Kinetic study of the tocopherol regeneration reaction by biological hydroquinones in micellar solution

The rate constants (kr) of the regeneration reaction of 7-t-butyl-5-isopropyltocopheroxyl with ubiquinol-10 (UQ10H2), ubiquinol-0 (UQ0H2), alpha-, beta-, and gamma-tocopherolhydroquinones (alpha-, beta-, gamma-TQH2), 2,3,5-trimethyl-1,4-hydroquinone (TMQH2), vitamin K3 hydroquinone (VK3H2), and vitamin C (Vit C) have been measured in 2-propanol/water and micellar solutions by a stopped-flow spectrophotometer. The kr values of these hydroquinones (HQs) in micellar solution remained constant at pHs of 6-9 and increased rapidly by increasing the pH value. The kr values decreased in the order of VK3H2 > gamma-TQH2 > or = alpha-TQH2 > beta-TQH2 > or = UQ10H2 > or = TMQH2 > UQ0H2 > Vit C at pHs of 6-9. These HQs are dibasic acids and can exist in three different molecular forms, depending on pH. By comparing the kr values with the mole fraction of each molecular form of the HQs, the reaction rate kr1 for the undissociated form, kr2 for the monoanion, and kr3 for the dianion and the pKa1 and pKa2 values were determined. It has been found that the kr values of UQ10H2, alpha-TQH2, beta-TQH2, and gamma-TQH2 (plastoquinol model) are 460, 1430, 494, and 1530 times larger than that of Vit C at pH 7.0, respectively, although the values are similar to that of Vit C in 2-propanol/water. The biological HQs and Vit C coexist in many tissues of animals and plants, and thus, the relative antioxidant activities of HQs and Vit C have been tentatively discussed based on the products of kr values by concentrations in several tissues. The results suggest that these HQs show high activity for the tocopherol regeneration in biological systems.     

Normal-phase high-performance liquid chromatography of tocopherols and tocotrienols. Comparison of different chromatographic columns

Natural vitamin E is composed of eight different vitamers (alpha-, beta-, gamma- and delta-tocopherols and alpha-, beta-, gamma- and delta-tocotrienols). As these eight vitamers have different antioxidant and biological activities, it is necessary to have quantitative data on each substance separately. The aim of this study was to find universal HPLC columns for the separation of all eight components and to test if a few columns of the same material (different batches) will give reproducible results. Normal-phase HPLC separations of vitamin E compounds in a prepared mixture (containing oat extracts, palm oil and tocopherol standards) were tried on six silica, three amino and one diol columns. As shown by calculations of retention factors (k), separation factors (alpha), numbers of theoretical plates (N) and resolutions (Rs), the best separations were obtained on three silica columns and two amino columns using 4 or 5% dioxane in hexane as the mobile phase as well as on a diol column using 4% tert.-butyl methyl ether in hexane as the mobile phase.     

Liquid chromatographic method for the analysis of tocopherols in malt sprouts with supercritical fluid extraction.

A simple, specific and sensitive high-performance liquid chromatographic method has been developed for the determination of tocopherols in malt sprouts. A supercritical fluid extraction (SFE) procedure was used to isolate tocopherols from the vegetal matrix before quantitative analysis. The analytes were separated on a Zorbax reversed-phase column using methanol-water as mobile phase and quantified by measuring its fluorescence at lambda(em)=328 nm after excitation of the analytes at lambda(exc)=303 nm. The limits of detection for alpha-, gamma- and delta-tocopherols were 0.04, 0.05, and 0.05 microg/ml, respectively. The calibration graphs of the method were linear from 0.1 to 1.5, 0.2 to 2.5, and 0.2 to 2.0 microg/ml, for alpha-, gamma- and delta-tocopherols, respectively. This SFE and HPLC procedure is simple, precise and accurate for the determination of tocopherols in malt sprouts.     

Stability and structure in alpha- and beta-Keggin Heteropolytungstates, [X(n)(+)W12O40)](8-n)(-), X = p-block cation

beta-[SiW(12)O(40)](4)(-) (C(3)(v) symmetry) is sufficiently higher in energy than its alpha-isomer analogue that effectively complete conversion to alpha-[SiW(12)O(40)](4)(-) (T(d)) is observed. By contrast, beta- and alpha-[AlW(12)O(40)](5)(-) (beta- and alpha-1; C(3)(v) and T(d), respectively) are sufficiently close in energy that both isomers are readily seen in (27)Al NMR spectra of equilibrated (alpha-beta) mixtures. Recently published DFT calculations ascribe the stability of beta-1 to an electronic effect of the large, electron-donating [AlO(4)](5)(-) (T(d)) moiety encapsulated within the polarizable, fixed-diameter beta-W(12)O(36) (C(3)(v)) shell. Hence, no unique structural distortion of beta-1 is needed or invoked to explain its unprecedented stability. The results of these DFT calculations are confirmed by detailed comparison of the X-ray crystal structure of beta-1 (beta-Cs(4.5)K(0.5)[Al(III)W(12)O(40)].7.5H(2)O; orthorhombic, space group Pmc2(1), a = 16.0441(10) A, b = 13.2270(8) A, c = 20.5919(13) A, Z = 4 (T = 100(2) K)) with previously reported structures of alpha-1, alpha- and beta-[SiW(12)O(40)](4)(-), and beta(1)-[SiMoW(11)O(40)](4)(-).     

Mechanism of quinol oxidation by ferricenium produced by light excitation in reaction centers of photosynthetic bacteria

The kinetics and thermodynamics of cyclic electron transfer through the isolated reaction center protein of photosynthetic bacterium Rhodobacter sphaeroides were determined in detergent (Triton X-100) solution. The redox reactions between the reducing (ubiquinol-0 or ubiquinol-10) and oxidizing species (ferricenium, ferricytochrome, or ferricyanide) produced chemically or by light excitation of the protein were monitored by absorption changes of the reactants and by acidification of the solution accompanied with the disappearance of the quinol. The bimolecular rate constants of reactions of anionic ubiquinol-0 with different oxidizing agents showed large variation: 5 x 10(8) M(-1) s(-1) for ferricenium, 3.5 x 10(5) M(-1) s(-1) for ferricyanide, and 1.5 x 10(5) M(-1) s(-1) for ferricytochrome. Although the redox partners were created in pairs by the same protein promptly after light excitation, their bimolecular redox reaction was not observed even in the case of the fastest reacting partners of ferricenium and ubiquinol-0. Instead, they equilibrate with the corresponding (donor and acceptor) pools before the electron is transferred. The (logarithms of the) observed rate constants of quinol oxidation showed steep pH-dependence for water soluble ubiquinol-0 (slope +1) and mild pH-dependence for hydrophobic ubiquinol-10 (slope approximately 0.25). Combined with studies of the ionic strength dependence of the rate, it was concluded that the electron-transfer pathways of ubiquinol-0 and ubiquinol-10 oxidation started from their anionic and neutral forms, respectively. The mild pH-dependence of the rate of ubiquinol-10 oxidation came from the electrostatic interactions between ferricenium and the pH-dependent surface charges of the reaction center. The results help to understand, monitor, and design (cyclic) electron flow in bioenergetic proteins.     

Simple reversed-phase liquid chromatography method for determination of tocopherols in edible plant oils

A simple and rapid reversed-phase high-performance liquid chromatography method for determination of alpha-, (beta + gamma), and delta-tocopherols in edible plant oils has been developed. Oils are diluted in 2-propanol and injected directly onto Symmetry C18 column. Methanol and acetonitrile (1:1) are used as a mobile phase. Tocopherols are detected using fluorescence detector set at excitation and emission wavelength 295 nm and 325 nm, respectively. The method is precise (R.S.D. not higher than 2.24%) and sensitive-detection limits (DL) are 8 ng/ml for gamma- and delta-tocopherols and 28 ng/ml for alpha-tocopherol; quantification limits (QL) were calculated as three times higher than DL.     

Trace analysis of heavy metals in groundwater samples by ion chromatography with post-column reaction and ultraviolet-visible detection

A rapid and automated method for the analysis of alpha-, gamma- and delta-tocopherols in vegetable oils is reported. Continuous extraction of vitamin E isomers from oil samples dissolved in Triton X-114 in the presence of methanol-hexane is achieved and coupled on-line with the chromatographic system. Using an acetic acid/sodium acetate buffer in a methanol-water (97:3) solution as the mobile phase, a C18 stationary phase and electrochemical detection in the coulometric mode, alpha-, gamma- and delta-tocopherol isomers can be successfully analyzed within 17 min. Thirteen commercially available oils of olive, sunflower, corn and seed mixtures were analyzed using 2,2,5,7,8-pentamethyl-6-chromanol as internal standard. The results obtained using three methodologies, one of them including classical sample treatment for liposoluble vitamin analysis, were in good agreement. To validate the proposed method, analysis of the only BCR Reference Material available, with a certified content of alpha-tocopherol (margarine CRM 122), was carried out using the automated methodology, the results found being in agreement with the certified value.     

Use of a polar-embedded stationary phase for the separation of tocopherols by CEC

A polar-embedded stationary phase (ULTIMA C18) has been investigated for the separation of alpha-, beta-, gamma- and delta-tocopherols by CEC in comparison with commercially available C(18) and C(30) n-alkyl RPs. The behavior of this stationary phase was tested for different mobile phases based on methanol, ACN, or mixtures thereof and different separation parameters such as retention factors and resolution were evaluated. The main feature of this stationary phase is the improved selectivity for the separation of beta- and gamma-tocopherols (positional isomers) when compared with the pure n-alkyl C(18) material, which was unable to resolve these compounds. Additionally, it is possible to observe a reversal in the elution order of the beta- and gamma-tocopherol isomers with respect to that obtained on the C(30) column. The resulting data indicate that the enhanced selectivity obtained with the polar-embedded stationary phase, with respect to the conventional C(18) material, is due to the participation of both hydrophobic and polar interactions: these latter are of the hydrogen bridge type with the amide group of the polar-embedded stationary phase, which increases the retention of the tocopherols and facilitates the discrimination between the beta- and gamma-isomers. Adequate separation of the four tocopherols was obtained by CEC using the polar-embedded stationary phase and 95:5 v/v methanol/water (5 mM Tris, final concentration) as the mobile phase.     

A common carbanion intermediate in the recombination and proton-catalysed disproportionation of the carboxyl radical anion, CO2*-, in aqueous solution

The carboxyl radical anion, CO2*- was produced by the reactions of OH radicals with either CO or formic acid in aqueous solution. The pKa(*CO2H) was determined by pulse radiolysis with conductometric detection at pH approximately equals 2.3. The bimolecular decay rate constant of CO2*- (2k approximately equals 1.4 x 10(9) dm3mol(-1)s(-1)) was found to be independent of pH in the range 3-8 at constant ionic strength. The yields of the products of the bimolecular decay of the carboxyl radicals, CO2 and the oxalate anion were found to depend strongly on the pH of the solution with an inflection point at pH 3.8. This pH dependence is explained by assuming a head-to-tail recombination of the CO2*- radicals followed by either rearrangement to oxalate or a protonation of the adduct, which subsequently leads to the formation of CO2 and formate. The recombination of CO2*- to give oxalate directly is estimated to have a contribution of <25%.     

Comparison of the column performance of narrow-bore and standard-bore columns for the chromatographic determination of alpha-, beta-, gamma-, and delta-tocopherol

A comparison of the performance of narrow-bore (2.1-mm i.d.) and standard-bore (4.6-mm i.d.) analytical silica columns having the same length is completed for the resolution of alpha-, beta-, gamma-, and delta-tocopherol. The studies are performed on high-performance liquid chromatographic equipment with minimum extracolumn contribution. Column permeabilities are 1.16 x 10(-9) and 2.48 x 10(-9) cm2 for narrow and standard bore, respectively. The narrow-bore column gives up to a 7 times increase in sensitivity compared with a standard-bore column at equivalent running times for the analytes. Approximately one-third solvent savings can be achieved with the narrow-bore column. Theoretical plates of the standard-bore column are higher than that of the narrow-bore column.     

Oxidation of phenol in aqueous acid: characterization and reactions of radical cations vis-à-vis the phenoxyl radical

Aqueous sulfuric acid containing up to approximately 14 M acid (H0 > or = -7.0) was used as solvent in pulse radiolytic redox studies to characterize cationic transients of phenol (C6H5OH) and map their reactions. The primary radical yields were first measured to correlate the variation in various radical concentrations as a function of increasing acid fraction in the solvent. Compared to their respective values at pH 2, the G(Ox*) increased with almost a linear slope of approximately 0.024 micromol J(-1) for H0(-1) (or pH(-1)) up to H0 -6.0 (Ox* = *OH + SO4*-), whereas G(H*) increased with a slope of approximately 0.033 micromol J(-1) for H0(-1) (or pH(-1)) up to H0 -5.0. In the presence of > 10 M acid (H0 < -5.0), phenol was oxidized to its radical cation, C6H5OH*+, which further reacted with phenol and generated the secondary, dimeric radical cation, (C6H5OH)2*+, following an equilibrium reaction C6H5OH*+ + C6H5OH <==> (C6H5OH)2*+, with K(eq) = 315 +/- 15 M(-1). The two cationic radicals were characterized from their individual UV-vis absorption spectra and acidity. The C6H5OH*+ absorption peaks are centered at 276 and 419 nm, and it was found to be more acidic (pKa = -2.75 +/- 0.05) than (C6H5OH)2*+ (pKa = -1.98 +/- 0.02), having its major peak at 410 nm. On the other hand, in the presence of < 6.5 M acid the C6H5O* reactions followed the radical dimerization route, independent of the parent phenol concentration.     

Three conformational polymorphs of di-mu-chlorotetrakis(1-methylboratabenzene)diyttrium: synthesis, x-ray structures, quantum chemical calculations, and lattice energy minimizations

The reaction of yttrium trichloride with lithium 1-methylboratabenzene (1/2) in toluene (110 degrees C, 3 days) afforded the donor-free dinuclear sandwich complex [(C(5)H(5)BMe)(2)Y(mu-Cl)](2) (1) in 85% yield as pale-yellow crystals. By means of single crystal and powder diffraction methods, three conformational polymorphs, alpha-1 [P2(1)/n (No. 14), monoclinic, a = 6.6124(8) A, b = 14.352(9) A, c = 14.120(1) A, beta = 95.57(1) degrees, V = 1333.7(9) A(3), Z = 2], beta-1 [P2(1)/a (No. 14), monoclinic, a = 8.542(2) A, b = 13.712(6) A, c = 11.76(1) A, beta = 102.60(4) degrees, V = 1344.5(13) A(3), Z = 2], and gamma-1 [Pbca (No. 61), orthorhombic, a = 20.091(5) A, b = 13.527(3) A, c = 9.976(2) A, V = 2711.2(11) A(3), Z = 4], were characterized in the solid state of 1. The molecules in the three phases vary remarkably in the rotational position of boratabenzene ligands with differences of 91.1, 133.1, and 24.9 degrees between each pair. DFT calculations at the B3LYP/LanL2DZ level reveal that the three molecular structures observed in the solid state correspond closely to three minima on the gas-phase potential energy surface. The beta conformation is 2.8 and 7.2 kJ/mol more stable than the alpha and gamma conformations, respectively. Lattice energy minimizations predict that the alpha-1 phase is about 5.5 and 18.7 kJ mol(-)(1) more stable than the beta-1 and gamma-1 modifications, in agreement with the packing coefficients and the molecular volumes of the three crystal structures. While the alpha-1 and beta-1 modifications have comparable total energies, the gamma-1 form is less stable. The total energy differences among the polymorphs are greater than generally expected.     

Separation of alpha-, beta-, gamma-, delta-tocopherols and alpha-tocopherol acetate on a pentaerythritol diacrylate monostearate-ethylene dimethacrylate monolith by capillary electrochromatography

This work reports the first use of a monolith with method development for the separation of tocopherol (TOH) compounds by CEC with UV detection. A pentaerythritol diacrylate monostearate-ethylene dimethacrylate (PEDAS-EDMA) monolithic column has been investigated for an optimised condition to separate alpha-, beta-, gamma- and delta-TOHs, and alpha-tocopherol acetate (TAc). The PEDAS-EDMA monolith showed a remarkably good selectivity for separation of the TOH isomers including the beta- and gamma-isomers which are not easily separated by standard C8 or C18 particle-packed columns. Retention studies indicated that an RP mechanism was involved in the separation on the PEDAS-EDMA column, but polar interactions with the underlying ester and hydroxyl groups enhanced the separation of the problematic beta- and gamma-isomers. Separation of all the compounds was achieved within 25 min using 3:10:87 v/v/v 100 mM Tris buffer (pH 9.3)/methanol/ACN as the mobile phase. The method was successfully applied to a pharmaceutical sample with recoveries from 93 to 99%. Intraday and interday precisions (%RSD) for peak area and retention time were less than 2.3. LODs for all four TOHs and TAc were below 1 ppm.     

The supertristeroids: large, chiral, molecular bowls prepared by trimerization of pentacyclic steroidal ketones

Robinson annulation of coprostanone (1) at the 2,3- and 3,4-positions gave two pentacyclic enones (7 and 10) that contain A/B-cis-fused ring junctions. Reduction of these enones gave the pentacyclic steroidal ketones 2 alpha,3beta- (8) and 2 alpha,3 alpha-(3'-oxocyclohexano)-5 beta-cholestane (9) and 4 alpha,3beta- (11) and 4 alpha,3 alpha-(3'-oxocyclohexano)-5 beta-cholestane (12). The structures of compounds 8, 9, and 11 were unambiguously established by X-ray analysis. TiCl4-promoted trimerization of compounds 8 and 11 gave the "supertristeroids" 4 and 5, respectively: large (C93) chiral, hydrocarbon clefts with C3-symmetric pockets approximately 12 A in diameter.     

Significant differences in the electrochemical behavior of the alpha-, beta-, gamma-, and delta-tocopherols (vitamin E)

Alpha-, beta-, gamma-, and delta-tocopherols can be oxidized in dry CH2Cl2 or CH3CN by one electron to form cation radicals that deprotonate to form the neutral phenoxyl radicals, which are then immediately further oxidized by one electron to the phenoxonium cations (an ECE electrochemical mechanism, where E signifies an electron transfer and C represents a chemical step, with the electrochemical mechanism having been determined by in situ spectroscopic analysis). The principal difference in the electrochemical behavior of the tocopherols relates to the stability of their associated phenoxonium cations. The phenoxonium cation of alpha-tocopherol is stable in solution for at least several hours, the phenoxonium cation of beta-tocopherol is stable for several minutes, and the phenoxonium cations of gamma- and delta-tocopherol are stable for <1 s. In dry CH2Cl2 containing >0.75 M acid (CF3COOH), the deprotonation reaction of the cation radicals can be completely inhibited resulting in the cyclic voltammetric behavior of the tocopherols appearing as chemically reversible one-electron oxidation processes (an E mechanism). In dry acid conditions, the cation radicals can be further oxidized by one electron to form the dications, which are unstable and immediately deprotonate. The high stability of the phenoxonium cation of alpha-tocopherol compared to the other tocopherols (and most other phenols) is a chemically important feature that may shed new light on understanding alpha-tocopherol's unique biological properties.     

Magneto-structural characterization of metallocene-bridged nitronyl nitroxide diradicals by X-Ray, magnetic measurements, solid-state NMR spectroscopy, and ab initio calculations

Crystallization of ferrocene and ruthenocene substituted in the 1- and 1'-positions by two nitronyl nitroxide radicals gave the new crystal phases beta-1 (besides the known phase alpha-1), alpha-2, and beta-2 whose structures were determined by X-ray analysis. In beta-1 the radical moieties adopt transoid positions, whereas two different cisoid conformations are adopted by alpha-2 and beta-2. These conformations result from inter- and intramolecular hydrogen bonds, respectively. All compounds experience antiferromagnetic interactions, and J/k(B) values up to -7 K have been found by fitting the experimental magnetic susceptibilities to a modified Bleaney-Bowers equation. The solid diradicals alpha-1, beta-1, alpha-2, and beta-2 as well as the ferrocene 3, which was substituted by a unique nitronyl nitroxide, were investigated by (13)C and (1)H NMR spectroscopy with magic angle spinning. The carbon signals cover a range of 2000 ppm, and are well resolved such that the structure could be confirmed. Conversion of the signal shifts into spin densities disclosed the mechanisms by which spin delocalization from the nitronyl nitroxide substituents to the metallocene core occurs. The spin density distribution in alpha-1, beta-1, and 3 was also predicted by DFT calculations. There is good agreement between the experimental and theoretical trends of the spin delocalization. The magnetic interactions were discussed in the light of intramolecular spin transfer and its dependence on geometric constraints, demonstrating that the 1,1'-metallocenylene bridge is not a robust magnetic coupler.     

Chromatographic separation of tocopherols.

alpha-, beta-, gamma-, and delta-Tocopherols were separated by reversed-phase high-performance thin-layer chromatography (C18RP-HPTLC), normal-phase high-performance liquid chromatography (NP-HPLC), reversed-phase high-performance liquid chromatography (C18RP-HPLC), and gas chromatography (GC). The selected topological indices based on connectivity (M, 1chi(v)), on distance matrix (W, (o)B, MTI) and on information theory (I(AC), I(AC)) were calculated for these tocopherols. The observed chromatographic separations of investigated tocopherols were compared. This comparison indicated that the C18RP-HPTLC, NP-HPLC, and GC are the best techniques for the separation of these tocopherols. Topological index (o)B was the most significant. We obtained definite dependence between the numerical values of topological index (o)B and the chromatographic separation of the investigated tocopherols.     

Simultaneous analysis of Vitamins A and E in infant milk-based formulae by normal-phase high-performance liquid chromatography-diode array detection using a short narrow-bore column

A rapid, simple and reproducible normal-phase (NP) high-performance liquid chromatography (HPLC)-diode array detection (DAD) method for simultaneous qualitative and quantitative determination of Vitamin A (retinol acetate and retinol palmitate) and Vitamin E (alpha-tocopherol acetate, alpha-, gamma- and delta-tocopherols) in milk-based infant formulae was developed and validated. The preparation sample was based on protein precipitation and vitamin extraction with ethanol, followed by re-extraction with hexane, while the chromatographic method was based on the use of a short narrow-bore column (50 mm x 2.1 mm; 3 microm particle size), which afforded less solvent consumption and higher mass sensitivity. The method showed acceptable values for precision, recovery and sensitivity, and proved very simple for routine analysis work.     

Terpsichorean movements of pentaammineruthenium on pyrimidine and isocytosine ligands

Pentaammineruthenium moves on ambidentate nitrogen heterocycles by both rotation and linkage isomerization, which may affect the biological activity of potential ruthenium metallopharmaceuticals. The rapid rotation rates of [(NH3)5RuIII] coordinated to the exocyclic nitrogens of isocytosine (ICyt) and 6-methylisocytosine (6MeICyt) have been determined by 1H NMR. Since these rotamers can be stabilized by hydrogen bonding between the coordinated ammines and the N1 and N3 endocyclic nitrogens, rotamerization is under pH control. Spectrophotometrically (UV-vis) measured pKa values for the two endocyclic sites for the ICyt complex are 2.78 and 9.98, and for 6MeICyt are 3.06 and 10.21, which are probably weighted averages for ionization from N3 and N1, respectively. Activation parameters for the rotamerizations were determined by variable-temperature NMR at pKa1 < pH < pKa2 for the complexes with (ICyt-kappa N2)-, (6MeICyt kappa N2)-, and 2AmPym kappa N2. For [(6MeICyt kappa N2)(-)-(NH3)5RuIII]2+, delta H* = 1.6 kcal/mol, delta S* = -37 cal/mol K, and Ea = 2.2 kcal/mol. Due to strong RuIII-N pi-bonding, the activation enthalpies are approximately 10 kcal lower than the expected values for the free ligands. Rotameric structure is correlated with pKa values, pH-dependent reduction potentials, and 1H NMR parameters. Linkage isomers of [(2AmPym)(NH3)5Ru]n+ are reported in which RuII is coordinated to the endocyclic nitrogen (N1) and RuIII to the exocyclic nitrogen (N2). The rate constant for the kappa N2-->kappa N1 isomerization as part of an ECE mechanism is 3.9 s-1 at pH 3. The pH dependence of the acid-catalyzed hydrolysis of [(2AmPym kappa N1)(NH3)5Ru]2+ is determined.     

Mixed liquids for single-drop microextraction of organochlorine pesticides in vegetables

A new approach for the extraction of nine kinds of organochlorine pesticides (OCPs) from vegetable samples coupling single-drop microextraction with gas chromatography-mass spectrometry was presented. Experimental parameters, such as organic solvent, exposure time, agitation and organic drop volume were controlled and optimized. An effective extraction was achieved by suspending a 1.00microL mixed drop of p-xylene and acetone (8:2, v/v) to the tip of a microsyringe immersed in a 2mL donor aqueous solution and stirred at 400rpm. The approach was applied to the determination of OCPs in vegetable samples with a linearity range of 0.05-20ng mL(-1) for alpha-, beta-, gamma-, delta-hexachlorobenzene (BHC) and dicofol, 0.5-20ng mL(-1) for dieldrin and 2,2-bis(4-chlorophenyl)-1,1-dichloroethane (DDD) or 0.5-50ng mL(-1) for 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (DDE) and 2-(2-chlorophenyl)-2 (4-chlorophenyl)-1,1,1-trichloroethane (p,p'-DDT). Correspondingly, the determination limit at an S/N of 3 ranged from 0.05ng mL(-1) for alpha-, beta-, gamma-, delta-BHC to 0.2ng mL(-1) for dicofol, dieldrin or p,p'-DDT. The relative recoveries were from 63.3 to 100%, with repeatability ranging from 8.74 to 18.9% (relative standard deviation, R.S.D.). The single-drop microextraction was proved to be a fast and simple approach for the pre-concentration of organochlorine pesticides in vegetable samples.