The discovery of new enzymes for poly(ethylene terephthalate) (PET) degradation has been a hot topic of research globally. Bis-(2-hydroxyethyl) terephthalate (BHET) is an intermediate compound in the degradation of PET and competes with PET for the substrate binding site of the PET-degrading enzyme, thereby inhibiting further degradation of PET. Discovery of new BHET degradation enzymes may contribute to improving the degradation efficiency of PET. In this paper, we discovered a hydrolase gene sle (ID: CP064192.1, 5085270-5086049) from Saccharothrix luteola, which can hydrolyze BHET into mono-(2-hydroxyethyl) terephthalate (MHET) and terephthalic acid (TPA). BHET hydrolase (Sle) was heterologously expressed in Escherichia coli using a recombinant plasmid, and the highest protein expression was achieved at a final concentration of 0.4 mmol/L of isopropyl-β-d-thiogalactoside (IPTG), an induction duration of 12 h and an induction temperature of 20 ℃. The recombinant Sle was purified by nickel affinity chromatography, anion exchange chromatography, and gel filtration chromatography, and its enzymatic properties were also characterized. The optimum temperature and pH of Sle were 35 ℃ and 8.0, and more than 80% of the enzyme activity could be maintained in the range of 25-35 ℃ and pH 7.0-9.0 and Co²⁺ could improve the enzyme activity. Sle belongs to the dienelactone hydrolase (DLH) superfamily and possesses the typical catalytic triad of the family, and the predicted catalytic sites are S129, D175, and H207. Finally, the enzyme was identified as a BHET degrading enzyme by high performance liquid chromatography (HPLC). This study provides a new enzyme resource for the efficient enzymatic degradation of PET plastics.
Actinomycetales/genetics* ; Hydrolases/metabolism* ; Phthalic Acids/chemistry* ; Polyethylene Terephthalates/metabolism*

OBJECTIVEConsuming phthalates may be due to the presence of food contact materials, such as plastic containers. In this study, we investigated the association between plastic container use and phthalate exposure in 2,140 Shanghai adults.

METHODSParticipants completed a questionnaire on the frequency of using plastic containers in different scenarios in the previous year (e.g., daily, weekly) and on the consumption of plastic-packaged foods in the previous three days (yes or no). Urinary phthalate metabolites were used to assess the association between phthalate exposure and the use of plastic containers.

RESULTSThe metabolites of di-(2-ethylhexyl) phthalate (DEHP) were the most frequently detected in urine. The results revealed that phthalate exposure was associated with consumption of plastic-packaged breakfast or processed food items in the previous three days. The consumption of these two food items had strong synergistic effects on increasing urinary concentrations of most phthalate metabolites.

CONCLUSIONOur results of plastic-packaged breakfast and processed food may be explained by the use of flexible plastic containers, indicating the importance of risk assessment for the application of flexible plastic containers.

Adult ; China ; Cities ; Data Collection ; Humans ; Phthalic Acids ; metabolism ; urine ; Plastics ; chemistry

The free membrane of Eudragit L100/S100 which is pH-sensitive, colon-specific was prepared by plane casting films. The film humidity, species and amount of plasticizers, the ratio of membrane material was investigated. The rate of membrane permeability and mechanical properties were used as indicators of orthogonal experiment, and its related properties were studied. The results show that the mechanical properties of the membrane and phragmoid capacity are the best when 30% TEC was used as plasticizer; the ratio of membrane material have little effect on the rate of membrane permeability and mechanical properties. By adjusting the species and amount of plasticizers, the ratio of Eudragit L100/S100, the free membrane which is colon-specific can be obtained.
Citrates ; chemistry ; Humidity ; Hydrogen-Ion Concentration ; Permeability ; Phthalic Acids ; chemistry ; Plasticizers ; chemistry ; Polymethacrylic Acids ; chemistry ; Technology, Pharmaceutical ; methods ; Tensile Strength ; Transition Temperature ; Triacetin ; chemistry

OBJECTIVETo evaluate whether mono (2-ethylhexyl) phthalate (MEHP) affects genomic DNA methylation and the methylation status of some specific genes such as patched gene (PTCH) and smoothened gene (SMO) in LNCaP cells.

METHODSLNCaP cells were treated with MEHP (0, 1, 5, 10, and 25 μmol/L) for 3 days. An ELISA assay was preformed to detect genomic methylation, including 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) content. A pyrosequencing assay was applied to assess DNA methylation in PTCH and SMO gene promoters. The correlation between DNA methylation and gene expression was assessed.

RESULTSThe proportion of cytosines with 5-mC methylation in LNCaP cells was significantly decreased by MEHP (1, 5, 10, and 25 μmol/L) in a dose-dependent manner (P < 0.01). For genes in the Hedgehog pathway, there was no significant MEHP concentration-dependent difference in the DNA methylation of PTCH and SMO.

CONCLUSIONMEHP might affect the progression of prostate cancer through its effect on global DNA methylation.

Antineoplastic Agents ; chemistry ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; Humans ; Male ; Phthalic Acids ; chemistry ; Prostatic Neoplasms ; metabolism

OBJECTIVE: To establish a scientific foundation for cosmetic supervision and administration based on the analysis of the sanitary quality of cosmetics in Hunan Province during 2010. METHODS: According to Cosmetic Sanitary Standards (set by the Ministry of Health, People's Republic of China), 150 random samples of cosmetics in Hunan were assayed both for microbial items (including total plate count, fungus and yeast, fecal coliform, staphylococcus aureus, pseudomonas aeruginosa) and chemical items (including 17 kinds of prohibited substances and 14 kinds of restricted substances). RESULTS: The total rate of cosmetics failing to meet the standards was 22.0% of the 150 samples; specific rates for failing perfumes, skin care products (eye cream) and deodorant products were, relatively, 70.6%, 60.00%, and 44.4%. Four kinds of prohibited substances, including diethyl phthalate, acrylamide, asbestos and neodymium, as well as 2 kinds of restricted substances, including triclosan and formaldehyde, were found to exceed standards. None of microbial items exceeded standard levels. CONCLUSION The sanitary quality control of cosmetics is lax. Administrative departments should not only reinforce their post-production supervision with respect to cosmetics, but also consolidate their control over the process of cosmetic production in order to solve the problem of toxic residues or illegal and intentional adulterations.
China ; Cosmetics ; analysis ; chemistry ; standards ; Formaldehyde ; isolation & purification ; Humans ; Phthalic Acids ; isolation & purification ; Quality Control ; Staphylococcus aureus ; isolation & purification

OBJECTIVETo study the dissolution rate of active components of different extracting solvents of Danggui Chishao drug pair.

METHODThe dissolution rates of phenolic acids (ferulic acid, vanillic acid and gallic acid), monoterpenes (gallic acid, peoniflorin, albiflorin, hydroxypeoniflorin and galloylpaeoniflorin) and phthalates (senkyunolide and ligustilide) contained in Danggui Chishao drug pair were determined by quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) and ultra high performance liquid chromatography in combination with triple-quadrupole mass spectrometry (UPLC-TQ-MS).

RESULTThe dissolution rates of phenolic acids and monoterpenes decreased with the increase in alcohol concentration, on the contrary the rates of phthalates increased. The relative dissolution rates of most active components were relatively high in water and low concentration alcohol than other solvents.

CONCLUSIONLiquid chromatography-mass spectrometry is practical for comprehensive multi-component assessment on traditional Chinese medicine preparation processes and can provide reference for optimization of processing parameters.

4-Butyrolactone ; analogs & derivatives ; chemistry ; Benzoates ; chemistry ; Benzofurans ; chemistry ; Bridged-Ring Compounds ; chemistry ; Chromatography, Liquid ; Drugs, Chinese Herbal ; chemistry ; Gallic Acid ; chemistry ; Glucosides ; chemistry ; Hydroxybenzoates ; chemistry ; Mass Spectrometry ; Monoterpenes ; chemistry ; Phthalic Acids ; chemistry ; Solvents ; chemistry ; Vanillic Acid ; chemistry

Combining the structural features of picotamide and linotroban, a series of N,N'-bis-(halogenophenyl)-4-methoxybenzene-1, 3-disulfonamides were designed and synthesized on the basic principles of drug design. The structures of target compounds were confirmed by IR, 1H NMR and HR-MS, and the in vitro antiplatelet aggregation activity was evaluated by Born turbidimetric method with adenosine diphosphate (ADP) as the platelet aggregation inducers. The assay results showed that twelve compounds (4b, 4f, 4l, 5b, 5d-5g, 5j, 5k, 5m and 5n) were found to have superior anti-platelet aggregation activities than the positive drug picotamide. The preliminary structure-activity relationship (SAR) has been explored.
Adenosine Diphosphate ; Drug Design ; Phthalic Acids ; Platelet Aggregation ; Platelet Aggregation Inhibitors ; chemical synthesis ; chemistry ; Structure-Activity Relationship ; Sulfonamides ; chemical synthesis ; chemistry

OBJECTIVETo study the changes of the fatty components of Pollen Typhae before and after being carbonized.

METHODPollen Typhae and Pollen Typhae carbonisatus were extracted with petroleum ether (60-90 degrees C) respectively. The two kinds of extracts were analyzed by GC-MS after saponificated and methanolized, and their constituents were searched through NIST. The contents of the constituents were determined by method of normalization.

RESULTEither in Pollen Typhae or in Pollen Typhae carbonisatus, 32 components were identified, among which 20 components were the same and 6 were different respectively. Among the same components, the relative contents of 3-methyl-2-butenoic acid-2-phenylethyl ester, hexanedioic acid-dimethyl ester, dimethyl phthalate, diethyl phthalate, diphenylamine, sebacic acid dimethyl ester, 1,2-benzenedicarboxylic acid, ethyl methyl ester, methyl-2-ethylhexyl phthalate and diisooctyl phthalate etc. increased obviously, and the relative contents of nonanedioic acid-dimethyl ester, diisobutyl phthalate and stigmastan-3,5-dien etc. decreased greatly. Among the different components, 8-hydroxy-octanoic acid-methyl ester, 9-hydroxy-nonanoic acid-methyl ester, 10-octadecenoic acid-methyl ester, m-hydroxycinnamic acid-methyl ester,3-[4-( acetyloxy)-3-methoxyphenyl]2-propenoic acid-methyl ester and 11-octadecenoic acid-methyl ester were detected in Pollen Typhae, 3-hydroxyspirost-8-en-11-one, benzenepropanoic acid-methyl ester, 2,4-dimethylhexanedioic acid; 2,4-bis (1,1-dimethylethyl)-phenol, undecanedioic acid-dimethyl ester and 9,10-dihydroxy-octadecanoic acid-methyl ester were detected in Pollen Typhae carbonistatus.

CONCLUSIONThe species and contents of the fatty components in Pollen Typhae changed before and after being carbonized, but their chemical types didn't change too much.

Carbon ; Dibutyl Phthalate ; analogs & derivatives ; analysis ; Fatty Acids ; analysis ; chemistry ; Gas Chromatography-Mass Spectrometry ; Hot Temperature ; Phthalic Acids ; analysis ; Plants, Medicinal ; chemistry ; Pollen ; chemistry ; Technology, Pharmaceutical ; methods ; Typhaceae ; chemistry

AIMTo analyze the chemical components in Danggui (the roots of Angelica sinensis (Oliv.) Diel).

METHODSHPLC-MS/MS was used to identify the main components in Danggui. Furthermore, the MS fragmentation regularity of the phthalides was proposed. The mobile phase of HPLC consisted of 0.5% acetic acid in water and 0.5% acetic acid in acetonitrile, analytical column was Hypersil ODS2 (250 mm x 4.6 mm, 5 microm), flow rate 1.0 mL x min(-1), injected volume 2 microL. The ionization source was ESI in positive ion mode.

RESULTSFerulic acid, nine known phthalides and one unknown phthalide derivative were tentatively identified in chromatograms based on their MS data and the comparison of their UV spectra with those published in the literatures.

CONCLUSIONThe structural information of phthalides was obtained via HPLC-MS/MS, which provides an accurate and fast method to identify the phthalides and provides more scientific information for quality control of Danggui.

4-Butyrolactone ; analogs & derivatives ; analysis ; chemistry ; Angelica sinensis ; chemistry ; Benzofurans ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; analysis ; chemistry ; Molecular Structure ; Phthalic Anhydrides ; analysis ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Spectrometry, Mass, Electrospray Ionization ; methods

The concentrations of 16 phthalates in 164 commercial Chinese rice wines (CRW) were detected by GC-MS, and consumption data on CRW in different packaging types was investigated from 634 adult males in Shanghai using a food frequency questionnaire. Based on the principles of probabilistic modelling and cumulative risk assessment, the exposure and health risk of phthalates from CRW to adult males in Shanghai was evaluated. DMP, DEP, DIBP, DnBP, BBP, and DEHP were detected in the samples, the range of detection frequency of individual phthalates varied from 6.10% for BBP to 15.24% for DIBP, and the detected concentrations were 51.06-200.34 ng/mL. All the respondents consumed CRW, 90.69% of them consumed CRW 0.01-49.9 mL/d, the minimum value of the average daily intake of CRW was 6.25 mL/d, the median was 13.72 mL/d and the maximum was 300 mL/d. The median exposure level of the 6 detected Phthalates to adult males in Shanghai were 6.58-7.10 ng/(d•kg), and the maximum exposure level were 137.38-540.47 ng/(d•kg). The cumulative exposure health risk index (HI) based on the median and maximum exposure level of the 6 Phthalates (DMP, DEP, DIBP, DnBP, BBP, and DEHP) were 0.001147 and 0.063396, both were far less than 1. In conclusion, CRW were generally consumed by the adult males in Shanghai, although multiple phthalates were detected in commercial CRW, health risk of such exposure levels from commercial CRW to the target adult males in Shanghai was very low.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; Cities ; Environmental Exposure ; Environmental Pollutants ; analysis ; Food Safety ; Humans ; Male ; Middle Aged ; Oryza ; chemistry ; Phthalic Acids ; analysis ; Risk Assessment ; Wine ; analysis ; Young Adult