic capillary chromatography (MECC) with an acetate buffer at pH 4.6 or a sodium borate-sodium dodecyl sulfate buffer at pH 9.3. The combined method of SPE enrichment and CE analysis allows the determination of HTDPs in the low microgram/l range.     

Investigating the binding interaction of azur A with hyaluronic acid via spectrophotometry and its analytical application

The interaction between azur A (AA) and hyaluronic acid (HA) at AA concentrations from 3.430×10^–5 to 8.575×10^–5 M and sodium chloride concentrations from 0 to 0.01 M was investigated spectrophotometrically at 620 nm at temperatures from 0 to 50 °C. AA was shown to be a useful spectroscopic probe for detecting carboxyl groups in HA macromolecules. The interaction between AA and HA was temperature sensitive and little AA–HA interaction was observed at temperatures higher than 30 °C. The interaction of HA with AA was seen to be electrostatic in nature. The maximum binding number decreased with decreasing NaCl concentration, and the absorbance sensitivity decreased with increasing NaCl concentration in aqueous solution. Self-interference from the AA in the AA–HA interaction caused an overestimate of the molar mass of hyaluronic acid. An improved method was proposed to estimate the molar mass of HA, and a molar mass of 1.219×10⁶ Da was obtained with this improved method for HA.     

Simultaneous determination of electroactive and non-electroactive food preservatives by novel capillary electrophoresis with amperometric detection

A novel capillary electrophoresis and amperometric detection method was achieved by adding an electroactive additive (3,4-dihydroxybenzylamine, 3,4-DHBA) to the running buffer, so that both electroactive and non-electroactive food preservatives were simultaneously determined. Under the selected optimum conditions, four electroactive preservatives (methylparaben, ethylparaben, propylparaben and butylparaben) and two non-electroactive preservatives (potassium sorbate and sodium lactate) were well separated and sensitively detected with detection limits (S/N = 3) ranging from 1.06 × 10⁻⁸ to 2.73 × 10⁻⁶ g mL⁻¹. This method has been successfully employed for the determination of both electroactive and non-electroactive preservatives in several food commodities.     

Rapid separation and laser-induced fluorescence detection of mutated DNA by capillary electrophoresis in a self-coating, low-viscosity polymer matrix

The detection of point and other simple mutations in DNA is important for cancer research and diagnosis and other biological studies. Capillary electrophoresis has been successfully used for separating DNA fragments. However, a low-viscosity polymer sieving buffer for DNA separation with on-line coating has never been reported. In this paper, a new method using capillary electrophoresis with on-line coating and laser-induced fluorescence detection (CE-LIF) for screening for point or simple DNA mutations has been demonstrated. The method uses an on-line dynamic coating technique that increases capillary lifetime and analysis reproducibility, and employs a low-viscosity polymer solution, which allows the user to rinse the capillary rapidly and refill with polymer solution easily. Experiments proved that the additives in the separation buffer for on-line capillary coating do not affect the separation efficiency of the running buffer, and do not interfere with the formation of hydrogen-bonded network between boric acid, mannitol and hydroxypropylmethylcellulose polymers. The stability of the dynamically coated capillary was quantitatively studied; the capillary lifetime was increased 6- to 7-fold compared with that of permanently coated CE columns. Standard DNA fragments containing mutations, with sizes of 209, 219, and 338 bps, were successfully separated and detected with this system, after the mutated DNA fragments were cleaved by CEL-I endonuclease. The technique is very sensitive for the size-separation of low-range, middle-range, and high-range DNA fragments. Results were compared with the HPLC methods developed by Transgenomic, Inc. and were in good agreement. The method should be applicable to mutation detection for all relevant biological and clinical studies. The factors influencing separations and the stability of dynamic capillary coatings are also discussed in the paper.