Determination of 2-alkylcyclobutanones by combining precolumn derivatization with 1-naphthalenyl hydrazine and ultra-performance liquid chromatography with fluorescence detection.

2-Alkylcyclobutanones (2-ACBs) are uniquely formed when triglycerides-containing food products are exposed to ionizing radiation. Thus, 2-ACBs have been used as marker molecules to identify irradiated food. Most methods to determine 2-ACBs involve mass spectrometric detection after chromatographic separation. The spectrofluorometer is rarely used to determine 2-ACBs because these molecules do not fluoresce. In this study, we developed an ultra-performance liquid chromatography (UPLC) method to determine 2-ACBs. 2-ACBs were converted into fluorophores after reacting with 1-naphthalenyl hydrazine to facilitate their sensitive and selective detection using a fluorescence detector (FLD). Analysis of 2-ACBs using our developed UPLC-FLD method allows sensitive determination of 2-ACBs at a detection limit of 2 ng 2-ACBs per g of fat (30 pg/injection), which is significantly lower than that of existing analytical methods. After validation for trueness and precision, the method was applied to γ-irradiated chicken samples to determine their 2-ACB content. Comparative studies employing liquid chromatography-tandem mass spectrometric method revealed no systematic difference between the two methods, thereby demonstrating that the proposed UPLC-FLD method can be suitably used to determine 2-ACBs in irradiated foodstuffs. Graphical Abstract Determination of radiation-induced food-borne 2-dodecylcyclobutanone and 2-tetradecylcyclobutanone by combining 1-naphthalenyl hydrazine derivatization and ultra-performance liquid chromatography with fluorescence detection.

Alterations of endogenous metabolites in urine of rats exposed to decabromodiphenyl ether using metabonomic approaches.

There is large usage of polybrominated diphenyl ethers (PBDEs) especially for decabromodiphenyl ether (BDE-209, Deca-BDE) in controlling the risks of fire. The toxicological effects of PBDEs are worth being concerned about. Female SD rats were daily gavaged with BDE-209 ether at the dose of 100 mg/kg for 20 days. Histological observation was performed for the screening of the target organs for BDE-209 exposure. The distribution and metabolism of PBDEs in the exposed main organs were evidenced by HRGC-HRMS. Alterations of the endogenous metabolite concentrations in urine were investigated using metabonomic approaches based on (1)H NMR spectrum. Histopathological changes including serious edema in kidney, hepatocellular spotty necrosis and perivasculitis in liver indicated that BDE-209 caused potential influences on endogenous metabolism in the exposed liver and the kidney. BDE-209 was found to be highly accumulated in lipid, ovary, kidney and liver after 20 days' exposure. Occurrence of other lower brominated PBDEs in the rats demonstrated that reductive debromination process happened in vivo. Hydroxylated and methoxylated-BDEs, as metabolism products, were also detected in the rat tissues. A total of 12 different endogenous metabolites showed obvious alterations in urine from the exposed rats, indicating the disturbance of the corresponding internal biochemical processes induced by BDE-209 exposure. These findings in vivo suggested the potential health risk might be of concern due to the toxicological effects of BDE-209 as a ubiquitous compound in the environment.

Photo-controllable molecular hydrogels for drug delivery.

A photocleavable nitrobenzyl ester group was introduced in the backbone of self-assembling peptides using standard Fmoc-peptide systhesis. The nitrobenzyl ester group could be used as a cleavable linker to connect a gelator and a hydrophilic group. Photo-irradiation would lead to the generation of the molecular gelator and the formation of a molecular hydrogel. The nitrobenzyl ester group could also be used as one part of hydrophobic capping group for short peptides to produce molecular hydrogelators. In this case, photo-irradiation would lead to the release of hydrophobic anti-imflammatory drug of Naproxen and the gel-sol phase transition. The application of photocleavable group in molecular self-assembly would lead to photo-controllable self-assembly materials for controlled drug release, tissue engineering, and regenerative medicine.

Liquid chromatography-electrospray ionization tandem mass spectrometric analysis of 2-alkylcyclobutanones in irradiated chicken by precolumn derivatization with hydroxylamine.

Food irradiation is a common preservation method that is used in many countries. The ability to identify irradiated food is important for assuring compliance with regulatory policies, such as food labeling requirements, and for informed consumer choice. There is thus a significant demand for analytical methods of high sensitivity and selectivity to identify irradiated food, especially for foods subjected to low-dose irradiation and for processed or composite foods that contain small quantities of irradiated ingredients. 2-Alkylcyclobutanones (2-ACBs) are uniquely formed during food irradiation and have been adopted by the European Committee for Standardization as signature biomarkers for the identification of irradiated foods. We now report the development of a novel assay for quantification of 2-ACBs in γ-irradiated food by liquid extraction of fat content followed by precolumn derivatization and liquid chromatography-tandem mass spectrometric (LC-MS/MS) detection. Precolumn derivatization with hydroxylamine introduced a polar functional group into the otherwise nonpolar 2-ACBs, which greatly enhanced ESI-MS response. The method was validated for extraction efficiency, precision, accuracy, and detection limit. In comparison with the current GC-MS based European official method (EN1785:2003) for 2-ACBs determination, our new LC-MS/MS method offers a more efficient sample processing protocol with reduced solvent consumption. More importantly, the combination of chemical derivatization and LC-MS/MS detection significantly enhanced the analytical sensitivity of the method, which allows confident identification of food irradiated with as little as 10 Gy. To the best of our knowledge, this is the first report of 2-ACB determination by LC-MS/MS and the first analytical method allowing confident identification of irradiated food at dosage of down to 10 Gy.

Glutathione-triggered formation of molecular hydrogels for 3D cell culture.

The development of three dimensional (3D) scaffolds that are suitable for cell encapsulation and proliferation is highly important for tissue engineering and regenerative medicine. We reported in this paper on several molecular hydrogels formed through glutathione (GSH) reduction, whose mechanical property and zeta potential could be regulated by concentration and structure of gelators in resulting gels, respectively. The hydrogels were characterized by several techniques including rheology, TEM and fluorescence. We found that, in our system, the mechanical property of hydrogels but not the zeta potential of self-assembled structures had big influences on mouse fibroblast 3T3 cells spreading and proliferation. Hydrogels with storage modulus (G') of hundreds of pascals (Pa) were suitable for 3T3 cells spreading and proliferation. We believed that hydrogels reported in this study had big potential for applications in different fields, such as 3D cell culture and tissue engineering.

A molecular hydrogel of a camptothecin derivative.

We report on the first example of self-assembled nanotubes of a camptothecin derivative in a molecular hydrogel.

[Rapid determination of 40 pesticide residues in fruits using gas chromatography-mass spectrometry coupled with analyte protectants to compensate for matrix effects].

A gas chromatography-mass spectrometry (GC-MS) method was developed for the determination of 40 pesticides in fruits. The effects of adding analyte protectants were evaluated for compensating matrix effects and the impacts on the quantitative results. A new combination of analyte protectants - Polyethylene Glycol 400 (PEG 400) and olive oil combination, which can be dissolved in acetone, was used for the quantitative analysis. The pesticides were extracted from fruit samples with acetonitrile and the extracts were cleaned up using micro-solid phase extraction. A GC-MS method in selective ion monitoring (SIM) mode coupled with large volume injection was finally developed. Using the newly developed analyte protectant combination of PEG 400 and olive oil, a good linearity was obtained in the range of 1 - 200 microg/L with coefficients better than 0.99, and the detection limits were between 0.1 - 3.0 microg/L. The mean recoveries of the pesticides were 75% - 119% with the relative standard deviation values less than 16.6% except for dimethoate. The performance of the analyte protectants was compared with matrix-matched standards calibration curves in terms of quantitative accuracy. The results showed that the method of adding analyte protectants can replace the matrix-matched standard calibration, and can also reduce the sample pretreatment. When the devel- oped method was used for the analysis of apple, peache, orange, banana, grape and other fruit samples, a good matrix compensation effect was achieved, and thus effectively reduced the bad effects of the water-soluble agents to the gas chromatographic column.

[Determination of dioxin like polychlorinated biphenyl residues in milk using isotope dilution gas chromatography-ion trap tandem mass spectrometry].

A method for the determination of dioxin like polychlorinated biphenyl (DL-PCB) residues in milk has been developed using high resolution gas chromatography coupled with ion trap mass spectrometry (GC-MS/MS-IT). Analytical procedure consisted of accelerate solvent extraction (ASE), lipid removal with acidic silica and clean-up using anthropogenic isolation column packed with multilayer silica. The analytes were separated on a DB-5 capillary column, detected with multiple reaction monitor mode (MRM) and quantified using internal standard calibration curve of isotope dilution technique. The correlation coefficients of calibration standard solution were above 0. 999 9 for all the DL-PCBs. The recoveries and relative standard deviations of labeled compound solution were in the oranges from 39% to 129% and from 5% to 22%, respectively. The detection limits in the range from 3 to 11 pg/g (fat) were established for the 12 DL-PCBs.

E-waste recycling induced polybrominated diphenyl ethers, polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and dibenzo-furans pollution in the ambient environment.

Polybrominated diphenyl ethers (PBDEs) have been determined in 30 samples, including soil, biota and plant collected from an electronic waste recycling site and its vicinage towns. Polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins and dibenzo-furans (PCDD/Fs) were also simultaneously analyzed in 20 samples. PBDEs were detected in all soil samples and the highest was up to 789 ng/g dry weight (dw). The toxic equivalency (TEQ) of PCBs and PCDD/Fs detected in E-waste recycling site is significant higher than those in the vicinage samples. Biota and plant were also contaminated with high level of PBDEs, PCBs and PCDD/Fs in this area. High levels of the three kinds of organic compounds in the environmental samples showed that the E-waste recycling have induced serious environmental problems.

Polybrominated diphenyl ethers and organochlorine pesticides in sewage sludge of wastewater treatment plants in China.

The concentrations of chemicals in sewage sludge from wastewater treatment plants (WWTPs) may relate to their levels of use and environmental pollution in the region. In this work, sludge samples were collected from 31 WWTPs in 26 cities in China and analyzed for polybrominated diphenyl ethers (PBDEs) and organochlorine pesticides (OCPs). The concentrations of SigmaPBDE (sum of congeners 17, 28, 47, 66, 71, 85, 99, 100, 138, 153, 154, and 183) ranged from 6.2 to 57ng/g (dw). The concentration of decabromodiphenyl ether (BDE-209) ranged from below limit of detection (LOD) to 1109ng/g (dw) (with a median of 27ng/g (dw)), and averaged 55% (median 69%) of the total PBDEs. These levels are about 10-100 times lower than those found in Europe and North America. PBDE levels in sludge were not found to depend on the location and capacity of the WWTPs. Concentrations of hexachlorocyclohexanes (HCHs), dichlorodiphenyltrichloroethane (DDTs) and its major degradation products, and hexachlorobenzene (HCB) ranged from below detection limit to 167ng/g (dw), 11 to 1065ng/g (dw), and 7.5 to 319ng/g (dw), respectively. The major DDT degradation products were p,p'-DDE and p,p'-DDD. The major hexachlorocyclohexane (HCH) isomer in sludge is beta-HCH, reflecting its higher affinity to solids and resistance to degradation than other isomers.

Occurrence of polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls pollution in sediments from the Haihe River and Dagu Drainage River in Tianjin City, China.

The pollution status of polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/Fs) and polychlorinated biphenyls (PCBs) in the sediments of Haihe River, which is the most polluted among the seven largest basins in China, Dagu Drainage River flowing through a chemical industry zone, and two other rivers flowing into Bohai Sea in Tianjin City, China were investigated. The concentrations of PCDD/Fs and PCBs in the sediments from the mainstream of Haihe River were 1.3-26pgI-TEQg(-1) dry weight (dw) and 0.07-0.54pgTEQg(-1)dw, respectively. Heavy PCDD/Fs and PCBs pollution, with 1264pgI-TEQg(-1)dw and 21pgTEQg(-1)dw, was found in sediment from Dagu Drainage River. The congener profiles of PCDD/Fs indicated that the principal contamination source was the production of pentachlorophenol (PCP) or PCP-Na in this area. The correlation between PCDD/Fs or PCBs and total organic matter (TOM) showed that PCDD/Fs or PCBs were independent on TOM.

Quantitative structure-activity relationship for prediction of the toxicity of polybrominated diphenyl ether (PBDE) congeners.

Levels of Polybrominated diphenyl ether (PBDEs) are increasing in the environment due to their use as flame retardants. The similarities of structure to polychlorinated biphenyl (PCB) congeners suggest that they may share similar toxicological properties, such as hepatic enzyme induction. In this work, quantitative structure-activity relationship (QSAR) models were constructed based on 406 descriptors for the logarithm of toxicology index (aryl hydrocarbon receptor relative binding affinities, AhR, I) of 18 PBDE congeners. The method used for building model is the Heuristic method, which is included in comprehensive descriptors for structural and statistical analysis (CODESSA) software. The best regression model involved four descriptors, which were related to the conformational changes, atomic reactivity, molecular electrostatic field, and non-uniformity of mass distribution in a molecule of PBDEs, etc. The high square of the correlation coefficient R(2)(0.903) showed the model was satisfactory.

Development of quantitative structure gas chromatographic relative retention time models on seven stationary phases for 209 polybrominated diphenyl ether congeners.

Quantitative structure retention relationships (QSRRs) were developed to predict the gas chromatographic (GC) relative retention times (RRTs) for 209 polybrominated diphenyl ether (PBDE) congeners using the heuristic method included in the computer software Comprehensive Descriptors for Structural and Statistical Analysis (CODESSA). A total of 445 constitutional, topological, geometrical, electrostatic, and semi-empirical quantum chemical descriptors were derived for all PBDEs. Using experimental RRT data for 126 PBDE congeners from the literature, predictive regression models were built for seven individual GC capillary columns differing in stationary phases. Each model includes four descriptors which included Wiener index, Randic index, polarity parameter, etc., selected by CODESSA. High predictability was obtained. High multiple correlation coefficients R(2) indicated that >98.5% (except for stationary phase CP-Sil 19) of the total variation in the predicted RRTs is explained by the fitted models. The models were subsequently used to predict the RRTs of the remaining 83 PBDE congeners on seven different stationary phases. The statistical results show that, compared with others, DB-XLB column not only produces the least number of peak overlaps but also results in shorter retention times.

AhR-active compounds in sediments of the Haihe and Dagu Rivers, China.

Total concentrations of compounds that can cause activation of the aryl hydrocarbon receptor (AhR) in extracts of river sediments from various locations in the Haihe and Dagu Rivers, Tianjin, China were determined by use of the in vitro H4IIE-luc cell line. AhR-active compounds were isolated from sediments by Soxhlet extraction, and the crude extracts were fractionated using a Florisil column into three Fractions. The response of samples was compared to the TCDD standard and expressed as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TCDD-EQs). Significant dioxin-like activity was observed in each sample. The TCDD-EQs in crude extracts of sediments (SCEs), as determined with the bioassay were 2-4 times greater than the sum of the TCDD-EQs of the eluent from fractions separated with a Florisil column. The results also showed that Fractions 2 and 3 contained most of the AhR-mediated activity. The results obtained by using the bioassay were compared with those of previous measurements of the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) and the structurally-similar AhR-active polychlorinated biphenyls (PCBs). It was determined that sediments from the Dagu River contained greater concentrations of TCDD-EQ than did sediments from the Haihe River except at Jingangqiao (location R3), which is associated with industrial activities in the adjacent densely populated area. The concentrations of TCDD-EQs, based on the EC20 and the relative potency ranges, of SCEs ranged from 330 to 930 and 1200 to 13,900 pg TCDD-EQ g(-1) dry wt in Haihe and Dagu Rivers, respectively.

Method development for the analysis of polybrominated diphenyl ethers, polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and dibenzo-furans in single extract of sediment samples.

A comprehensive method was developed for quantitative analysis of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins and dibenzo-furans (PCDD/Fs) in one single extract of environmental samples. The sample preparation procedure included two fractionation steps using silver nitrate silica chromatography to separate PBDEs from PCBs and PCDD/Fs and florisil column to separate PCBs from PCDD/Fs. Acidic silica, acidic alumina and gel permeation chromatography (GPC, for PCBs) or activated carbon column (for PCDD/Fs) were used for further clean-up. The sample extracts were analyzed by using high-resolution gas chromatography/high-resolution mass spectrometry. The entire method was validated from the analysis of mixed standards of PBDEs, PCBs and PCDD/Fs (n=3); the analysis of certified reference biota (WMF-01). The method was applied for the analysis of 10 sediment samples collected from Haihe River and Dagu Drainage River in Tianjin City. No significant PBDEs pollution was found in the areas.

[Determination of polybrominated diphenyl ethers in sewage sludge by high resolution gas chromatography coupled with high resolution mass spectrometry].

Polybrominated diphenyl ethers (PBDEs) are a group of brominated flame retardants, which are manufactured in large quantities and widely used in a variety of consumer goods. Recently they spread ubiquitously as environmental contaminants. In order to investigate the pollution level of PBDEs in different environmental samples, a method has been established by using high resolution gas chromatography coupled with high resolution mass spectrometry (HRGC-HRMS). The method was used to analyze biosamples obtained from the international inter-calibration. The results were satisfactory by analyzing z-score figures. Then, PBDE contents in three samples of sewage sludge were investigated by the method after Soxhlet extraction and multi-layer silica gel chromatographic separation. The number of PBDEs determined was 19. From the results, we can see that the contamination level of PBDEs in the sample from Beijing was higher than those from two other areas. The method was validated and applied for the analysis of environmental samples.

Quantitative structure-activity relationship models for prediction of the toxicity of polybrominated diphenyl ether congeners.

Levels of polybrominated diphenyl ethers (PBDEs) are increasing in the environment and may cause long-term health problems in humans. The similarity in the chemical structures of PBDEs and other halogenated aromatic pollutants hints on the possibility that they might share similar toxicological effects. In this work, three-dimensional quantitative structure activity relationships (3-D-QSAR) models, using comparative molecular field analysis (CoMFA) and comparative similarity indices analysis (CoMSIA), were built based on calculated structural indices and a reported experimental toxicology index (aryl hydrocarbon receptor relative binding affinities, RBA) of 18 PBDEs congeners, to determine the factors required for the RBA of these PBDEs. After performing leave-one-out cross-validation, satisfactory results were obtained with cross-validation O2 and R2 values of 0.580 and 0.995 by the CoMFA model and 0.680 and 0.982 by the CoMSIA model, respectively. The results showed clearly that the nonplanar conformations of PBDEs result in the lowest energy level and that the electrostatic index was the main factor reflecting the RBA of PBDEs. The two QSAR models were then used to predict the RBA value of 46 PBDEs for which experimental values are unavailable at present.