Study on the supramolecular interaction of thiabendazole and beta-cyclodextrin by spectrophotometry and its analytical application

The beta-cyclodextrin-thiabendazole (beta-CD-TBZ) inclusion complex was synthesized and its structure characterized by (1)H NMR and IR. The mechanism of the supramolecular interaction of TBZ and beta-CD has been studied and discussed by spectrophotometry. The results showed that the phenyl ring of TBZ was included in the beta-CD cavity to form a 1:1 host-guest complex with an apparent formation constant of 1.60 x 10(3) mol(-1).L. On the basis of the enhancement of the absorbance of TBZ produced through complex formation, a spectrophotometric method for the determination of TBZ in bulk aqueous solution in the presence of beta-CD was developed. The linear relationship between the absorbance and TBZ concentration was obtained in the range of 8.86 x 10(-7)-1.45 x 10(-5) mol/L. The detection limit was 2.71 x 10(-7)mol/L, and the relative standard deviation was 0.86%. The interference of 48 coexisting substances was slight. The proposed method has been successfully applied to the determination of TBZ in fruits with recoveries of 96-103%.     

Column preconcentration and spectrophotometric determination of ziram and zineb in commercial samples and foodstuffs using (1,2'-pyridylazo)-2-naphthol (PAN)-naphthalene as adsorbate

A procedure has been developed for the determination of zinc(II) bis(dimethyldithiocarbamate) (ziram) and zinc(II) ethylenebis(dithiocarbamate) (zineb) after preconcentration on a column using naphthalene-(1,2'-pyridylazo)-2-naphthol (PAN) as adsorbent. Ziram and zineb are quantitatively retained on the column in the pH range of 9.0-12.5 and at a flow rate of 1-2 mL/min. The solid mass consisting of the Zn-PAN complex along with naphthalene is dissolved from the column with 5 mL of dimethylformamide (DMF). Absorbance of the complex was measured at 550 nm; Beer's law is obeyed over the concentration ranges of 2.0-22.0 microg of ziram and 5.0-19.8 microg of zineb in 10 mL of the final DMF solution. Ten replicate determinations on a sample solution containing 20 microg of ziram and 18 microg of zineb gave a mean absorbance of 0.33 with relative standard deviations of 0.80 and 0.70%, respectively. The interference of various ions has been studied. The method has been employed for the determination of ziram and zineb in commercial samples and in various foodstuffs, and the results were compared with the earlier reported methods.     

Curcuma longa L. constituents inhibit sortase A and Staphylococcus aureus cell adhesion to fibronectin

The inhibitory activity of Curcuma longa L. (turmeric) rhizome constituents against sortase A, a bacterial surface protein anchoring transpeptidase, from Staphylococcus aureus ATCC 6538p was evaluated. The activity of the isolated compounds (1-4) was compared to that of the positive control,p-hydroxymecuribenzoic acid (pHMB). The biologically active components of C. longa rhizome were characterized by spectroscopic analysis as the curcuminoids curcumin (1), demethoxycurcumin (2), and bisdemethoxycurcumin (3). Curcumin was a potent inhibitor of sortase A, with an IC50 value of 13.8 +/- 0.7 microg/mL. Bisdemethoxycurcumin (IC50 = 31.9 +/- 1.2 microg/mL) and demethoxycurcumin (IC50 = 23.8 +/- 0.6 microg/mL) were more effective than pHMB (IC50 = 40.6 +/- 1.2 microg/mL). The three isolated compounds (1-3) showed no growth inhibitory activity against S. aureus strain Newman, with minimum inhibitory concentrations (MICs) greater than 200 microg/mL. Curcumin also exhibited potent inhibitory activity against S. aureus cell adhesion to fibronectin. The suppression of fibronectin-binding activity by curcumin highlights its potential for the treatment of S. aureus infections via inhibition of sortase activity. These results indicate that curcumin is a possible candidate in the development of a bacterial sortase A inhibitor.     

Development of an extractive spectrophotometric method for the determination of copper(II) in leafy vegetable and pharmaceutical samples using pyridoxal-4-phenyl-3-thiosemicarbazone (PPT)

A highly sensitive extractive spectrophotometric method has been developed for the determination of copper(II) using pyridoxal-4-phenyl-3-thiosemicarbazone(PPT) as an analytical reagent. The PPT forms reddish brown species of copper(II) at a pH range of 3.0-5.5, and the complex was extracted into n-butanol. The Cu(II)-PPT complex shows maximum absorbance at 440 nm, with molar absorptivity and Sandell's sensitivity being 2.16 x 10(4) L mol(-1) cm(-1) and 2.94 x 10(-3) microg cm(-2), respectively. The system obeys Beer's law in the range of 0.2-5.0 mg/L. The regression coefficient of the Beer's law straight line is 0.338, and the correlation coefficient is 0.96. The detection limit of the method is 0.0065 microg mL(-1). Most of the common metal ions generally found associated with copper do not interfere. The repeatability of the method was checked by finding the relative standard deviation. The developed method has been successfully employed for the determination of copper(II) in leafy vegetable and pharmaceutical samples. The method is evaluated by analyzing samples from the Bureau of Analyzed Samples (BCS 233, 266, 216/1, 207, and 179) and by intercomparison of experimental values using AAS.     

Direct spectrophotometric determination of ferbam (Iron(III) dimethyldithiocarbamate) in commercial sample and wheat grains using 4,7-diphenyl-1,10-phenanthroline

A procedure has been developed for the determination of iron(III) dimethyldithiocarbamate by converting it into the iron(II)-bathophenanthroline complex, which is then dissolved in acetone-water (1:1), and the absorbance is measured at 534 nm against a reagent blank. Beer's law is obeyed over the concentration range 0.5-20 microg x mL(-1) in the final solution. The method is sensitive and highly selective and is used for the direct determination of ferbam in a commercial sample and in mixtures with various dithiocarbamates (ziram, zineb, maneb, etc.) and from wheat grains.     

Encapsulation of olive leaf extract in beta-cyclodextrin

Olive leaf extract, rich in oleuropein, formed an inclusion complex with beta-cyclodextrin (beta-CD) upon mixing of the components in aqueous media and subsequent freeze-drying. Inclusion complex formation was confirmed by differential scanning calorimetry (DSC). DSC thermograms indicated that the endothermic peaks of both the olive leaf extract and the physical mixture of olive leaf extract with beta-CD, attributed to the melting of crystals of the extract, were absent in DSC thermogram of inclusion complex. Moreover, DSC studies under oxidative conditions indicated that the complex of olive leaf extract with beta-CD was protected against oxidation, since it remained intact at temperatures where the free olive leaf extract was oxidized. Phase solubility studies afforded A L type diagrams, 1:1 complex stoichiometry, a moderate binding constant ( approximately 300 M (-1)), and an increase of the aqueous solubility by approximately 50%. The formation of the inclusion complex was also confirmed by nuclear magnetic resonance (NMR) studies of beta-CD solutions in the presence of both pure oleuropein and olive leaf extract. The NMR data have established the formation of a 1:1 complex with beta-CD that involves deep insertion of the dihydroxyphenethyl moiety inside the cavity from its secondary side.     

Sensitive determination of sulfate in drinks and vegetables digests by Rayleigh light scattering technique

In this paper, a novel and sensitive method based on Rayleigh light scattering technique (RLS) was proposed for the determination of sulfate using a conventional spectrofluorometer. Sulfate was transformed to BaSO4 particles, which displayed intense light scattering in aqueous solutions. The effects of factors such as wavelength, acidity, stabilizers and interferents were studied in detail. The RLS intensity of the BaSO4 suspension was obtained in 0.1 mol L(-1) of [H+] and the addition of 2 mL of cationic polyacrylamide (CPAM) with 7.05 x 10(-3) mmol mL(-1) charged cations and 1.0 mL of BaCl2.2H2O (5.0%) at 510 nm. In the range of 8-400 microg mL(-1), RLS intensity was linear to the concentration of BaSO4, and the detection limit was 0.3 microg mL(-1). To determine the feasibility of the proposed method, some samples of water, drinks, and vegetables digests were analyzed, and the results were in agreement with the standard turbidimetric method. Good recovery results were also obtained in the range of 94-105%. Although this method was limited in stability, it was characterized with simplicity, sensitivity, reliability, and little interference.     

Green-leaf-derived C6-aroma compounds with potent antibacterial action that act on both Gram-negative and Gram-positive bacteria

All eight C6-aliphatic alcohol and aldehyde compounds in naturally occurring green leaves showed bacteriostatic effects against Staphylococcus aureus IFO 12732, methicillin-resistant S. aureus, Escherichia coli IFO 3301, E. coli O157:H7, and Salmonella enteritidis, with bacteriostatic activities of less than 12.5 microg mL(-1). In this study, the susceptibility of Gram-positive bacteria tested was observed to be greater than that of Gram-negative bacteria. The bactericidal action of the aldehyde compounds was found to be much stronger than that of the alcohol compounds under both liquid and gaseous conditions. The most effective compound was (3E)-hexenal at concentrations of 0.1 and 1 microg mL(-1), which killed 2.1 x 10(5) cfu mL(-1) of S. aureus IFO 12732 and 1.4 x 10(5) cfu mL(-1) of E. coli IFO 3301, respectively, by direct contact with the compound. Lethality of (3E)-hexenal against S. aureus IFO 12732 and E. coli IFO 3301 was also observed as a result of gaseous contact at concentrations of 3 and 30 microg mL(-1), respectively. The bactericidal effects of 30 microg mL(-1) (3E)-hexenal were thoroughly maintained throughout periods of 2 days and 1 day against S. aureus IFO 12732 and E. coli IFO 3301, respectively, by a complex formation with alpha-cyclodextrin.     

Thermal stability of anthocyanin extract of Hibiscus sabdariffa L. in the presence of beta-cyclodextrin

The thermal stability of anthocyanin extract isolated from the dry calyces of Hibiscus sabdariffa L. was studied over the temperature range 60-90 degrees C in aqueous solutions in the presence or absence of beta-cyclodextrin (beta-CD). The results indicated that the thermal degradation of anthocyanins followed first-order reaction kinetics. The temperature-dependent degradation was adequately modeled by the Arrhenius equation, and the activation energy for the degradation of H. sabdariffa L. anthocyanins during heating was found to be approximately 54 kJ/mol. In the presence of beta-CD, anthocyanins degraded at a decreased rate, evidently due to their complexation with beta-CD, having the same activation energy. The formation of complexes in solution was confirmed by nuclear magnetic resonance studies of beta-CD solutions in the presence of the extract. Moreover, differential scanning calorimetry revealed that the inclusion complex of H. sabdariffa L. extract with beta-CD in the solid state was more stable against oxidation as compared to the free extract, as the complex remained intact at temperatures 100-250 degrees C where the free extract was oxidized. The results obtained clearly indicated that the presence of beta-CD improved the thermal stability of nutraceutical antioxidants present in H. sabdariffa L. extract, both in solution and in solid state.     

Analysis of tea shoot catechins: spectrophotometric quantitation and selective visualization on two-dimensional paper chromatograms using diazotized sulfanilamide.

A highly specific and sensitive diazotized sulfanilamide reagent is synthesized for determination of tea catechins. The reagent is employed both as spray reagent for selective visualization of tea catechins on two-dimensional paper chromatograms (sensitivity <1 microg of d-(+)-catechin) and for their spectrophotometric quantification in the crude extracts of tea polyphenols isolated from fresh or dried tea shoots. The formation of yellow color (lambda(max) = 425 nm) between catechins and diazotized sulfanilamide was investigated and made the basis of a simple and sensitive spectrophotometric method for estimation of the total and individual catechins in different tea cultivars. At 425 nm, the absorbance was linear (r = 0.999) over the (0.4-8.0 microg/mL) concentration range of d-(+)-catechin.     

Preparation and characterization of inclusion complex of iprodione and beta-cyclodextrin to improve fungicidal activity

The effect of beta-cyclodextrin (beta-CD) on the improvement of the fungicidal activity of iprodione has been investigated. The inclusion complexation of beta-CD with iprodione has been prepared and characterized by integrating some analytical techniques (such as electrospray ionization-mass spectrometry, differential scanning calorimetry, thermogravimetry, x-ray diffraction, and scanning electron microscopy) and molecular simulation methods. The beta-CD/iprodione inclusion complex has exhibited different spectroscopic features and properties from iprodione. The stoichiometric ratio and stability constant describing the extent of formation of inclusion complexes have been determined by phase solubility studies. The calculated apparent stability constant of the iprodione/beta-CD complex was 407.5 M-1. The obtained inclusion complexes were found to significantly improve the water solubility of iprodione, and there is a 4.7-fold increase in the presence of 13 mM beta-CD as compared with the solubility of iprodione in deionized water in the absence of beta-CD. The bioassay demonstrated that the complex displayed over two-fold increase of the fungicidal activity. In addition, the possible structure of the beta-CD/iprodione complex was proposed according to the results of the molecular dynamic simulation. The present study provided useful information for a more rational application of iprodione, diminishing the use of organic solvents and increasing its efficacy.     

Spectrophotometric determination of ferbam [iron(III) dimethyl dithiocarbamate] in commercial sample and wheat grains after extraction of its bathophenanthroline tetraphenylborate complex into molten naphthalene

A procedure has been developed for the determination of iron(III) dimethyldithiocarbamate by converting it into a iron(III)-bathophenanthroline-tetraphenylborate complex, which was then extracted into molten naphthalene, and the absorbance was measured at 534 nm against a reagent blank. Beer's law is obeyed over the concentration range 0.4-20 microg mL(-)(1) in final solution. The method is sensitive and highly selective and is applied for the determination of ferbam in a commercial sample, in mixtures with various dithiocarbamates (ziram, zineb, maneb, etc.), and from wheat grains.     

Preparation and characterization of inclusion complex of norflurazon and beta-cyclodextrin to improve herbicide formulations

The formulation of inclusion complexes of the herbicide norflurazon as guest and beta-cyclodextrin (beta-CD) as host has been studied as a first step in the use of cyclodextrins to obtain improved formulations of this herbicide. The interaction of norflurazon with beta-CD produced the formation of an inclusion complex in solution and in solid state. The inclusion of norflurazon in beta-CD in solution was studied by phase solubility, and an apparent stability constant of 360 M(-)(1), a 1:1 stoichiometric ratio for the complex, and up to 5-fold increase in norflurazon solubility were determined. Three processing methods (kneading, spray drying and vacuum evaporation) were used to prepare norflurazon-beta-CD solid inclusion complexes. X-ray diffraction, infrared spectroscopy, differential scanning calorimetry, and scanning electron microscopy techniques were used to study the solid complexes. From the different solid systems, an increase of norflurazon aqueous dissolution rate was obtained in comparison to the uncomplexed herbicide. This finding is a first step to obtain controlled release and/or protective formulations of norflurazon, which allow a more rational application of norflurazon, diminishing the use of organic solvents and increasing its efficacy.     

Crocetin, dimethylcrocetin, and safranal bind human serum albumin: stability and antioxidative properties

Crocetin (CRT) and dimethylcrocetin (DMCRT) are derived from crocins which are found in the stigmas of saffron (Crocus sativus L.), while safranal is the main component of saffron's essential oil. The aim of the present study was to examine their interaction with human serum albumin in aqueous solution at physiological conditions using constant protein concentration and various ligand contents. FT-IR and UV-visible spectroscopic methods were used to determine the ligands' binding mode, the binding constant, and the effects of ligand complexation on protein secondary structure. Structural analysis showed that crocetin, dimethylcrocetin, and safranal bind nonspecifically (H-bonding) via protein polar groups with binding constants of Kcrt =2.05 (+/-0.30) x 103 M-1, Kdmcrt = 9.60 (+/-0.35) x 104 M-1, and Ksaf = 2.11 (+/-0.35) x 103 M-1. The protein secondary structure showed no major alterations at low ligand concentrations (1 microM), whereas at higher content (1 mM), decrease of alpha-helix from 55% (free HSA) to 43-45% and increase of beta-sheet from 17% (free HSA) to 18-22% and random coil 7% (free HSA) to 10-14% occurred in the ligand-HSA complexes. The results point to a partial unfolding of protein secondary structure at high ligand content. The antioxidant activity of CRT, DMCRT, and safranal was also tested by the DPPH* antioxidant activity assay, and their IC50 values were compared to that of well-known antioxidants such as Trolox and butylated hydroxy toluene (BHT). The IC50 values of CRT and safranal were 17.8 +/- 1 microg/mL and 95 +/- 1 microg/mL, respectively, while the inhibition of DMCRT reached a point of 38.8%, which corresponds to a concentration of 40 microg/mL, and then started to decrease. The IC50 values of Trolox and BHT were 5.2 +/- 1 microg/mL and 5.3 +/- 1 microg/mL, respectively.     

Quantification of uronic acids in tea polysaccharide conjugates and their antioxidant properties

A technique of high-performance liquid chromatography (HPLC) was described for the measurement of total uronic acids in tea polysaccharide conjugates. This method was applied to polysaccharide conjugate extracts obtained from green tea after most of the components that produce interference were removed. The preliminary extraction process was according to the procedure of isolation of polysaccharide conjugates. The uronic acid content of different polysaccharide conjugate fractions was quantified by HPLC on a Sugar-Pak I column with a 1.0 x 10(-)(4) mol x L(-)(1) calcium disodium ethylenediaminetetraacetic acid solution as the mobile phase and refractive index detection. The validation study showed high recoveries (>97.0%) and low coefficients of variance (<3.0%). The minimum detectable limit concentration of uronic acid was 10 microg x mL(-)(1). The analysis of a standard range of galacturonic acid concentrations (100-4000 microg x mL(-)(1)) yielded linear results. The use of the method on different polysaccharide conjugate fraction samples confirmed its effectiveness. With the high content of uronic acids in polysaccharide conjugates, the stronger reactive oxygen species scavenging activities were found.     

Sulfluramid volatility reduction by beta-cyclodextrin

Sulfluramid is an expensive active principle of insecticidal baits that is lost by volatilization during the pelletization of baits. To increase the thermal stability of sulfluramid, we tested its molecular encapsulation in beta-cyclodextrin (beta-CD), using molar ratios of 1:1 and 1:2 (sulfluramid:beta-CD), using the complex preparation techniques of coprecipitation and kneading. The physical mixture of sulfluramid and beta-CD was also tested for comparison. The products of complexation were characterized by differential scanning calorimetry, thermogravimetry, and derivative thermogravimetry, indicating the formation of a sulfluramid/beta-CD complex and showing that the release of the complexed sulfluramid occurs in the range of 270-300 degrees C, a temperature range that is well above the temperature at which sulfluramid sublimates (40 degrees C). This result warrants a reduced sulfluramid loss in the preparation of insecticidal baits. The preparation of the complex by kneading with molar ratio of 1:2 gave the highest yield of complex, about 64%, in relation to the theoretical maximum.     

Improvement of the desorption of the herbicide norflurazon from soils via complexation with beta-cyclodextrin

The effect of beta-cyclodextrin (beta-CD) on the removal of the herbicide norflurazon (NFL) from soils has been investigated. The interaction of NFL with beta-CD in solution yielded the formation of a water-soluble inclusion complex at 1:1 stoichiometric ratio, which gave an increase in NFL solubility. Desorption studies of NFL previously adsorbed on six soils of different characteristics have been performed in the presence of 0.01 M beta-CD or 0.01 M Ca(NO(3))(2) as extractant solutions. Positive hysteresis was observed in all soils when 0.01 M Ca(NO(3))(2) solution was used, indicating that desorption of NFL from these soils was not completely reversible. On the contrary, the application of beta-CD solutions to soils where NFL had been previously adsorbed increased very much its desorption, and a negative hysteresis was obtained for all soils studied; that is, more NFL was desorbed with respect to NFL adsorption isotherm. A clear relationship was observed between the physicochemical characteristics of the soils and the beta-CD concentration necessary to remove the herbicide, the percentages of desorption observed for each soil being inversely related to the values obtained for the Freundlich sorption capacity parameter of the herbicide, K(f). In general, high desorption yields can be obtained with very low beta-CD concentrations, which is an important advantage from an economic point of view, although in those soils that present an extremely high NFL adsorption, higher amounts of beta-CD should be used. The results obtained indicate the high extracting power of beta-CD toward the herbicide previously adsorbed on the soils and the potential use of beta-CD for in situ remediation of pesticide-contaminated soils.     

Development of an improved liquid phase microextraction technique and its application in the analysis of flumetsulam and its two analogous herbicides in soil

An improved liquid phase microextraction (LPME) technique has been developed. As part of this technique, analytes were extracted into an extractant microdrop which was laid on the cone-shaped bottom of a PCR tube (polychloroprene rubber tube) but not at the needle tip of a microsyringe, and the sample vial and PCR tube were horizontally placed so that the extractant was not affected by the force of vertical orientation (gravity and floating force). The stability of the extractant microdrop increased greatly, and the selection of extractant was extended. In this work, flumetsulam and its two analogous herbicides were chosen as model analytes in investigating the feasibility of the new pretreatment method by coupling it to high-performance liquid chromatography (HPLC). Under the optimized experimental conditions, the linear range and the limits of detection (S/N = 3) were 0.01-5 microg/mL (r = 0.9997) and 0.8 ng/mL for flumetsulam, 0.002-5 microg/mL (r = 0.9994) and 0.5 ng/mL for analogue 1, and 0.002-1 microg/mL (r = 0.9993) and 0.5 ng/mL for analog 2, respectively. The inter- and intraday reproducibilities (RSD) were below 5.3 and 4.5%, respectively. Good recoveries that ranged from 79.4 to 115.0% were obtained in the analysis of real soil samples. The extraction efficiency of the improved method was 4-8 times higher than that of the conventional liquid phase microextraction method. The novel, simple, rapid, sensitive technique is very suitable for extraction of apolar and medium polar analyte in complex environmental samples.     

Rapid extraction and high-performance liquid chromatographic determination of parthenolide in feverfew (Tanacetum parthenium).

A rapid and sensitive method for quantifying parthenolide in feverfew herb (Tanacetum parthenium) was developed that is significantly faster than those reported in the literature. The extraction system consisted of acetonitrile/water (90:10, v/v) in a bottle with stirring for 30 min. Both Soxhlet and bottle-stirring extractions were studied. Samples were analyzed using high-performance liquid chromatography with a Cosmosil C18-AR column (150 x 4.6 mm, 5 microm, 120 A). The mobile phase consisted of acetonitrile/water (55:45, v/v) with a flow rate of 1.5 mL/min and UV detection at 210 nm. Analysis time was 6 min, with a detection limit of 0.10 ng on column. The calibration curve was linear over a range of 0.160-850 microg/mL parthenolide with R(2) = 0.9999. Replicate tests indicated good reproducibility of the method with an RSD% = 0.88 (n = 10). Spike recovery of parthenolide was found to be 99.3% with an RSD% = 1.6 (n = 6).     

Spectrophotometric determination of tolbutamide in tablets.

Two spectrophotometric methods, Glenn's method of orthogonal function and the basic dye method, are described for determing tolbutamide in tablets without interference from the tablet excipients. In Glenn's method, the absorbance of tolbutamide in 95% ethanol is measured in the vicinity of 250--270 nm at 4 nm intervals and the p2 coefficient is calculated. The coefficient is linearly related to concentration within a range of 0.1--0.4 mg/mL. Tolbutamide gives a complex of a ratio 1:1 with basic dye Brilliant Cresyl Blue (BCB) or Safranin T (ST). The complex is easily extracted with chloroform. The absorbance of the chloroform extract is measured against either a blank or reference experiment. The latter is obtained by using a specific concentration of tolbutamide: 0.4 mg/mL in tolbutamide-BCB or 0.024 mg/mL in tolbutamide-ST. The ST complex method is more sensitive compared with the other methods. When the t-test is applied, the results of the proposed methods are more accurate than those of the traditional ultraviolet spectrophotometric method.