Determination of enzyme activity inhibition by FTIR spectroscopy on the example of fructose bisphosphatase

A mid-infrared enzymatic assay for label-free monitoring of the enzymatic reaction of fructose-1,6-bisphosphatase with fructose 1,6-bisphosphate has been proposed. The whole procedure was done in an automated way operating in the stopped flow mode by incorporating a temperature-controlled flow cell in a sequential injection manifold. Fourier transform infrared difference spectra were evaluated for kinetic parameters, like the Michaelis–Menten constant (K _M) of the enzyme and V _max of the reaction. The obtained K _M of the reaction was 14 ± 3 g L⁻¹ (41 μM). Furthermore, inhibition by adenosine 5′-monophosphate (AMP) was evaluated, and the K _M^App value was determined to be 12 ± 2 g L⁻¹ (35 μM) for 7.5 and 15 μM AMP, respectively, with V _max decreasing from 0.1 ± 0.03 to 0.05 ± 0.01 g L⁻¹ min⁻¹. Therefore, AMP exerted a non-competitive inhibition.     

Thermomechanical properties of cured isophtalic polyester resin modified with poly(ε-caprolactone)

The effect of a low profile additive, poly(ε-caprolactone) (PCL), on the thermal and mechanical properties of unsaturated polyester resins (UP) was investigated by differential scanning calorimetry (DSC), dynamic mechanical thermal analysis (DMTA), and tensile tests. The morphology of the systems has been studied by scanning electronic microscopy (SEM). Two PCL molecular mass were selected (PCL2: M _n = 2000 g mol⁻¹ and PCL50: M _n = 50000 g mol⁻¹) to analyze the influence of the molecular mass and the content of PCL on the UP resins and to establish the relation between thermomechanical behavior and morphology. DSC and DMTA glass transition temperatures (T _g) of the UP cured samples containing PCL indicate that PCL2 is miscible with UP whereas for UP + PCL50 system, T _g values are very close to the ones corresponding to neat UP. Besides in UP + PCL2 systems, one phase morphology is observed in which PCL2 would act as solvent of the reacting mixture along curing process; however, UP + PCL50 systems present phase-separated morphology. The presence of PCL2 and PCL50 in UP resin leads to a decrease of the tensile strength and the Young′s modulus as much notorious as the PCL concentration increases. For UP + PCL2 system the elongation at fracture increases in relation to neat UP, increasing as well with the PCL content.     

Scale up and Application of Biosurfactant from <Emphasis Type="Italic">Bacillus subtilis</Emphasis> in Enhanced Oil Recovery

There is a lack of fundamental knowledge about the scale up of biosurfactant production. In order to develop suitable technology of commercialization, carrying out tests in shake flasks and bioreactors was essential. A reactor with integrated foam collector was designed for biosurfactant production using Bacillus subtilis isolated from agricultural soil. The yield of biosurfactant on biomass (Y _p/x), biosurfactant on sucrose (Y _p/s), and the volumetric production rate (Y) for shake flask were obtained about 0.45 g g⁻¹, 0.18 g g⁻¹, and 0.03 g l⁻¹ h⁻¹, respectively. The best condition for bioreactor was 300 rpm and 1.5 vvm, giving Y _x/s, Y _p/x, Y _p/s, and Y of 0.42 g g⁻¹, 0.595 g g⁻¹, 0.25 g g⁻¹, and 0.057 g l⁻¹ h⁻¹, respectively. The biosurfactant maximum production, 2.5 g l⁻¹, was reached in 44 h of growth, which was 28% better than the shake flask. The obtained volumetric oxygen transfer coefficient (K _L a) values at optimum conditions in the shake flask and the bioreactor were found to be around 0.01 and 0.0117 s⁻¹, respectively. Comparison of K _L a values at optimum conditions shows that biosurfactant production scaling up from shake flask to bioreactor can be done with K _L a as scale up criterion very accurately. Nearly 8% of original oil in place was recovered using this biosurfactant after water flooding in the sand pack.     

Electro-catalysis by immobilised human flavin-containing monooxygenase isoform 3 (hFMO3)

Human flavin-containing monooxygenases are the second most important class of drug-metabolizing enzymes after cytochromes P450. Here we report a simple but functional and stable enzyme-electrode system based on a glassy carbon (GC) electrode with human flavin-containing monooxygenase isoform 3 (hFMO3) entrapped in a gel cross-linked with bovine serum albumin (BSA) by glutaraldehyde. The enzymatic electrochemical responsiveness is characterised by using well-known substrates: trimethylamine (TMA), ammonia (NH₃), triethylamine (TEA), and benzydamine (BZD). The apparent Michaelis–Menten constant (K′_M) and apparent maximum current (I′_max) are calculated by fitting the current signal to the Michaelis–Menten equation for each substrate. The enzyme-electrode has good characteristics: the calculated sensitivity was 40.9 ± 0.5 mA mol⁻¹ L cm⁻² for TMA, 43.3 ± 0.1 mA mol⁻¹ L cm⁻² for NH₃, 45.2 ± 2.2 mA mol⁻¹ L cm⁻² for TEA, and 39.3 ± 0.6 mA mol⁻¹ L cm⁻² for BZD. The stability was constant for 3 days and the inter-electrode reproducibility was 12.5%. This is a novel electrochemical tool that can be used to investigate new potential drugs against the catalytic activity of hFMO3.     

Catalytic and Thermodynamic Characterization of Endoglucanase (CMCase) from <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> cmc-1

Monomeric extracellular endoglucanase (25 kDa) of transgenic koji (Aspergillus oryzae cmc-1) produced under submerged growth condition (7.5 U mg⁻¹ protein) was purified to homogeneity level by ammonium sulfate precipitation and various column chromatography on fast protein liquid chromatography system. Activation energy for carboxymethylcellulose (CMC) hydrolysis was 3.32 kJ mol⁻¹ at optimum temperature (55 °C), and its temperature quotient (Q ₁₀) was 1.0. The enzyme was stable over a pH range of 4.1–5.3 and gave maximum activity at pH 4.4. V _max for CMC hydrolysis was 854 U mg⁻¹ protein and K _m was 20 mg CMC ml⁻¹. The turnover (k _cat) was 356 s⁻¹. The pK _a1 and pK _a2 of ionisable groups of active site controlling V _max were 3.9 and 6.25, respectively. Thermodynamic parameters for CMC hydrolysis were as follows: ΔH* = 0.59 kJ mol⁻¹, ΔG* = 64.57 kJ mol⁻¹ and ΔS* = −195.05 J mol⁻¹ K⁻¹, respectively. Activation energy for irreversible inactivation ‘E _a(d)’ of the endoglucanase was 378 kJ mol⁻¹, whereas enthalpy (ΔH*), Gibbs free energy (ΔG*) and entropy (ΔS*) of activation at 44 °C were 375.36 kJ mol⁻¹, 111.36 kJ mol⁻¹ and 833.06 J mol⁻¹ K⁻¹, respectively.     

Production of Lipids Containing High Levels of Docosahexaenoic Acid by a Newly Isolated Microalga, <Emphasis Type="Italic">Aurantiochytrium</Emphasis> sp. KRS101

In the present study, a novel oleaginous Thraustochytrid containing a high content of docosahexaenoic acid (DHA) was isolated from a mangrove ecosystem in Malaysia. The strain identified as an Aurantiochytrium sp. by 18S rRNA sequencing and named KRS101 used various carbon and nitrogen sources, indicating metabolic versatility. Optimal culture conditions, thus maximizing cell growth, and high levels of lipid and DHA production, were attained using glucose (60 g l⁻¹) as carbon source, corn steep solid (10 g l⁻¹) as nitrogen source, and sea salt (15 g l⁻¹). The highest biomass, lipid, and DHA production of KRS101 upon fed-batch fermentation were 50.2 g l⁻¹ (16.7 g l⁻¹ day⁻¹), 21.8 g l⁻¹ (44% DCW), and 8.8 g l⁻¹ (40% TFA), respectively. Similar values were obtained when a cheap substrate like molasses, rather than glucose, was used as the carbon source (DCW of 52.44 g l⁻¹, lipid and DHA levels of 20.2 and 8.83 g l⁻¹, respectively), indicating that production of microbial oils containing high levels of DHA can be produced economically when the novel strain is used.     

Effect of organic load on the performance and methane production of an AnSBBR treating effluent from biodiesel production

Currently, there is an increasing demand for the production of biodiesel and, consequently, there will be an increasing need to treat wastewaters resulting from the production process of this biofuel. The main objective of this work was, therefore, to investigate the effect of applied volumetric organic load (AVOL) on the efficiency, stability, and methane production of an anaerobic sequencing batch biofilm reactor applied to the treatment of effluent from biodiesel production. As inert support, polyurethane foam cubes were used in the reactor and mixing was accomplished by recirculating the liquid phase. Increase in AVOL resulted in a drop in organic matter removal efficiency and increase in total volatile acids in the effluent. AVOLs of 1.5, 3.0, 4.5 and 6.0 g COD L⁻¹ day⁻¹ resulted in removal efficiencies of 92%, 81%, 67%, and 50%, for effluent filtered samples, and 91%, 80%, 63%, and 47%, for non-filtered samples, respectively, whereas total volatile acids concentrations in the effluent amounted to 42, 145, 386 and 729 mg HAc L⁻¹, respectively. Moreover, on increasing AVOL from 1.5 to 4.5 g COD L⁻¹ day⁻¹ methane production increased from 29.5 to 55.5 N mL CH₄ g COD⁻¹. However, this production dropped to 36.0 N mL CH₄ g COD⁻¹ when AVOL was increased to 6.0 g COD L⁻¹ day⁻¹, likely due to the higher concentration of volatile acids in the reactor. Despite the higher concentration of volatile acids at the highest AVOL, alkalinity supplementation to the influent, in the form of sodium bicarbonate, at a ratio of 0.5–1.3 g NaHCO₃ g COD_fed⁻¹, was sufficient to maintain the pH near neutral and guarantee process stability during reactor operation.     

Precipitation polymerization of <Emphasis Type="Italic">N-tert</Emphasis>-butylacrylamide in water producing monodisperse polymer particles

The precipitation polymerizations of N-tert-butylacrylamide (NtBAM) in water are demonstrated; for example, the polymerization with potassium peroxodisulfate using a 15 g L⁻¹ (118 mmol L⁻¹) concentration of NtBAM in the feed ([NtBAM]₀) was performed at 70 °C for 12 h, quantitatively producing poly(N-tert-butylacrylamide) particles with a number-average diameter (d _n) of 203 nm and a coefficient of variation (C _v) of 4.7%. The particle sizes were controlled in the d _ns range between 75 and 494 nm by changing the monomer feeds or adding an electrolyte such as NaCl. The solid contents in the resulting aqueous latex solutions ranged from 0.1 to 1.5%, whereas it increased to 4.8% by applying a “shot-growth” technique. The polymerization in water under a somewhat unique condition is described, which was started from a heterogeneous system due to the presence of significantly large amounts of monomers ([NtBAM]₀ = 50 g L⁻¹). This also provided monodisperse latexes with the d _n of 370 nm in 96% yield, in which the solid content reached 4.9%.     

Flavonol Derivatives for Determination of Cr(III) and W(VI)

 Simple, rapid, sensitive and selective methods for the determination of Cr(III) and W(VI) with flavonol derivatives in the presence of surface-active agents are proposed. In the pH ranges 3.4–4.2 and 1.9–2.5, the molar absorptivities of Cr(III)-morin-emulsifier S (EFA) and W(VI)-morin-polyvinylpyrrolidone (PVP) systems are 1.13×10⁵ and 2.13×10⁴ L mol⁻¹ cm⁻¹ at 435 and 415 nm, respectively. The Cr(III)-quercetin-PVP and W(VI)-quercetin-cetylpyridinium bromide (CPB) systems are formed in the pH ranges 4–4.6 and 2.2–2.8 with molar absorptivities 1.02×10⁵ and 9.02×10⁴ L. mol⁻¹ cm⁻¹ at 441 and 419 nm, respectively. The linear dynamic ranges for the determination of Cr(III) and W(VI) with morin in the presence of EFA and PVP are 0.03–0.46 and 0.71–8.1 μg mL⁻¹, respectively. The corresponding ranges with quercetin are 0.04–0.54 and 0.14–2.1 μg mL⁻¹ of Cr(III) and W(VI), respectively. The r.s.d (n = 10) for the determination of 0.25 and 3.7 μg mL⁻¹ of Cr(III) and W(VI) with morin and their detection limits are 0.88 and 0.99% and 0.016 and 0.63 μg mL⁻¹, respectively. Using quercetin, the r.s.d (n = 10) for 0.22 and 1.2 μg mL⁻¹ of Cr(III) and W(VI) and their detection limits are 0.92 and 0.91% and 0.015 and 0.08 μg mL⁻¹, respectively. The critical evaluation of the proposed methods is performed by statistical analysis of the experimental data. The proposed methods are applied to determine Cr in steel, non-ferrous alloys, wastewater and mud filtrate and to the determination of W in steel. Received March 8, 1999. Revision January 21, 2000.     

Effects of Temperature and Common Ions on Binding of Puerarin to BSA

The effects of temperature and common ions on binding of puerarin to bovine serum albumin (BSA) are investigated. The binding constants (K _a) between puerarin and BSA are 1.13×10⁴ L⋅mol⁻¹ (20 °C) and 1.54×10⁴ L⋅mol⁻¹ (30 °C), and the number of binding sites (n) is (0.95±0.02). However, at a higher temperature (40 °C) the stability of the puerarin–BSA system decreases, which results in a lower binding constant (1.58×10³ L⋅mol⁻¹) and number of binding sites (n=0.73) of the puerarin–BSA system. However, the presence of Cu²⁺ and Fe³⁺ ions increases the binding constants and the number of binding sites in the puerarin–BSA complex.     

Production and analytical characterization of neopterin immunoreagents for biosensor developments

Neopterin is a valuable biomarker of cellular immunity associated with various pathological situations such as viral and bacterial infections, autoimmune, cardiovascular, neurodegenerative and malignant disorders. To produce specific antibodies against neopterin for a rapid multi-biomarker-based diagnosis, a novel hapten derivative was synthesized and attached to carrier proteins. The thoroughly characterized conjugates were used for immunization of BALB/c mice and rabbits. The produced monoclonal antibody reached in both direct and indirect enzyme-linked immunosorbent assay (ELISA) format LoD of 0.18 and 0.45 μg L⁻¹, respectively, and was a superior immunoreagent for further biosensor developments with regard to assay sensitivity and material availability. The best polyclonal antibody was somewhat more sensitive in direct ELISA with LoD of 0.05 μg L⁻¹. The optimized ELISA method was evaluated with blood samples collected from patients with renal insufficiency, patients with sepsis, patients without confirmed clinical diagnosis, and healthy volunteers. In plasma samples, neopterin concentrations ranging from 3.2 to 103 μg L⁻¹ could be determined with the monoclonal ELISA whereas twofold lower results were obtained with the polyclonal ELISA. A satisfactory correlation of results was found between the polyclonal ELISA and IBL Neopterin ELISA kit within the concentration range 0.5–16 μg L⁻¹ (R = 0.874; n = 40), and slightly lower correlation was found for monoclonal-based ELISA (R = 0.819; n = 40). These data show that the generated antibodies may be used as functional analytical reagents for the integration into multianalyte biochip detection systems.     

Development of a liquid chromatography-mass spectrometry method for the determination of ursolic acid in rat plasma and tissue: application to the pharmacokinetic and tissue distribution study

A fast and sensitive liquid chromatography–mass spectrometry method was developed for the determination of ursolic acid (UA) in rat plasma and tissues. Glycyrrhetinic acid was used as the internal standard (IS). Chromatographic separation was performed on a 3.5 μm Zorbax SB-C18 column (30 mm × 2.1 mm) with a mobile phase consisting of methanol and aqueous 10 mM ammonium acetate using gradient elution. Quantification was performed by selected ion monitoring with (m/z)⁻ 455 for UA and (m/z)⁻ 469 for the IS. The method was validated in the concentration range of 2.5 − 1470 ng mL⁻¹ for plasma samples and 20 − 11760 ng g⁻¹ for tissue homogenates. The intra- and inter-day assay of precision in plasma and tissues ranged from 1.6% to 7.1% and 3.7% to 9.0%, respectively, and the intra- and inter-day assay accuracy was 84.2 − 106.9% and 82.1 − 108.1%, respectively. Recoveries in plasma and tissues ranged from 83.2% to 106.2%. The limits of detections were 0.5 ng mL⁻¹ or 4.0 ng g⁻¹. The recoveries for all samples were >90%, except for liver, which indicated that ursolic acid may metabolize in liver. The main pharmacokinetic parameters obtained were T _max = 0.42 ± 0.11 h, C _max = 1.10 ± 0.31 μg mL⁻¹, AUC = 1.45 ± 0.21 μg h mL⁻¹ and K _a = 5.64 ± 1.89 h⁻¹. The concentrations of UA in rat lung, spleen, liver, heart, and cerebellum were studied for the first time. This method is validated and could be applicable to the investigation of the pharmacokinetics and tissue distribution of UA in rats.     

Enantioselective piezoelectric quartz crystal sensor for d-methamphetamine based on a molecularly imprinted polymer

A piezoelectric quartz crystal (PQC) sensor based on a molecularly imprinted polymer (MIP) has been developed for enantioselective and quantitative analysis of d-(+)-methamphetamine (d(+)-MA). The sensor was produced by bulk polymerization and the resulting MIP was then coated on the gold electrode of an AT-cut quartz crystal. Conditions such as volume of polymer coating, curing time, type of PQC, baseline solvent, pH, and buffer type were found to affect the sensor response and were therefore optimized. The PQC-MIP gave a stable response to different concentrations of d(+)-MA standard solutions (response time = 10 to 100 s) with good repeatability (RSD = 0.03 to 3.09%; n = 3), good reproducibility (RSD = 3.55%; n = 5), and good reversibility (RSD = 0.36%; n = 3). The linear range of the sensor covered five orders of magnitude of analyte concentration, ranging from 10⁻⁵ to 10⁻¹ μg mL⁻¹, and the limit of detection was calculated as 11.9 pg d(+)-MA mL⁻¹ . The sensor had a highly enantioselective response to d(+)-MA compared with its response to l(−)-MA, racemic MA, and phentermine. The developed sensor was validated by applying it to human urine samples from drug-free individuals spiked with standard d(+)-MA and from a confirmed MA user. Use of the standard addition method (SAM) and samples spiked with d(+)-MA at levels ranging from 1 × 10⁻³ to 1 × 10⁻² μg mL⁻¹ showed recovery was good (95.3 to 110.9%).     

Chemiluminescence determination of chromium(III) and total chromium in water samples using the periodate-lucigenin reaction

A new chemiluminescence (CL) method combined with flow injection technique is described for the determination of Cr(III) and total Cr. It is found that a strong CL signal is generated from the reaction of Cr(III), lucigenin and KIO₄ in alkaline condition. The determination of total Cr is performed by pre-reduction of Cr(VI) to Cr(III) by using H₂SO₃. The CL intensity is linearly related to the concentration of Cr in the range 4.0 × 10⁻¹⁰–1.0 × 10⁻⁶ g mL⁻¹. The detection limit (3s _b) is 1 × 10⁻¹⁰ g mL⁻¹ Cr and the relative standard deviation is 1.9% (5.0 × 10⁻⁸ g mL⁻¹ of Cr(III) solution, n = 11). The method was applied to the determination of Cr(III) and total Cr in water samples and compared satisfactorily with the official method.     

Boronic acid-facilitated α-hydroxy-carboxylate anion transfer at liquid/liquid electrode systems: the EICrev mechanism

The transfer of the α-hydroxy-carboxylates of glycolic, lactic, mandelic and gluconic acid from the aqueous electrolyte phase into an organic 4-(3-phenylpropyl)-pyridine (PPP) phase is studied at a triple-phase boundary electrode system. The tetraphenylporphyrinato complex MnTPP dissolved in PPP is employed to drive the anion transfer reaction and naphthalene-2-boronic acid (NBA) is employed as a facilitator. In the absence of a facilitator, the ability of α-hydroxy-carboxylates to transfer into the organic phase improves, consistent with hydrophobicity considerations giving relative transfer potentials (for aqueous 0.1 M solution) of gluconate>glycolate>lactate>mandelate. In the presence of NBA, a shift of the reversible transfer potential to more negative values is indicating fast reversible binding (the mechanism for the electrode process is EIC_rev) and the binding constants are determined as K _glycolate = 2 M⁻¹, K _mandelate = 60 M⁻¹, K _lactate = 130 M⁻¹ and K _gluconate = 2,000 M⁻¹. The surprisingly strong interaction for gluconate is rationalised based on secondary interactions between the gluconate anion and NBA.     

The Potential Energies of Cohesion of a Crystalline Organic Enantiomer and the Racemate; on the Energy for the Liquid Racemate [1]

The cohesion potential energy of the crystal of one enantiomer of ethyl 3-cyano-3-(3,4-dimethyloxyphenyl)-2,2,4-trimethylpentanoate, −47.7 ± 0.1 kJ mol⁻¹ (0–90°C), was found out from the heat of sublimation (123.2 ± 5.1 kJ mol⁻¹, 78.6°C) and the kinetic energies for the gas phase and the crystal. It was found that the entropy function of Debye’s theory of solids mathematically agreed with the vibrational entropy of the gas (variationally obtained), allowing to disclose the vibrational energy using the Debye energy function (E _vib 835.0 kJ mol⁻¹ (78.6°C), E ⁰ included). E _kin for the crystal (771.1 kJ mol⁻¹ (78.6°C)) was obtained by Debye’s theory with the experimental heat capacity. The cohesion energy represented a moderate part of the sublimation energy. The cohesion energy of the racemic crystal, −44.2 kJ mol⁻¹, was obtained by the heat of formation of the crystal in the solid state (3.0 kJ mol⁻¹, 83.3°C) and E _kin for the crystal (by Debye’s theory). The decrease in cohesion on formation of the crystal accounted for the energy of formation. The change in potential energy on liquefaction of the racemate from the gas state was disclosed obtaining added-up E _{vib + rot} for the liquid in the way as to E _vib for the gas, the Debye entropy function being increasedly suited for the liquid (E _{vib + rot} 763.4 kJ mol⁻¹ (115.4°C)). Positive ΔE _pot, 13.0 kJ mol⁻¹, arised from the increase in electronic energy (Δ _l ν_mean − 154.3 cm⁻¹, by the dielectric nature of the liquid), added to the cohesion energy.     

The Potential Energies of Cohesion of a Crystalline Organic Enantiomer and the Racemate; on the Energy for the Liquid Racemate [1]

Summary. The cohesion potential energy of the crystal of one enantiomer of ethyl 3-cyano-3-(3,4-dimethyloxyphenyl)-2,2,4-trimethylpentanoate, −47.7 ± 0.1 kJ mol⁻¹ (0–90°C), was found out from the heat of sublimation (123.2 ± 5.1 kJ mol⁻¹, 78.6°C) and the kinetic energies for the gas phase and the crystal. It was found that the entropy function of Debye’s theory of solids mathematically agreed with the vibrational entropy of the gas (variationally obtained), allowing to disclose the vibrational energy using the Debye energy function (E _vib 835.0 kJ mol⁻¹ (78.6°C), E ⁰ included). E _kin for the crystal (771.1 kJ mol⁻¹ (78.6°C)) was obtained by Debye’s theory with the experimental heat capacity. The cohesion energy represented a moderate part of the sublimation energy. The cohesion energy of the racemic crystal, −44.2 kJ mol⁻¹, was obtained by the heat of formation of the crystal in the solid state (3.0 kJ mol⁻¹, 83.3°C) and E _kin for the crystal (by Debye’s theory). The decrease in cohesion on formation of the crystal accounted for the energy of formation. The change in potential energy on liquefaction of the racemate from the gas state was disclosed obtaining added-up E _{vib + rot} for the liquid in the way as to E _vib for the gas, the Debye entropy function being increasedly suited for the liquid (E _{vib + rot} 763.4 kJ mol⁻¹ (115.4°C)). Positive ΔE _pot, 13.0 kJ mol⁻¹, arised from the increase in electronic energy (Δ _l ν_mean − 154.3 cm⁻¹, by the dielectric nature of the liquid), added to the cohesion energy.     

Stannum film electrode for square wave voltammetric determination of trace copper(II)

An anodic stripping voltammetric procedure for the determination of Cu(II) at an in situ-plated stannum film electrode (SnFE) was described. The results indicated that the SnFE had an attractive electroanalytical performance, with two distinct voltammetric stripping signals for copper and stannum, and showed the superior advantage for the determination of copper compared with the bismuth film electrode. Several experimental parameters were optimized. The SnFE exhibited highly linear behavior in the concentration range from 1.0 to 100.0 μg L⁻¹ of Cu(II) (r = 0.994) with the detection limit of 0.61 μg L⁻¹ (S/N = 3), and the relative standard deviation for a solution containing 40.0 μg L⁻¹ Cu(II) was 2.2% (n = 8). The procedure has been successfully applied for the determination of Cu(II) in lake water sample.     

Rapid residue analysis of four triazolopyrimidine herbicides in soil, water, and wheat by ultra-performance liquid chromatography coupled to tandem mass spectrometry

A sensitive and effective method for simultaneous determination of triazolopyrimidine sulfonamide herbicide residues in soil, water, and wheat was developed using ultra-performance liquid chromatography coupled with tandem mass spectrometry. The four herbicides (pyroxsulam, flumetsulam, metosulam, and diclosulam) were cleaned up with an off-line C18 SPE cartridge and detected by tandem mass spectrometry using an electrospray ionization source in positive mode (ESI+). The determination of the target compounds was achieved in <2.0 min. The limits of detection were below 1 μg kg⁻¹, while the limits of quantification did not exceed 3 μg kg⁻¹ in different matrices. Quantitation was determined from calibration curves of standards containing 0.05–100 μg L⁻¹ with r ² > 0.997. Recovery studies were conducted at three spiked levels (0.2, 1, and 5 μg kg⁻¹ for water; 5, 10, and 100 μg kg⁻¹ for soil and wheat). The overall average recoveries for this method in water, soil, wheat plants, and seeds at three levels ranged from 75.4% to 106.0%, with relative standard deviations in the range of 2.1–12.5% (n = 5) for all analytes.     

Callus induction and plant regeneration from different explants of Actinidia deliciosa

In this study, an efficient procedure was developed for callus induction and regeneration of kiwifruit (Actinidia deliciosa) using different organs of shoots developed under in vitro conditions. Effects of explants source and media (M₁, 1.0 mg l⁻¹ BA + 2.0 mg l⁻¹ 2,4-D–M₂, 1.0 mg l⁻¹ NAA + 2.0 mg l⁻¹ 2,4-D) on initiation of callus were examined in order to obtain callus for organogenesis. The best callus for plant regeneration was obtained from leaf explants on Murashige and Skoog’s medium (MS) supplemented with M₂. Formation of callus from leaf of kiwifruit (A. deliciosa) was cultured in MS medium containing different concentration of N⁶-benzylaminopurin (BA; 0.0, 1.0, 2.0, 4.0, 6.0, 8.0 mg l⁻¹) for callus proliferation and plant regeneration. Although the first shoot formation was appeared in medium containing 6.0 and 8.0 mg l⁻¹ BA, the best shoots formation was obtained in medium with 4.0 mg l⁻¹ BA.