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Involvement of nitric oxide pathways in short term modulation of tyrosine hydroxylase activity by endothelins 1 and 3 in the rat anterior hypothalamus.

The ability of endothelins 1 and 3 (ET-1 and ET-3) to reduce neuronal norepinephrine release through ETB receptor activation involving nitric oxide (NO) pathways in the rat anterior hypothalamus region (AHR) was previously reported. In the present work, we studied the effects of ET-1 and -3 on tyrosine hydroxylase (TH) activity and the possible involvement of NO pathways. Results showed that ET-1 and -3 (10 nM) diminished TH activity in AHR and this effect was blocked by a selective ETB receptor antagonist (100 nM BQ-788), but not by a ET(A) receptor antagonist (BQ-610). To confirm these results, 1 microM IRL-1620 (ET(B) agonist) reduced TH activity whereas 300 nM sarafotoxin S6b falled to modify it. N(omega)-Nitro-L-arginine methyl ester (10 microM), 7-nitroindazole (10 microM), 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-ona (10 microM), KT5823 (2 microM), inhibitors of nitric oxide synthase, neuronal nitric oxide synthase, NO-sensitive-guanylyl cyclase, and protein kinase G, respectively, did not modify the reduction of TH activity produced by ETs. In addition, both 100 microM sodium nitroprusside and 50 microM 8-bromoguanosine-3',5'-cyclic monophosphate (NO donor and guanosine-3',5'-cyclic monophosphate analog, respectively) diminished TH activity. Present results showed that ET-1 and ET-3 diminished TH activity through the activation of ET(B) receptors involving the NO/guanosine-3',5'-cyclic monophosphate/protein kinase G pathway. Taken jointly present and previous results it can be concluded that both ETs play an important role as modulators of norepinephrine neurotransmission in the rat AHR.

Animals↗

Considerations about the enantioselective transformation of polycyclic musks in wastewater, treated wastewater and sewage sludge and analysis of their fate in a sequencing batch reactor plant.

The present work consists of two distinct parts: in the first part enantioselective GC was used to separate the different enantiomeric/diastereomeric polycyclic musks, PCMs (HHCB, AHTN, AHDI, ATII and DPMI) including the main transformation product of HHCB, HHCB-lactone, in wastewater and sewage sludge. After optimization all PCMs were resolved on a cyclodextrin containing Rt-BDEXcst capillary GC column. Enantiomeric ratios of PCMs in a technical mixture were determined and compared to those obtained from enantioselective separation of wastewater and sewage sludge samples. In general, enantiomeric ratios were similar for most materials in influent, effluent and stabilized sewage sludge. However, the ratios for HHCB, AHDI and particularly ATII suggest some stereospecific removal of these compounds. In the second part, a field study was conducted on a wastewater treatment plant comprising a sequencing batch reactor. Concentrations of HHCB, AHTN, ADBI, AHDI, ATII, DPMI and HHCB-lactone were determined by non-enantioselective GC in daily samples of influent, effluent and activated sludge during one week. Mean concentrations in influent were 6900 and 1520 ng/l for HHCB and AHTN, respectively. The other PCMs exhibited contents 200 ng/l. Mean percent removal was between 61% (AHDI) and 87% (HHCB) resulting in mean effluent concentrations below 860 ng/l. HHCB-lactone concentration increased during wastewater treatment with a mean in the influent of 430 ng/l and in the effluent of 900 ng/l, respectively, indicating a degradation of HHCB.

Benzopyrans↗

The anthropogenic contribution to the organic load of the Lippe River (Germany). Part I: Qualitative characterisation of low-molecular weight organic compounds.

GC/MS-screening analyses of water samples from the Lippe River, Germany, revealed the presence of a wide spectrum of low-molecular weight organic compounds ranging from non-polar constituents like aliphatic hydrocarbons to polar constituents like n-carboxylic acids and phenols. Most of the identified compounds could be attributed to anthropogenic input and are used as plasticizers, flame retardants, pharmaceutical drugs or fragrances. Some of them had rarely been noticed as organic pollutants of aquatic environments before. These are, among others, 9-methylacridine, the plasticizer 2,2,4-trimethyl-1,3-pentandioldiisobutyrate (TXIB), the surfactant 2,4,7,9-tetramethyl-5-decyne-4,7-diol (TMDD), triphenylphosphinoxide and the flame retardant tris(chloropropyl)phosphate. On the other hand, most of the identified trialkyl phosphates, pharmaceutical drugs and synthetic fragrances have been reported in surface waters by several authors so far. Input pathways of the detected compounds were traced back by sampling various input sources of organic matter such as discharges of wastewater and Lippe River tributaries. Several contaminants were ubiquitous in Lippe River water and also occurred in sewage effluent from a municipal sewage treatment plant and in samples from the tributaries. This observation suggests that they are typical sewage derived contaminants and have the potential to be used as anthropogenic molecular markers.

Drug Residues↗

Blood concentrations of polycyclic musks in healthy young adults.

Knowledge on the concentration of polycyclic musk fragrance compounds in human blood is sparse. This study examined the concentrations of six polycyclic musks in blood samples from healthy volunteers. Blood was taken from hundred healthy students of the Medical University of Vienna. The lipophilic fraction was extracted and after purification analyzed by GC-MS. Study participants also completed a questionnaire on the use of cosmetics, about nutrition and other life-style aspects. Two compounds -- galaxolide and tonalide -- were identified in higher percentages of the blood plasma samples. Maximum plasma levels over 100 ng/l were also only found for galaxolide (4100 ng/l) and tonalide (800 ng/l). Women showed significantly higher levels than men. In a statistical multivariate approach only use of body lotion and age were predictive of positive galaxolide concentrations. For tonalide no significant predictor could be found. The findings mirror the replacement of nitro musk fragrances by polycyclic musks, mainly galaxolide. The high concentrations of galaxolide in human blood raise concern since few toxicological data are available.

Adult↗

Polycyclic musk compounds in higher trophic level aquatic organisms and humans from the United States.

Polycyclic musks, 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta[g]-2-benzopyran (HHCB) and 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphthalene (AHTN), are used as fragrance ingredients in numerous consumer products such as cleaning agents and personal care products. Studies have reported the widespread occurrence of these musks in surface waters and fish from western European countries. Nevertheless, little is known about their accumulation in humans and wildlife in the United States. In this study, we measured concentrations of HHCB and AHTN in human adipose fat collected from New York City. Furthermore, tissues from marine mammals, water birds, and fish collected from US waters were analyzed to determine the concentrations of HHCB and AHTN. Concentrations of HHCB and AHTN in human adipose fat samples ranged from 12 to 798 and from <5 to 134 ng/g, on a lipid weight basis, respectively. A significant correlation existed between the concentrations of HHCB and AHTN in human adipose fat. Concentrations of HHCB and AHTN were not positively correlated with age or gender of the donors. HHCB was found in tissues of several wildlife species, but not in the livers of polar bear from the Alaskan Arctic. Among wildlife species analyzed, spinner and bottlenose dolphins collected from Florida coastal waters contained measurable concentrations of HHCB.

Adipose Tissue↗

Determination of polycyclic musks in sewage sludge from Guangdong, China using GC-EI-MS.

Polycyclic musks [Cashmeran (DPMI), Celestolide (ADBI), Phantolide (AHMI), Traseolide (ATII), Tonalide (AHTN) and Galaxolide (HHCB)] were Soxhlet extracted, separated and analyzed in sewage sludge from six different wastewater treatment plants from Guangdong, China, using GC and GC-EI-MS. DPMI, ADBI, HHCB, AHTN were found in all samples, and ATII was not found in any sample. AHMI was detected in five out of six samples. HHCB and AHTN were the two major polycyclic musks found in highest concentrations in sludge. The sludge from municipal wastewater treatment plants contained HHCB, AHTN and DPMI at concentrations between 5.416 and 21.214, 0.715 and 6.195 and 0.599 and 2.870 mg/kg (dry), respectively. The highest concentration was found in sludge from one cosmetic plant at 703.681 mg HHCB/kg (dry). The results indicate that there are two sources of polycyclic musks: domestic sewage and industrial wastewater.

Benzopyrans↗

The concentrations and distribution of polycyclic musks in a typical cosmetic plant.

Polycyclic musks [cashmeran (DPMI), celestolide (ADBI), phantolide (AHMI), traseolide (ATII), tonalide (AHTN) and galaxolide (HHCB)] in the air, wastewater, sludge samples of a typical cosmetic plant were analyzed. DPMI, ADBI, HHCB and AHTN were found in all samples, and ATII was not found in any sample. HHCB and AHTN were the major components in all samples. The polycyclic musk concentrations were very high in the air of the cosmetic plant, and polycyclic musks were mostly contained in the gas phase at the percentage of 86.35-97.70%. Average polycyclic musk concentrations in effluent were high, and ranged from 0.62 to 32.06 microgl-1. The removal efficiency during the active sludge wastewater treatment was also high, resulting from the adsorption of those compounds into the sludge. So the polycyclic musk concentrations were very high in the primary sludge and second sludge, and ranged from 1.78 to 92.45 mgkg-1 (dry), and from 2.87 to 65.67 mgkg-1 (dry), respectively. Results suggested that the sludge needed to be further treated to make polycyclic musks less influence to the environment.

Benzopyrans↗

Irrigation of treated wastewater in Braunschweig, Germany: an option to remove pharmaceuticals and musk fragrances.

In this study the fate of pharmaceuticals and personal care products which are irrigated on arable land with treated municipal wastewater was investigated. In Braunschweig, Germany, wastewater has been irrigated continuously for more than 45 years. In the winter time only the effluent of the sewage treatment plant (STP) of Braunschweig is used for irrigation, while during summer digested sludge is mixed with the effluent. In the present case study six wells and four lysimeters located in one of the irrigated agricultural fields were monitored with regard to the occurrence of 52 pharmaceuticals and two personal care products (PPCPs; e.g. betablockers, antibiotics, antiphlogistics, carbamazepine, musk fragrances, iodinated contrast media (ICM) and estrogens). No differences in PPCP pollution of the groundwater were found due to irrigation of STP effluents with and without addition of digested sludge, because many polar compounds do not sorb to sludge and lipophilic compounds are not mobile in the soil-aquifer. Most of the selected PPCPs were never detected in any of the lysimeter or groundwater samples, although they were present in the treated wastewater irrigated onto the fields. In the groundwater and lysimeter samples primarily the ICM diatrizoate and iopamidol, the antiepileptic carbamazepine and the antibiotic sulfamethoxazole were detected up to several mugl(-1), while the acidic pharmaceuticals, musk fragrances, estrogens and betablockers were likely sorbed or transformed while passing the top soil layer. Potential estrogenic effects are likely to disappear after irrigation, since the most potent steroid estrogens were not measurable.

Agriculture↗

Determination of fragrance allergens in cosmetics by size-exclusion chromatography followed by gas chromatography-mass spectrometry.

A method using size-exclusion chromatography (SEC) combined with gas chromatography-mass spectrometry (GC-MS) was developed for the quantitation of 24 restricted allergenic fragrance compounds in cosmetic samples. To achieve reproducible results fragrance calibration has to be performed with propyl acetate as a solvent containing a constant proportion of matrix components. With the exception of hydroxycitronellal (66+/-5%) all compounds showed good recovery rates in the range of 90-120%. The mean accuracy (relative error) was 1+/-10% for all 24 compounds in five spiked creams (10 mg/kg per allergen) and 8+/-34% in a reference sample (4-15 mg/kg). The biggest benefit compared to other methods is the flexible clean up with SEC which allows the determination of a large range of compounds in difficult matrices with GC-MS.

Allergens↗

Targeted multidimensional gas chromatography for the quantitative analysis of suspected allergens in fragrance products.

Two approaches are described and compared for the analysis of suspected allergens (SAs) in fragrance products, which are defined by the Scientific Committee of Cosmetics and Non-Food Products (SCCNFP). The first consists of a comprehensive two-dimensional gas chromatography (GC x GC) experiment using both a "conventional" non-polar/polar column combination and an "inverse" polar/non-polar column set. The second approach uses a targeted multidimensional gas chromatography (MDGC) system employing a Deans type pneumatic switch and a longitudinally modulated cryogenic system (LMCS). It was found that the conventional and inverse column sets complement each other well, providing identification of SAs present. Compounds well retained on the second dimension of one column set were the first to be eluted from the other. In some instances SAs co-eluting with matrix components on the second dimension for a given column set were clearly resolved on the other, although this has the disadvantage of requiring two analytical runs. Targeted MDGC with a non-polar/polar column set, successfully separated all SAs identified within a fragrance product. The instrument is set up in a similar fashion to a GC x GC system though with longer second dimension ((2)D) column, a cryogenic trap at the beginning of the second column, and a pneumatic switch coupling both columns. The data are easier to process than for a GC x GC experiment. The targeted MDGC method has the capacity to deliver far greater efficiency to targeted regions of a primary separation than a GC x GC experiment, whilst still maintaining overall run times similar to those of a conventional one-dimensional (1D) GC experiment. Cryogenic focussing at the beginning of the (2)D column delivers enhanced sensitivity, accurate (2)D retention times and narrow peak widths; these are responsible for an increased resolution obtained from the fast, relatively short (approximately 5 m) (2)D column. The two column set GC x GC analysis provided a quick and effective means to qualitatively determine the presence of six SAs in a commercially available air freshener, however all were not adequately resolved from matrix components. In contrast, quantitation was straightforward using the targeted MDGC method.

Allergens↗

Identification, quantitation and method validation for the analysis of suspected allergens in fragrances by comprehensive two-dimensional gas chromatography coupled with quadrupole mass spectrometry and with flame ionization detection.

The first part of this study first aims at validating a method to identify suspected allergens limited by EU regulations in fragrances by comprehensive two-dimensional gas chromatography (GC x GC) coupled with rapid scanning quadrupole MS (GC x GC-qMS). The effectiveness of the quadrupole MS operating at different scanning speed (1000 and 11,111 amu/s) was evaluated in identifying (full scan mode acquisition--TIC) and in quantifying (single ion monitoring--SIM) the target analytes in complex mixtures. In full scan acquisition mode the mass range was reduced to 40-240 amu to increase the scan acquisition rate while in SIM mode the influence of different dwell-times (40, 10 and 5 ms) was tested. The number of scans for each single modulated chromatographic GC x GC peak and the total number of scans for the 2D peak, together with half height peak width (referred to apex) of each allergen in the standard mixture in both TIC and SIM modes were determined. Moreover, the match quality of the spectra obtained by GC/MS at 1000 and 11,111 amu/s and by GC x GC-MS at 11,111 amu/s were compared and the occurrence of spectral skewing verified. In the second part of this work quantitative methods by GC x GC-SIM/qMS and GC x GC-FID were validated on the basis of Eurachem/CITAC protocols through which the following performance parameters were determined: confirmation of identity, selectivity and specificity, limit of detection (LOD), limit of quantitation (LOQ), linearity (working and linear range), precision and accuracy and uncertainty. Suspected allergens were spiked in a concentration range between 2 and 25 ppm (microg/mL) on a Test fragrance taken as a reference, while 1,4 dibromo-benzene and 4,4'-dibromodiphenyl were used as internal standards.

Allergens↗

Determination of retention indices in constant inlet pressure mode and conversion among different column temperature conditions in comprehensive two-dimensional gas chromatography.

A method to determine the second dimensional real retention time, dead times on both dimensions and retention indices in constant inlet pressure mode was developed in comprehensive two-dimensional gas chromatography. At the same time, the conversion of GC x GC retention indices among different column temperature conditions were also conducted based on some thermodynamics parameters. The calculation accuracies are better than 1.0 retention index unit. Furthermore, a retention index database was developed and used to identify the compounds in a cigarette essential oil sample. It showed that identification by the database was of close agreement with by time-of-flight mass spectrometry, and some isomers could also be distinguished based on the retention index database.

Allergens↗

Excretion and ecotoxicity of pharmaceutical and personal care products in the environment.

The presence and fate of pharmaceutical and personal care products (PPCPs) in the environment is undergoing increasing scrutiny. The existing clinical pharmacokinetics and pharmacodynamics data for 81 common compounds were examined for cues of ecotoxicity. Of these the proportions excreted were available for 60 compounds (i.e., 74%). The compounds had a low (< or =0.5%), a moderately low (6-39%), a relatively high (40-69%), or a high (> or =70%) proportion of the parent compound excreted. More than half of the compounds evaluated have low or moderately low proportions of the parent compound excreted. However, the proportions excreted were negatively but moderately correlated (r = -0.50; n = 13; P = 0.08) with the concentrations of the compounds in the aquatic environment, suggesting that the compounds that have low proportions excreted may also have inherently low degradability in the environment. Solubility, logK(ow), and pKa work well in predicting the behavior of PPCPs under clinical conditions and have been used in the environmental assessment of PPCPs prior to approval. However, these parameters did not correlate with the proportion of PPCPs excreted in the environment or their concentration in the environment, underscoring the need for research into the behavior of PPCPs in the environment. PPCPs occur in low concentrations in the environment and are unlikely to elicit acute toxicity. An ecotoxicity potential that is based on chronic toxicity, bioavailability, and duration of exposure to nontarget organisms is described as a guide in assessing the potency of these compounds in the environment.

Acetophenones↗

Evaluation of the dermal subchronic toxicity of phenoxyethyl isobutyrate in the rat.

Phenoxyethyl isobutyrate (PEIB) is a fragrance and food ingredient that has been granted GRAS status and approved by the FDA for food use. The present studies investigated the dermal absorption parameters and subchronic toxicity of PEIB. For the absorption, distribution and elimination study, Sprague-Dawley rats received a dermal application of 2-[ring U 14C]-PEIB under occlusion for 6 h. PEIB was diluted in diethyl phthalate (DEP) to administer, a total application volume of 2 ml/kg, concentrations of 0.5, 5 and 50% ( congruent with 10, 100 and 1000 mg PEIB/kgBW). Approximately 61-69% of the applied dose was recovered from the dressing and skin surface washing procedure performed after 6-h exposure. By 72 h post dose, systemic elimination of radioactivity was congruent with 18 to 19% of the absorbed dose via the urine with small amounts also found in the feces (<1.0%). Terminal (72 h) tissue analysis showed that 0.35-0.72% of the applied dose of radioactivity was retained in the carcass with low levels (</=0.03%) measured also in the liver, kidney and gastrointestinal tract. Plasma levels increased in a dose-related manner, with concentrations equal to 0.02, 0.2 and 2.0 microg equiv/ml from low to high dose, respectively. The total recovery for these studies ranged from 92.2 to 96.2% of the dermally applied radioisotope. In a 13-week subchronic rat toxicity study, daily dermal applications of PEIB were made under occlusion for 6 h. All groups were dosed at a constant 2 ml/kgBW volume of PEIB in the DEP vehicle at concentrations calculated to administer 0, 100, 300 or 1000 mg PEIB/kgBW/day. Clinical observations, assessments of skin irritation, hematology, and blood chemistry, necropsy, and gross and histopathologic evaluation of tissues demonstrated no treatment-related effects. The local skin irritation and systemic toxicity no-observed-effect-levels (NOELs) for PEIB in this study were determined to be >1000 mg/kgBW/day.

Administration, Cutaneous↗

Thresholds of elicitation depend on induction conditions. Could low level exposure induce sub-clinical allergic states that are only elicited under the severe conditions of clinical diagnosis?

While numerous studies have examined dose/response relationships occurring in the experimental induction of contact allergic dermatitis, fewer have examined the effects of varying the doses of both induction and challenge. Recently published studies have however done this and they all show the same remarkable observation: the threshold of elicitation decreases as the doses used to induce the allergy increase. This has important implications. One is that it may be more complicated to determine clear threshold doses below which allergic responses are not seen. It is also proposed that normal exposure to weak allergens such as some fragrance materials may induce "sub-clinical" allergic states which will not be elicited under these same exposure conditions but which may become apparent under the more severe conditions of clinical diagnosis. This may explain why the prevalence of Patch test reactions to some fragrance materials is apparently increasing in the absence of any clearly documented "epidemic" of consumer complaints.

Allergens↗