A fluorinated chitosan-based QuEChERS method for simultaneous determination of 20 organophosphorus pesticide residues in ginseng using GC-MS/MS.

In this study, a new fluorinated methacrylamide (MACF) was synthesized and evaluated as an adsorbent in the dispersive solid-phase extraction for the effective determination and extraction of 20 organophosphorus pesticides (OPPs) from ginseng samples using the QuEChERS (quick, easy, cheap, effective, rugged, safe) method coupled with GC-MS/MS. The properties of MACF were characterized using Fourier-transform infrared spectroscopy, elemental analysis, and high-resolution 19 F NMR. MACF, chitosan, primary and secondary amine, octadecylsilane, graphitized carbon black, Z-Sep, Z-Sep+ , and EMR-Lipid were compared in terms of extraction efficiency. The best results were obtained when MACF was used. Matrix-matched calibration was employed for quantification. All the OPPs exhibited good linearity (r2 > 0.9969) with the concentration at their respective concentration ranges. The limits of detection were 1.5-3.0 µg/kg, and the limits of quantification were 5.0-10.0 µg/kg. The trueness of the 20 pesticides at four spiked levels ranged from 86.1 to 111.1%, and the relative standard deviation was less than 11.3%. The modified QuEChERS method using MACF as the adsorbent was sensitive, reliable, and cost-effective and could be used for the determination of 20 OPP residues in ginseng.

Triphenylphosphonium functionalized Ficus religiosa L. extract loaded nanoparticles improve the mitochondrial function in oxidative stress induced diabetes.

The present study was aimed to enhance the mitochondrial function in oxidative stress-induced diabetes. To achieve this, Ficus religiosa L. extract loaded solid lipid nanoparticles (ETNPs) were prepared and functionalized by using triphenylphosphonium. Developed nanoparticles demonstrated desired quality attributes with sustained release for up to 24 h and excellent storage stability for up to 180 days at 40 ± 2°C and 75 ± 5% relative humidity. In vitro cytotoxicity assessment showed no toxicity of ETNPs. Interestingly, oral administration of ETNPs to diabetic rats demonstrated improved mitochondrial function by normalizing the mitochondrial morphology, intracellular calcium ion concentration, complexes I, II, IV, and V activity, mitochondrial membrane potential, and antioxidant levels. Further, reduction in apoptotic markers viz. cytochrome-C, caspase-3, and caspase-9 was observed following the ETNP treatment. Moreover, significant reduction in blood glucose and glycosylated hemoglobin while significant improvement in plasma insulin was observed as compared to the diabetic group following the treatment with developed formulation. Furthermore, histopathology studies confirmed the safety of the developed formulation and thus, data in hand collectively suggest that proposed strategy can be effectively used to improve the mitochondrial function in oxidative stress-induced diabetes along with better control over blood glucose and glycosylated hemoglobin.

Screening of chiral shift reagents suitable to generically separate the enantiomers of V-agents by 31P-NMR spectroscopy.

Fourteen amino acids protected at the N-terminal and at their side chains were screened for resolving the enantiomers of V-agents by NMR. While none of the shift reagents tested showed really effective separation in proton NMR, two of them (BOC-Gln(Xan)-OH, 16, and Z-Arg(Z)2-OH), 21, with 16 superior to 21) were found suitable to separate the enantiomers of all V-agent homologues involved in the test by 31P-NMR. Molar ratios investigated were 1:0.5, 1:1, 1:1.5, 1:2, and 1:3 with the V-agent set to 1 throughout the experiments. All these ratios were more or less effective, but 1:3 was found to separate the V-agents the most reliable way. It is postulated that three chiral solvating molecules are then coordinated around the organophosphate: ion pair formation with the amino nitrogen of the V agent side chain, hydrogen bonding provided by the PO unit, and extension of coordination at the phosphorus atom itself. After chiral separation of VX by semi-preparative LC-MS the enantiomers were examined with both configurations of 16 releasing four different 31P NMR peaks which correspond to four different complexes: R-S3, R-R3, S-R3, and S-S3. Comparing these results with literature data it is assumed that (+)-VX corresponds to the RP configuration and (-)-VX to the SP-configuration.

Dispersive Solid-Phase Extraction Using Microporous Sorbent UiO-66 Coupled to Gas Chromatography-Tandem Mass Spectrometry: A QuEChERS-Type Method for the Determination of Organophosphorus Pesticide Residues in Edible Vegetable Oils without Matrix Interference.

A QuEChERS-type method without matrix interference was designed and developed to determine organophosphorus pesticide residues in edible vegetable oils, based on dispersive solid-phase extraction with cleanup using UiO-66 as sorbent. Microporous UiO-66 directly and selectively adsorbed organophosphorus pesticides and excluded interfering compounds. Clean analytes were obtained by elution and analyzed using gas chromatography-tandem mass spectrometry. The dispersive solid-phase extraction conditions (amount of adsorbent, extraction time, desorption solvent volume, and elution time) were optimized. The limits of detection of the pesticides in vegetable oils were 0.16-1.56 ng/g. Under optimized conditions, the average pesticide recoveries were 81.1-113.5%. The intraday and interday relative standard deviations for analyte recovery were <8.2 and <13.9%, respectively. Thus, the method is reliable and could detect organophosphorus pesticide residues in edible vegetable oils. Furthermore, UiO-66 can be easily recycled and reused at least 10 times, reducing the cost of analysis.

Sensitive Detection of Organophosphorus Pesticides in Medicinal Plants Using Ultrasound-Assisted Dispersive Liquid-Liquid Microextraction Combined with Sweeping Micellar Electrokinetic Chromatography.

A simple, rapid, and sensitive method using ultrasound-assisted dispersive liquid-liquid microextraction (UA-DLLME) combined with sweeping micellar electrokinetic chromatography (sweeping-MEKC) has been developed for the determination of nine organophosphorus pesticides (chlorfenvinphos, parathion, quinalphos, fenitrothion, azinphos-ethyl, parathion-methyl, fensulfothion, methidathion, and paraoxon). The important parameters that affect the UA-DLLME and sweeping efficiency were investigated. Under the optimized conditions, the proposed method provided 779.0-6203.5-fold enrichment of the nine pesticides compared to the normal MEKC method. The limits of detection ranged from 0.002 to 0.008 mg kg(-1). The relative standard deviations of the peak area ranged from 1.2 to 6.5%, indicating the good repeatability of the method. Finally, the developed UA-DLLME-sweeping-MEKC method has been successfully applied to the analysis of the investigated pesticides in several medicinal plants, including Lycium chinense, Dioscorea opposite, Codonopsis pilosula, and Panax ginseng, indicating that this method is suitable for the determination of trace pesticide residues in real samples with complex matrices.

An improved dispersive solid-phase extraction clean-up method for the gas chromatography-negative chemical ionisation tandem mass spectrometric determination of multiclass pesticide residues in edible oils.

An improved sample preparation using dispersive solid-phase extraction clean-up was proposed for the trace level determination of 35 multiclass pesticide residues (organochlorine, organophosphorus and synthetic pyrethroids) in edible oils. Quantification of the analytes was carried out by gas chromatography-mass spectrometry in negative chemical ionisation mode (GC-NCI-MS/MS). The limit of detection and limit of quantification of residues were in the range of 0.01-1ng/g and 0.05-2ng/g, respectively. The analytes showed recoveries between 62% and 110%, and the matrix effect was observed to be less than 25% for most of the pesticides. Crude edible oil samples showed endosulfan isomers, p,p'-DDD, α-cypermethrin, chlorpyrifos, and diazinon residues in the range of 0.56-2.14ng/g. However, no pesticide residues in the detection range of the method were observed in refined oils.

Highly sensitive and selective organophosphate screening in twelve commodities of fruits, vegetables and herbal medicines by dispersive liquid-liquid microextraction.

The article describes a simple sample pretreatment procedure for the analysis of ten organophosphorus pesticides using dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC-MS) in three distinctively different types of matrices: fresh fruits, fresh vegetables and dried herbs. The method was carefully developed, focusing on the chemistry of various dispersive solvents, to achieve simultaneous, comprehensive extraction and preconcentration in a great span of selected matrices. According to matrix-matched validation study, the set of optimized DLLME conditions has been proven robust to determine target OPPs within a wide linear range from 0.1 to 1000 µg L(-1). With limited usage of organic extractants, remarkable enrichment factors up to 100-fold were obtained, enabling ultra-trace pesticide quantification down to sub-ppt levels at 0.12-4.92 ng kg(-1). Practical application of the method was illustrated by quantitative recovery (70-119%) and good precision (2.6-10% R.S.D.) in a representative range of three fruits and four vegetable commodities featured by the CODEX Alimentarius classification as well as their unique matrix compositions. A careful selection of dried herbs was further classified based on their morphological structures to validate analytical ruggedness of the method. Compared with existing methods for food analysis vis-à-vis OPPs, the present method is superior in terms of high sample throughput, minimal solvent consumption, and small sample size requirement. An additional, significant aspect of this universal DLLME method is that it models sample pretreatment methods with wide coverage of analytical matrices that are more effective, more comprehensive, and more flexible than those currently being used.

Simultaneous determination of organophosphorus, organochlorine, pyrethriod and carbamate pesticides in Radix astragali by microwave-assisted extraction/dispersive-solid phase extraction coupled with GC-MS.

A simple, rapid and reliable method was proposed for the simultaneous determination of 27 pesticides (organophosphorus, organochlorine, pyrethroid and carbamate pesticides) in Radix astragali. The pesticides were extracted by acetonitrile and the experimental variables, such as temperature, extraction time and volume of acetonitrile, were optimized through orthogonal array experimental design. Cleanup of extracts was performed with dispersive-solid phase extraction using primary secondary amine (PSA) as the sorbent. The determination of pesticides in the final extracts was carried out by gas chromatography-mass spectrometry in selected ion monitoring mode (GC-MS, SIM). The linearity of the calibration curves is good in matrix-matched standard, and yields the coefficients of determination (R(2))≥0.99 for approximately 96% of the target analytes. Under optimized conditions, the average recoveries (six replicates) for most pesticides (spiked at 0.02, 0.1 and 0.2 mg kg(-1)) range from 70% to 120%, and RSDs are less than 17.2%.

Microwave accelerated selective Soxhlet extraction for the determination of organophosphorus and carbamate pesticides in ginseng with gas chromatography/mass spectrometry.

Microwave accelerated selective Soxhlet extraction (MA-SSE), a novel selective extraction technique, was investigated in this study. A Soxhlet extraction system containing a glass filter was designed as an extractor. During the procedure of MA-SSE, both the target analytes and the interfering components were extracted from the sample into the extraction solvent enhanced by microwave irradiation. After the solvent flowed though the sorbent, the interfering components were adsorbed by the sorbent, and the target analytes remaining in the solvent were collected in the extraction bottle. No cleanup or filtration was required after extraction. The efficiency of the MA-SSE approach was demonstrated in the determination of organophosphorus and carbamate pesticide residues in ginseng by gas chromatography/mass spectrometry (GC/MS). Under the optimized conditions, low limits of detection (0.050-0.50 µg/kg) were obtained. The recoveries were in the range of 72.0-110.1% with relative standard deviations less than 7.1%. Because of the effect of microwave irradiation, MA-SSE showed significant advantage compared with other extraction techniques. The sorbent used in this study showed good cleanup ability. The mechanism of MA-SSE was demonstrated to be based on the rupture of the cell walls according to the structural changes of ginseng samples. On the basis of the results, MA-SSE as a simple and effective sample preparation technique for the analysis of pesticide residues in complex matrixes shows great promise.

Evaluation of microwave-assisted extraction for the analysis of organophosphorus and pyrethroid pesticides in green onions.

The aim of this study was to evaluate the efficiency of microwave-assisted extraction (MAE) for the analysis of organophosphorus (OP) and pyrethroid (PYR) pesticides in green onions by GC/MS. We optimized MAE extraction solvent, temperature, and time by using a certified reference material. As a result, the concentrations of two OP and two PYR target pesticides obtained by MAE with acetonitrile at 110 degrees C for 10 min were in good agreement with certified concentrations and comparable to the results by homogenization used as reference extraction technique. When the recovery test, performed by spiking the target pesticides into blank samples (5.0 g), was carried out with our optimized MAE conditions, mean recoveries of 16 OP and 10 PYR pesticides were 72-108% for a 1.0 pg spiking level and 70-119% for a 0.2 microg level. These results were acceptable according to the validation guideline for testing method of agricultural chemicals in food by Ministry of Health, Labour, and Welfare in Japan. The results suggested that MAE can be used for the analysis of OP and PYR pesticides in green onions.

One-step extraction for gas chromatography with flame photometric detection of 18 organophosphorus pesticides in Chinese medicine health wines.

An easy, rapid and selective gas chromatography with flame photometric detection (GC-FPD) method was established for simultaneously determining 18 organophosphorus pesticides (OPPs) in 80 Chinese medicine (CM) health wines. This method was based on a simple one-step extraction procedure using a little solvent without any further cleanup steps. The optimized extraction solvent for the pesticides is acetone:dichloromethane (1:1, V/V) with extraction recovery of 79.0-109.1% and relative standard deviation (RSD) of 0.36-12.68%, respectively. The limits of detection (LODs) of the established GC-FPD method for all investigated pesticides ranged from 1 to 15ngmL(-1) and limits of quantification (LOQs) from 4 to 50ngmL(-1). Out of all 80 CM health wines, 18 OPPs were found in 8 samples at low concentrations of 8.2-37.9ngmL(-1). These pesticides were successfully confirmed by GC-MS. This is the first report of determining OPPs in CM health wines, providing references for monitoring the quality of CM health wine in routine analysis.

[Studies on the chemical constituents of Rubus parvifolius].

OBJECTIVE: To study the chemical constituents of Rubus parvifoliu. METHODS: The constituents were isolated by column chromatography and their structures were elucidated through spectroscopic analysis such as 1H-NMR, 13C-NMR, FT-IR, et al. RESULTS: Seven compounds were isolated from the roots of Rubus parvifolius L., they were identified as p-sitosterol (I), lauric acid (II), O-nitrophenol (III), beta-daucosterol (IV), euscaphic acid (V), camelliagenin A (VI) and(+) -catech in (VII). CONCLUSION: Compounds III and VII are isolated from the plant for the first time.

Organohalogen and organophosphorous pesticide method for ginseng root--a comparison of gas chromatography-single quadrupole mass spectrometry with high resolution time-of-flight mass spectrometry.

A method has been developed for the analysis of 170 organohalogen and organophosphorous pesticides, isomers, and metabolites in dried ground ginseng root. Pesticides were extracted with ethyl acetate and purified with gel permeation chromatography (GPC) and primary/secondary amine modified silica (PSA)/graphitized carbon black (GCB) combination SPE column. Each purified pesticide extract was determined by both gas chromatography single quadrupole mass spectrometry using selected ion monitoring (GC-qMS-SIM) and by gas chromatography high resolution time-of-flight mass spectrometry (GC-HR-TOFMS). The geometric mean LOQs using the qMS and TOFMS were 4 and 3 ng/g ginseng, respectively. Mean recoveries from ginseng were 83, 79, and 75% with standard deviations of 4, 5, and 3%, respectively, for 25, 100, and 500 ng/g using GC-qMS-SIM. Mean recoveries using GC-HR-TOFMS were 93, 85, and 81% with mean standard deviations of 7, 7, and 8% for 25, 100, and 500 ng/g, respectively. Seven dried ginseng root products were found to contain combinations of the following pesticides: dacthal, diazinon, DDT, hexachlorobenzene, iprodione, lindane, procymidone, and quintozene (1-460 ng/g). No significant differences were found in the concentrations measured for these pesticides on commercial ginsengs using either of the two GC/MS techniques.

Application of statistical design of experiment with desirability function for the removal of organophosphorus pesticide from aqueous solution by low-cost material.

This paper deals with the multiple response optimization for the removal of organophosphorus pesticide quinalphos [QP: O,O-diethyl O-2-quinoxalinyl phosphorothioate] from the aqueous solution onto low-cost material and tried to overcome the drawbacks of univariate optimization. Used tea leaves were used as low-cost adsorbent and batch equilibration method was followed for this study. A Box-Behnken design was used to develop response model and desirability function was then used for simultaneous optimization of all affecting parameters in order to achieve the highest removal% of quinalphos. The optimum conditions of factors predicted for quinalphos removal% were found to be: pH 8.83, concentration 7 mg L(-1) and dose 0.40 g. Under these conditions, maximum removal% of quinalphos was obtained 96.31%. Considering the above optimum conditions, the adsorption isotherms were developed and provided adsorption capacity of 196.07 microg g(-1) by using Langmuir equation, indicating that used tea leaves may be applied as a low-cost material for pesticides removal from aqueous matrices.

Development of dispersive liquid-liquid microextraction method for the analysis of organophosphorus pesticides in tea.

In this article, a new method for the determination of organophosphorus pesticides (OPPs) in tea was developed by using dispersive liquid-liquid microextraction (DLLME) and gas chromatography-flame photometric detection (GC-FPD). A mixture of acetonitrile and n-hexane was used as an extraction solvent for the extraction of OPPs from tea samples. When the extraction process was finished, the mixture of solvents was rapidly dispersed in water; target analyte was extracted to a small volume of n-hexane, using DLLME. Recovery tests were performed for concentration 5.0 microg/kg. The recovery for each target analyte was in the range between 83.3 and 117.4%. The repeatability of the proposed method, expressed as relative standard deviation, varied between 3 and 7.8% (n=3). The detection limit of the method for tea was found ranging from 0.030 to 1 microg/kg for all the target pesticides. Compared with the conventional sample preparation method, the proposed method has the advantage of being quick and easy to operate, and has high-enrichment factors and low consumption of organic solvent.

Removal of two organophosphate pesticides by a bacterial consortium immobilized in alginate or tezontle.

In order to remove methyl-parathion (MP) and tetrachlorvinphos (TCF), a bacterial consortium was immobilized with two supports consisting of alginate beads or stones of tezontle colonized by biofilm. Removal kinetics were recorded for suspended and immobilized consortium using a mineral salt medium supplemented with MP and TCF at 25mg/L and with 0.1% (w/v) glucose as a co-substrate. The viability of the consortium cultivated in suspension was maintained for 6 days, whereas the viability of the consortium immobilized in alginate and tezontle supports was maintained for up to 11 and 13 days, respectively. Growth was enhanced when using glucose as a co-substrate. The percentage of MP removed was significantly higher (alpha=0.05) when consortium was immobilized in alginate beads and biofilm on tezontle as compared to suspension culture.

Aeruginascin, a trimethylammonium analogue of psilocybin from the hallucinogenic mushroom Inocybe aeruginascens.

The hallucinogenic mushroom Inocybe aeruginascens contains several typical Psilocybe alkaloids including psilocybin. We have now elucidated the structure of a further indole derivative named aeruginascin as the quaternary ammonium compound N, N, N-trimethyl-4-phosphoryloxytryptamine. Aeruginascin is closely related to the frog skin toxin bufotenidine (5-HTQ), a potent 5-HT3 receptor agonist, and has been found exclusively in Inocybe aeruginascens so far.

Analysis of phosphorus herbicides by ion-pairing reversed-phase liquid chromatography coupled to inductively coupled plasma mass spectrometry with octapole reaction cell.

A reversed phase ion-pairing high performance liquid chromatographic (RPIP-HPLC) method is developed for the separation of two phosphorus herbicides, Glufosinate and Glyphosate as well as Aminomethylphosphonic acid (AMPA), the major metabolite of Glyphosate. Tetrabutylammonium hydroxide is used as the ion-pairing reagent in conjunction with an ammonium acetate/acetic acid buffering system at pH 4.7. An inductively coupled plasma mass spectrometer (ICP-MS) is coupled to the chromatographic system to detect the herbicides at m/z = 31P. Historically, phosphorus has been recognized as one of the elements difficult to analyze in argon plasma. This is due to its relatively high ionization potential (10.5 eV) as well as the inherent presence of the polyatomic interferences 14N16O1H+ and 15N16O+ overlapping its only isotope at m/z = 31. An octapole reaction cell is utilized to minimize the isobaric polyatomic interferences and to obtain the highest signal-to-background ratio. Detection limits were found to be in the low ppt range (25-32 ng/l). The developed method is successfully applied to the analysis of water samples collected from the Ohio River and spiked with a standard compounds at a level of 20 microg/l.

Oligonucleotide N-alkylphosphoramidates: synthesis and binding to polynucleotides.

A few different methods for the preparation of oligonucleotide N-alkylphosphoramidates were compared directly. One of these, involving the use of protected nucleoside phosphites as building blocks, provided the requisite N-alkylphosphoramidates via oxidation of the intermediate dinucleoside methyl phosphites with iodine in the presence of the appropriate alkylamine. This method was found to have several attractive features, including the use of building blocks identical with those employed for the synthesis of DNA and compatibility with procedures and instruments employed for the stepwise synthesis of oligonucleotides by solution and solid-phase methods. This procedure was used to make several di-, tri-, and tetranucleotide N-alkylphosphoramidates derived from deoxyadenosine and thymidine; alkyl substituents included N,N-dimethyl, N-butyl, N-octyl, N-dodecyl, and N-(5-aminopentyl). The aminoalkyl derivative of d(TpT) (24) was used to demonstrate the feasibility of introducing an intercalative agent to the alkylphosphoramidate moiety of such derivatives. The oligonucleotide N-alkylphosphoramidates were separated into their component diastereomers and characterized structurally by a number of techniques including circular dichroism, high-field 1H NMR spectroscopy, FAB mass spectrometry, and enzymatic digestion to authentic nucleosides and nucleotides. Physicochemical characterization of several di- and trinucleotide alkyl-phosphoramidates revealed that the adenine nucleotide analogues formed stable complexes with poly-(thymidylic acid). The stabilities of these complexes were found to increase with increasing chain length of the N-alkylphosphoramidate substituents. The finding that N-alkylphosphoramidate substituents can enhance the binding of certain oligonucleotides to their complementary polynucleotides suggests the existence of a novel source of polynucleotide affinity.

Selective retention of organic phosphate esters and phosphonates on aluminium oxide.

Compounds containing the -PO3H2 function, such as monoesters of phosphoric acid and phosphonic acids, specifically bind to aluminium oxide in aqueous solution under experimental conditions where non-phosphorylated compounds are completely desorbed. The bound organic phosphate can be specifically displaced by aqueous solution of inorganic phosphates thus allowing their separation or detection by a technique similar to that of affinity chromatography. The consequences of this finding for phosphate compound biochemistry are discussed.