Unlocking the Therapeutic Potential of Freshwater Sapropel Extracts: In Vitro Analysis and Antioxidant Profiling for Skincare Applications.

Background and Objective: Sapropel, a biologically active sedimentary deposit, is high in organic matter and minerals and has been shown to offer health benefits. Its constituents, humic acid (HA) and fulvic acid (FA), have been found to have some therapeutic applications. The aim of this study was to determine the potential therapeutically significant properties of freshwater sapropel extracts: their polyacid content, antioxidative (AO) status, and biological activity in cell culture. Materials and Methods: Freshwater lakes from the southeast region of Latvia were investigated layer by layer. The total organic carbon (TOC) was determined through combustion using the catalytic oxidation method, HA and FA were measured via acid perspiration, and the total polyphenol content (TPC) and total antioxidant status (TAS) was analysed spectrophotometrically. Sapropel extracts' regenerative abilities were tested in vitro using a Cell-IQ real-time monitoring system on mouse BALB/c 3T3 fibroblasts and human keratinocyte HaCaT cell lines. Cytotoxicity was measured through neutral red uptake assessment as a concentration-dependent reduction in the uptake of neutral red dye relative to a vehicle control or untreated cells. Results: The highest AO activity was observed in sapropel extracts with elevated concentrations of HA and TPC from Audzelu Lake (1.08 ± 0.03 mmol/L), and the lowest activity was found in extracts from Ivusku Lake (0.31 ± 0.01 mmol/L). Correspondingly, the concentrations of HA in Audzelu and Ivusku Lakes were recorded as 45.2 and 27.4 mg/g, respectively. High concentrations of HA promoted in vitro cell growth upon short-term exposure (up to 6 h). Conclusions: The results show that high TPC correlates with AO status and sapropel extracts with higher concentrations of HA exhibit greater AO activity and promote in vitro cell growth, suggesting a perspective use for short-term topical therapeutic skin applications. However, higher concentrations over longer durations showed cytotoxic effects, indicating the need for further investigation.

Assessing Contributions of Synthetic Musk Compounds from Wastewater Treatment Plants to Atmospheric and Aquatic Environments.

The high environmental concentrations, persistence, and toxicity of synthetic musk compounds (SMCs) necessitate a better grasp of their fate in wastewater treatment plants (WWTPs). To investigate the importance of WWTPs as pathways of SMCs to the environment, air and wastewater samples were collected at four WWTPs in Ontario, Canada. Polycyclic musks (PCMs) were present at higher concentrations than nitro musks (NMs) and macrocyclic musks (MCMs). Three PCMs [galaxolide (HHCB), tonalide (AHTN), and iso-E super (OTNE)] were the most abundant compounds (0.30-680 ng/m3 in air, 0.40-15 µg/L in influent, and 0.007-6.0 µg/L in effluent). Analyses of multiyear data suggest that risk management measures put in place have been effective in reducing the release of many SMCs into the environment. The highest removal efficiency, up to almost 100% of some SMCs, was observed for the plant with the longest solid retention time. A fugacity-based model was established to simulate the transport and fate of SMCs in the WWTP, and good agreement was obtained between the measured and modeled values. These findings indicate that the levels of certain SMCs discharged into the atmospheric and aquatic environments were substantial, potentially resulting in exposure to both humans and wildlife.

Glacier foreland insect uptake synthetic compounds: an emerging environmental concern.

Pesticides, synthetic fragrances and polycyclic aromatic hydrocarbons contaminated two glacier-fed streams (Amola, Mandrone) and one spring (Grostè) in the Italian Alps. Ten compounds (chlorpyrifos (CPY), chlorpyrifos-methyl (CPY-m), galaxolide (HHCB), tonalide (AHTN), fluorene (Flu), phenanthrene (Phen), anthracene (Ant), fluoranthene (Fl), pyrene (Pyr), benzo[a]anthracene (BaA)) accumulated in aquatic larvae of chironomids (Diamesa steinboecki, D. latitarsis, D. bertrami, D. tonsa, D. zernyi, Pseudokiefferiella parva, Orthocladiinae) and tipulids. Their tissue concentrations (detected by gas chromatography coupled with mass spectrometry) ranged from 1.1 ± 0.1 ng/g d.w. (= dry weight) (CPY-m in D. tonsa from Amola) to 68.0 ± 9.1 ng/g d.w. (Pyr in D. steinboecki from Mandrone). HHCB, AHTN, and CPY, with one exception, were accumulated by all aquatic insects. Six compounds (CPY, CPY-m, HHCB, AHTN, Fl, Pyr) also contaminated carabids (Nebria germarii, N. castanea, N. jockischii) predating adults of merolimnic insects. Their tissue concentrations ranged from 1.1 ± 0.3 ng/g d.w. (CPY-m in N. germarii from Mandrone) to 84.6 ± 0.3 ng/g d.w. (HHCB in N. castanea from Grostè). HHCB and AHTN were accumulated by all Nebria species. Intersite and interspecies differences were observed, which might be attributed to different environmental contamination levels. There was a stronger similarity between species from the same site than among the same species from different sites, suggesting that uptake is not species specific. At all sites, the concentration of xenobiotics was higher in larvae than in water and comparable or higher in carabids than in larvae from the same site, suggesting trophic transfer by emerging aquatic insects to their riparian predators.

Ecological risk of galaxolide and its transformation product galaxolidone: evidence from the literature and a case study in Guangzhou waterways.

Galaxolidone (HHCB-lac) is a major transformation product of the commonly used synthetic musk galaxolide (HHCB) and is ubiquitous in the environment along with the parent compound. Although many studies have shown the harmful effects of HHCB, little attention has been paid to the potential ecological risk of HHCB-lac. Herein, we reviewed the concentrations and ratios of HHCB and HHCB-lac (HHCB-lac : HHCB) in different media reported in the literature, derived the predicted no-effect concentrations (PNECs) for the two compounds using ECOSAR predictions and species sensitivity distribution (SSD) estimates, and assessed their ecological risks in the aquatic environment. The literature data indicated that HHCB-lac and HHCB were generally present in the environment at ratios of 0.01-10. Using the derived PNECs (2.14 and 18.4 µg L-1 for HHCB and HHCB-lac, respectively), HHCB in the aquatic environment was assessed to have medium to high risks, while HHCB-lac was assessed to have low risks. Furthermore, we carried out a case study on the occurrence and ecological risks of HHCB and HHCB-lac in Guangzhou waterways. The concentrations of the two compounds in Guangzhou waterways ranged from 20 to 2620 ng L-1 and 3 to 740 ng L-1, respectively, and the ratios were in the range of 0.15 to 0.64. The field study data also showed medium to high risks of HHCB and low risks of HHCB-lac. Additionally, the endocrine effects of HHCB and HHCB-lac were confirmed by Endocrine Disruptome, which calls for greater scrutiny of the potential effects of HHCB and HHCB-lac on human health.

Global distribution and ecological risk assessment of synthetic musks in the environment.

Synthetic musks, as an alternative product of natural musks, are widely used in almost all fragrances of consumer products, such as perfumes, cosmetics and detergents. During the past few decades, the production of synthetic musks has been increasing year by year, subsequently followed by large concern about their adverse effects on ecosystems and human beings. Until now, several studies have reviewed the latest development of analytical methods of synthetic musks in biological samples and cosmetics products, while there is still lack of a systematic analysis of their global distribution in different environmental media. Thus, this review summarizes the occurrence of synthetic musks in the environment including biota around the world and explores their global distribution patterns. The results show that galaxolide (HHCB), tonalide (AHTN), musk xylene (MX) and musk ketone (MK) are generally the most frequently detected synthetic musks in different samples with HHCB and AHTN being predominant. Higher concentrations of HHCB and AHTN are normally found in western countries compared to Asian countries, indicating more consumptions of these musks in western countries. The persistence, bioaccumulation and toxicity (PBT) of synthetic musks (mainly for polycyclic musks and nitro musks) are also discussed. The risk quotients (RQs) of HHCB, AHTN, MX and MK in most waters and sediments are below 0.1, reflecting a low risk to aqueous and sediment-dwelling species. In some sites, e.g., close to STPs, high risks (RQs>1) are characterized. Currently, limited data are available for macrocyclic musks and alicyclic musks in terms of either occurrence or PBT properties. More studies with an expanded scope of chemical type, geographical distribution and (synergic) toxicological effects especially from a long-term point of view are needed.

Bioremediation of the synthetic musk compounds Galaxolide and Tonalide by white rot fungal strain-assisted phytoremediation in biosolid-amended soil.

The study was aimed to conduct the bioremediation of synthetic musks by four species of white rot fungi combined with phytoremediation (Zea mays) in biosolid-amended soils where only Galaxolide (HHCB) and Tonalide (AHTN) were found as other musks were below the detection limit (0.5-2 µg/kg dw). The HHCB and AHTN concentration in natural attenuation treated soil was decreased by not more than 9%. In solely mycoremediation, Pleurotus ostreatus was found to be the most efficient fungal strain, with the higher (P < 0.05) HHCB and AHTN removal (51.3% and 46.4%). Phytoremediation-only of biosolid-amended soil was also able to remove HHCB and AHTN from soil significantly (P < 0.05) in comparison to the control treatment without plants which resulted in the final concentration for both compounds of 56.2 and 15.3 µg/kg dw, respectively. Using white rot fungus-assisted phytoremediation, only P. ostreatus decreased the HHCB content in soil significantly (P < 0.05) by 44.7%, when compared to the initial concentration. While using Phanerochaete chrysosporium, the AHTN concentration was decreased by 34.5%, which was a significantly lower concentration at the end of experiment compared to the initial value. Via fungus-assisted phytoremediation, the enzymatic activity and fungal biomass were increased, probably due to the presence of roots in association with the soil microbiome, in the process increasing the degradation of fragrances accordingly. This could lead to a higher (P < 0.05) AHTN removal in P. chrysosporium assisted phytoremediation. Estimated HHCB and AHTN bioaccumulation factors in maize were lower than 1, therefore no environmental risk would be posed.

The impact of synthetic musk compounds in biofilms from drinking water bacteria.

Musk fragrances have been detected in drinking water (DW) at trace concentrations. However, their impact on the microbial quality of DW has been disregarded. This work provides a pioneer evaluation of the effects of two synthetic musks contaminants, tonalide (AHTN) and galaxolide (HHCB), in microbial biofilms formed on two different surfaces, polyvinyl chloride (PVC) and stainless steel AISI 316 (SS316). Three bacterial species isolated from DW (Acinetobacter calcoaceticus, Burkholderia cepacia and Stenotrophomonas maltophilia), were used to develop 7-day-old single and mixed species biofilms. The impact of musks was assessed directly on biofilms but also on the bacteria motility, biofilm formation ability and biofilm susceptibility to chlorination. AHTN musk caused the most remarkable effects by increasing the cellular density and viability of mixed biofilms, and the extracellular polysaccharides content of biofilms on SS316. Most of the alterations caused by the direct exposure of biofilms to musks were observed when SS316 was used as an adhesion surface. In contrast, the ability to form biofilms and their susceptibility to chlorine were more affected for bacteria from HHCB-exposed biofilms on PVC. The overall results demonstrate that the presence of musks at residual concentrations influences DW bacterial dynamics, with the potential to impact the DW quality and safety. The type of plumbing material may further impact the effects of musks.

Metabolites in Conocarpus erectus leaves attenuate α-amylase activity by modulating amino acid residues of α-amylase: an in vitro and docking study / Los metabolitos en las hojas de Conocarpus erectus atenúan la actividad de la α-amilasa al modular los residuos de aminoácidos de la α-amilasa: un estudio in vitro y de acoplamiento

The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 ́-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.

Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 ́-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.

Using a fugacity model to determine the degradation rate of typical polycyclic musks in the field: A case study in the North Canal River watershed of Beijing, China.

To quantitate the degradation rate of 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) under field conditions, a level III fugacity model combined with a least-squares method was used to determine the degradation rate of HHCB and AHTN in the North Canal River watershed of Beijing, China. Model fitting, validation, sensitivity, and uncertainty analyses revealed that the established model was stable and robust. The degradation rates of HHCB and AHTN were 4.16 × 10-3 h-1 (t1/2 = 167 h) and 1.68 × 10-2 h-1 (t1/2 = 41.3 h), respectively. The calculated degradation rates were extrapolated to the Liangshui River, and indicated that the differences between the measured and predicted concentrations were less than 0.32 and 0.34 log units for HHCB and AHTN, respectively. The attenuation rates of HHCB and AHTN were calculated, and the results indicated that degradation was an important yet not the sole contributor to the degradation of the polycyclic musks. Results of uncertainty analyses indicated that the inflow and outflow concentrations of the polycyclic musks in the surface water of each segment strongly influenced the model outputs, followed by environmental factors (water depth and flow rate). It is essential to measure the degradation rate in the field because of the influence of the surrounding environment. The present study reveals the utility of fugacity models to quantify the degradation rate of organic micropollutants in the field.

Sonoelectrochemical activation of peroxymonosulfate: Influencing factors and mechanism of FA degradation, and application on landfill leachate treatment.

In this work, sonoelectrochemically activated peroxymonosulfate (US-EC/PMS) was used to degrade fulvic acid (FA) in water. Compared with other technologies, the US-EC/PMS system can achieve higher FA decolorization in a short time. Moreover, the benefits of synergy are more prominent in the US-EC/PMS system. The effects of operating parameters on the sonoelectrochemical degradation of FA were investigated, including initial pH, initial FA concentration, current density, ultrasonic power, PMS dosage. The results showed the initial FA concentration and current density were critical to the degradation of FA. Under optimized parameters: initial pH of 2, 50 mg L-1 initial FA concentration, 30 mA cm-2 current density, 50 W ultrasonic power, 1 mM PMS dosage, the US-EC/PMS system can achieve 93% FA decolorization. The calculation results of current efficiency and energy consumption indicate that the introduction of PMS into the US-EC system has economic applicability. Scavenger experiments and electron paramagnetic resonance suggest that hydroxyl radicals, sulfate radicals, and singlet oxygen were the main ROS produced in the US-EC/PMS system. Accordingly, the possible mechanism of FA degradation by sonoelectrochemical activation PMS was proposed. Finally, the US-EC/PMS system was used to treat the aged landfill leachate. Three-dimensional fluorescence analysis showed that most of the humic substances (Hss) were effectively removed, and the biodegradability of the leachate was considerably improved. In addition, the effective removal of COD, chroma, and ammonia nitrogen were observed, proving that this technology is a powerful means to treat organic wastewater contaminated by Hss.

RIFM fragrance ingredient safety assessment, 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel-, CAS Registry Number 943723-15-7.

The existing information supports the use of this material as described in this safety assessment. 5,8-Methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, phototoxicity/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analog 5,8-methano-2H-1-benzopyran-2-one, 6- ethylideneoctahydro- (CAS # 69486-14-2) show that 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- is not expected to be genotoxic. The repeated dose, reproductive, and local respiratory toxicity endpoints were evaluated using the TTC for a Cramer Class III material, and the exposure to 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- is below the TTC (0.0015 mg/kg/day, 0.0015 mg/kg/day, and 0.47 mg/day, respectively). Data provided 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- a NESIL of 8200 µg/cm2 for the skin sensitization endpoint. The phototoxicity/photoallergenicity endpoints were evaluated based on human study data and UV/Vis spectra; 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- is not expected to be phototoxic/photoallergenic. The environmental endpoints were evaluated; 5,8-methano-2H-1-benzopyran, 6(or 7)-ethylideneoctahydro-, [4aR,5S,8S,8aS(or 4aR,5R,8S,8aR)]-rel- was found not to be PBT as per the IFRA Environmental Standards, and its risk quotients, based on its current volume of use in Europe and North America (i.e., PEC/PNEC), are <1.

Occurrence, spatial distribution, and fate of polycyclic musks in sediments from the catchment of Chaohu Lake, China.

Twenty-nine surface sediments from Chaohu Lake in China and from its six main tributaries were sampled to investigate the concentrations of two important polycyclic musks (PCMs), 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta(g)-2-benzopyran (galaxolide, HHCB) and 7-acetyl-1,1,3,4,4,6-hexamethyl-1,2,3,4-tetrahydronaphthalene (tonalide, AHTN), as well as the concentration of 4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta(g)-2-benzopyran-1-one (galaxolidon, HHCB-lactone), which is the main degradation product of HHCB. Except for the high concentrations of AHTN and HHCB measured in the Nanfei River (879 ng/g dw and 5,513 ng/g dw, respectively), the levels of AHTN and HHCB in the river sediments were 7.08-44.9 ng/g dw and 20.6-268 ng/g dw, respectively, which are slightly lower than those documented in various areas worldwide. The concentrations of AHTN and HHCB in the sediments of Chaohu Lake were one or two orders of magnitude lower than those in the tributary rivers and showed a clear regional distribution. The concentrations of HHCB-lactone were comparable to those of HHCB and presented a significant positive correlation with the concentrations of HHCB, suggesting that the HHCB-lactone originated directly from the degradation of HHCB in wastewater treatment plants (WWTPs) or in the natural environment. The diagnostic ratios of HHCB/AHTN and HHCB-lactone/HHCB and the enantiomeric fractions (EFs) of these PCMs showed that the direct origins of the target PCMs in the study area were municipal and industrial wastewaters discharged from adjacent cities or point sources and that the HHCB-lactone in sediment originated from the natural degradation of HHCB in the rivers and the lake. The results of the risk assessment showed that the PCMs in the watershed sediments were unlikely to pose a threat to aquatic species. However, the effluents of industrial and municipal wastewaters that are discharged into the Nanfei River should be investigated in future research.

3D-QSAR-aided toxicity assessment of synthetic musks and their transformation by-products.

Synthetic musks (SMs) are fragrance additives widely used in personal care products. SMs and their transformation by-products may reach the environment even after wastewater treatment, resulting in ecological and health concerns. The identification and toxicity assessment of SM by-products generated from different chemical and biological treatment processes have been rarely studied. This study established a 3D-QSAR model based on SMs' molecular structures (independent variable) and their lethal concentration (LC50) of mysid (dependent variable). The developed model was further used to predict the LC50 of SMs transformation by-products. Fifty-eight by-products of six common SMs (i.e., galaxolide (HHCB), tonalide (AHTN), phantolide (PHAN), traseolide (TRASE), celestolide (ADBI), and musk ketone (MK)) generated from biodegradation, photodegradation, advanced oxidation, and chlorination were identified through literature review and lab experiment as the model inputs. Predicted LC50 results indicated that the toxicity of 40% chlorination by-products is higher than their precursors. Biodegradation is an effective method to treat AHTN. The advanced oxidation may be the best way to treat HHCB. This is the first study on biotoxicity of SM transformation by-products predicted by the 3D-QSAR model. The research outputs helped to provide valuable reference data and guidance to improve management of SMs and other emerging contaminants.

Determination of Organic Compounds, Fulvic Acid, Humic Acid, and Humin in Peat and Sapropel Alkaline Extracts.

Black, brown, and light peat and sapropel were analyzed as natural sources of organic and humic substances. These specific substances are applicable in industry, agriculture, the environment, and biomedicine with well-known and novel approaches. Analysis of the organic compounds fulvic acid, humic acid, and humin in different peat and sapropel extracts from Lithuania was performed in this study. The dominant organic compound was bis(tert-butyldimethylsilyl) carbonate, which varied from 6.90% to 25.68% in peat extracts. The highest mass fraction of malonic acid amide was in the sapropel extract; it varied from 12.44% to 26.84%. Significant amounts of acetohydroxamic, lactic, and glycolic acid derivatives were identified in peat and sapropel extracts. Comparing the two extraction methods, it was concluded that active maceration was more efficient than ultrasound extraction in yielding higher amounts of organic compounds. The highest amounts of fulvic acid (1%) and humic acid and humin (15.3%) were determined in pure brown peat samples. This research on humic substances is useful to characterize the peat of different origins, to develop possible aspects of standardization, and to describe potential of the chemical constituents.

Calibration and measurement of mitochondrial pH in intact adult rat cardiomyocytes.

Mitochondrial pH is a vital parameter of the mitochondrial environment, which determines the rate of many mitochondrial functions, including metabolism, membrane potential, fate, etc. Abnormal mitochondrial pH is always closely related to the health status of cells. Analyzing mitochondrial pH can serve as a proxy for mitochondrial and cellular function. This protocol describes the use of SNARF-1 AM, a pH-sensitive fluorophore, to measure mitochondrial pH. This protocol details the steps to evaluate mitochondrial pH in live adult cardiomyocytes using confocal microscopy. The protocol can be adapted to other adherent cell types. For complete details on the use and execution of this protocol, please refer to Wei-LaPierre et al. (2013).

HPLC-UV/HRMS methods for the unambiguous detection of adulterations of Ginkgo biloba leaves with Sophora japonica fruits on an extract level.

CONTEXT: Ginkgo biloba L. (Ginkgoaceae) leaf extract is one of the most frequently sold herbal extracts. There have been reports on poor quality and adulteration of ginkgo leaf extracts or the powdered plant material with extracts or powder of Styphnolobium japonicum (L.) Schott (Fabaceae) (syn. Sophora japonica L.) fruits, which is rich in flavone glycosides. OBJECTIVE: The study investigates whether ginkgo leaves genuinely contain genistein and sophoricoside and whether these two substances could be used as markers to detect adulterations with sophora fruits. MATERIALS AND METHODS: A total of 33 samples of dried ginkgo leaves were sourced from controlled plantations in China, the USA, and France. After extraction, the samples were analyzed using two high-performance liquid chromatography (HPLC) coupled with UV/HRMS methods for the detection of genistein and sophoricoside, respectively. Chromatograms were compared to standard reference materials. RESULTS: In none of the tested ginkgo samples, neither genistein nor sophoricoside could be detected. The applied method was designed to separate genistein from apigenin. The latter is a genuine compound of ginkgo leaves, and its peak may have been previously misidentified as genistein because of the same molecular mass. The method for the detection of sophoricoside allows identification of the adulteration with sophora fruit without prior hydrolysis. By both HPLC methods, it was possible to detect adulterations of ≥2% sophora fruits in the investigated ginkgo extract. CONCLUSION: The methods allow unambiguous detection of adulterations of ginkgo leaves with sophora fruits, using genistein and sophoricoside as marker compounds.

Large-scale preparative isolation of bergenin standard substance from Saxifraga atrata using polyamide coupled with MCI GEL® CHP20P as stationary phases in medium pressure chromatography.

In this study, polyamide and MCI GEL® CHP20P were employed as stationary phases in medium pressure chromatography (MPC) for the efficient preparative separation of bergenin from Saxifraga atrata. Ethanol-water, methanol-water, and acetonitrile-water mobile phases all showed good enrichment capacity for bergenin fraction when polyamide was used as a stationary phase. After 5 cycles of polyamide MPC using acetonitrile/water, 1.2 g of bergenin fraction was isolated from 180 g Saxifraga atrata herb. Further purification of this fraction was conducted using MCI GEL® CHP20P styrene-divinylbenzene beads. The bergenin fraction was separated into two fractions, and after three runs of MPC, 714.2 mg of bergenin with purity above 99% was obtained. The results demonstrate that the combination of polyamide and styrene-divinylbenzene MPC can be utilized for preparative isolation of compounds from natural products with high yield and purity.

Polycyclic musk fragrance (PMF) removal, adsorption and biodegradation in a conventional activated sludge wastewater treatment plant in Northern Italy.

The first Italian annual monitoring study was carried out in Northern Italy to analyse the fate and removal of polycyclic musk fragrances (PMFs) in a wastewater treatment plant (WWTP) with conventional activated sludge (CAS) system. Water was sampled in four different stations along wastewater treatments to better understand the behaviour of PMFs along different steps of the plant. Galaxolide (HHCB) and galaxolidone (HHCB-lactone) were found in concentrations at µg L-1 level, 1 order of magnitude greater than tonalide (AHTN), whilst phantolide (AHDI) was never detected and celestolide (ADBI) was measured only at trace levels. Considering water concentrations, HHCB and AHTN evidenced a slight reduction, 20% and 50%, respectively, during wastewater treatments, thus resulting in a modest removal efficiency, mainly due to adsorption processes during the biological treatment. This was also confirmed by the high PMF concentrations measured in activated sludges which remained stable throughout the year. On the contrary, HHCB-lactone registered an increase up to 70% during wastewater treatments caused by the biotransformation of the parental compound HHCB during the biological treatment, as shown by the different HHCB-lactone/HHCB ratio measured before and after this step. No significant differences were recorded between seasons in terms of PMF input onto WWTP, in accordance with the common use of these chemicals in civil houses. Overall, this study suggests that current technologies employed in conventional WWTP are not enough efficient in removing these organic micropollutants from wastewaters and, therefore, WWTP effluents represent possible point sources of pollution for aquatic ecosystems. Additional treatments are necessary to enhance the removal of PMFs in order to increase the quality of the WWTP effluents.

Levels and trends of synthetic musks in marine bivalves from French coastal areas.

The levels and trends of four bioaccumulative synthetic musks (galaxolide - HHCB, tonalide - AHTN, musk xylene - MX and musk ketone - MK) were investigated in filter-feeding bivalves collected yearly since 2010 at sites of contrasted pressure along the French coasts. Quantification rates were high for all 4 compounds (85-99%), indicating their geographical and temporal extensive occurrence in the French coastal environment. The polycyclic musks HHCB and AHTN prevailed, with median concentrations of 2.27 ng g-1 dw and of 0.724 ng g-1 dw, whilst nitromusks were found 1 to 2 orders of magnitude lower. These levels were in the high range of those encountered for various other CEC families at the same sites and comparable to those from other locations on European coasts. Unlike for the other musks, the accumulation of HHCB was evidenced to be species-specific, with significantly lower levels found in oysters in comparison with mussels, possibly suggesting a higher metabolization in oysters. Geographical differences in musk distribution highlighted the sites under strong anthropogenic pressures and these differences were found to be consistent between years. The HHCB/AHTN ratio proved to be discriminant to explain the relative occurrence of polycyclic musks. The 8-year time series showed that only the now-banned compound MX displayed a significant decrease in most sites, whilst stable concentrations of the other musks suggested consistency in their usage over the last decade. These results provide reference data for future studies of the occurrence of personal care products on European coasts.

HSCCC Separation of Three Main Compounds from the Crude Extract of Dracocephalum Tanguticum by Using Dimethyl Sulfoxide as Cosolvent.

Separation of natural compounds directly from the crude extract is a challenging work for traditional column chromatography. In the present study, an efficient method for separation of three main compounds from the crude extract of Dracocephalum tanguticum has been successfully established by high-speed counter-current chromatography (HSCCC). The crude extract was directly introduced into HSCCC by using dimethyl sulfoxide as cosolvent. Ethyl acetate/n-butyl alcohol/0.3% glacial acetic acid (4: 1: 5, v/v) system was used and three target compounds with purity higher than 80% were obtained. Preparative HPLC was used for further purification and three target compounds with purity higher than 98% were obtained. The compounds were identified as chlorogenic acid, pedaliin and pedaliin-6″-acetate.