Cations impact the biodegradation of iodosulfuron-methyl herbicidal ionic liquids by fungi.

In the framework of this study, six fungal isolates which demonstrated a high capability for biodegrading iodosulphuron-methyl sodium as well as herbicidal ionic liquids based on this herbicide were isolated from different soil samples. The isolates were identified based on the ITS region, whereas biodegradation residues were determined based on LC-MS/MS. Depending on the isolate, the half-lives values of the biodegraded herbicide or herbicidal ionic liquid ranged significantly from just 1.25 days to more than 40 days. The research findings unveiled that the structure of cations is a central limiting factor affecting fungal growth and herbicide transformation in case of ionic liquids. The length of the alkyl chain has been identified as the primary driver of herbicide toxicity, emphasizing the importance of structural factors in herbicide design. In cases when dodecyl(2-hydroxyethyl)dimethyl cation was used, its biodegradation ranged from 0 to approx. 20% and the biodegradability of the iodosulfuron-methyl was notably limited for the majority of the studied isolates. This knowledge provides guidance for development and selection of herbicides with reduced environmental impact. This study highlights the ecological importance of soil fungi, their potential role in herbicide biodegradation, the influence of cations on fungal growth and herbicide transformation, and the structural factors governing herbicide toxicity. Further research in these areas may lead to more efficient and environmentally friendly approaches to herbicide management.

Sorption of ionic liquids in soil enriched with polystyrene microplastic reveals independent behavior of cations and anions.

Recently, much attention has been focused on the application of the Ionic Liquids (ILs) with herbicidal activity in agriculture. It has been suggested that through the appropriate selection of cations and anions, one can adjust the properties of ILs, particularly the hydrophobicity, solubility, bioavailability, toxicity. In practical agricultural conditions, it will be beneficial to reduce the mobility of herbicidal anions, such as the commonly applied 2,4-dichlorophenoxyacetic acid [2,4-D] in the soil. Furthermore, microplastics are becoming increasingly prevalent in the soil, potentially stimulating herbicidal sorption. Therefore, we investigated whether cations in ILs influence the mobility of anions in OECD soil supplemented with polystyrene microplastic (PS). For this purpose, we used the 2,4-D based ILs consisting of: a hydrophilic choline cation [Chol][2,4-D] and a hydrophobic choline cation with a C12chain [C12Chol][2,4-D]. Characterization of selected micropolystyrene was carried out using the BET sorption-desorption isotherm, particle size distribution and changes in soil sorption parameters such as soil sorption capacity and cation exchange capacity. Based on the batch sorption experiment, the effect of microplastic on the sorption of individual cations and anions in soil contaminated with micropolystyrene was evaluated. The results obtained indicate that the introduction of a 1-10% (w/w) PS resulted in an 18-23% increase of the soil sorption capacity. However, the sorption of both ILs' cations increased only by 3-5%. No sorption of the [2,4-D] anion was noted. This suggests that cations and anions forming ILs, behave independently of each other in the environment. The results indicate the fact that ILs upon introduction into the environment are not a new type of emerging contaminant, but rather a typical mixture of ions. It is worth noting that when analyzing the behavior of ILs in the environment, it is necessary to follow the fate of both cations and anions.

Multi-faceted analysis of bacterial transformation of nitrofurantoin.

Excessive presence of antibiotics and their residues can be dangerous to the natural environment. To reduce this negative effect, efficient strategies to remove them from the ecosystem are required. This study aimed to explore the potential of bacterial strains to degrade nitrofurantoin (NFT). Single strains isolated from contaminated areas, namely Stenotrophomonas acidaminiphila N0B, Pseudomonas indoloxydans WB, and Serratia marcescens ODW152 were employed in this study. Degradation efficiency and dynamic changes within the cells during NFT biodegradation were investigated. For this purpose, atomic force microscopy, flow cytometry, zeta potential, and particle size distribution measurements were applied. Serratia marcescens ODW152 showed the highest performance in removal of NFT (96 % in 28 days). The AFM images revealed modifications of cell shape and surface structure induced by NFT. Zeta potential showed significant variations during biodegradation. Cultures exposed to NFT had a broader size distribution than the control cultures due to increased cells agglomeration or aggregation. 1-Aminohydantoin and semicarbazide were detected as nitrofurantoin biotransformation products. They showed increased cytotoxicity toward bacteria as determined by spectroscopy and flow cytometry. Results of this study suggest that nitrofurantoin biodegradation leads to formation of stable transformation products that significantly affect the physiology and structure of bacterial cells.

2,4-D versus 2,4-D based ionic liquids: Effect of cation on herbicide biodegradation, tfdA genes abundance and microbiome changes during soil bioaugmentation.

The commercial formulations of herbicides rely on surfactants which increase the efficiency of active substance. Herbicidal ionic liquids (ILs), in which cationic surfactants are combined with herbicidal anions, allow for additives' reduction and ensure very good herbicide performance with lower doses. We aimed to test the impact of synthetic and natural cations on biological degradation of 2,4-dichlorophenoxyacetic acid (2,4-D). Although primary biodegradation was high, the mineralization in agricultural soil indicated incomplete conversion of ILs to CO2. Even the introduction of naturally-derived cations resulted in an increase in the herbicide's half-lives - from 32 days for [Na][2,4-D] to 120 days for [Chol][2,4-D] and 300 days for the synthetic tetramethylammonium derivative [TMA][2,4-D]. Bioaugmentation with 2,4-D-degrading strains improves the herbicides' degradation, which was reflected by higher abundance of tfdA genes. Microbial community analysis confirmed that hydrophobic cationic surfactants, even those based on natural compounds, played a negative role on microbial biodiversity. Our study provides a valuable indication for further research related to the production of a new generation of environmentally friendly compounds. Moreover, the results shed a new light on the ionic liquids as independent mixtures of ions in the environment, as opposed to treating them as new type of environmental pollutants.

Application of phosphonium deep eutectic solvents as extractants in ultrasound-assisted dispersive liquid-liquid microextraction for preconcentration of trace amounts of herbicides in drainage ditches waters.

In this study, an efficient preconcentration method was presented that is based on dispersive liquid-liquid microextraction taking the advantage of newly synthesized phosphonium deep eutectic solvents used as extractants and ultrasound probe as a dispersing agent. The extracts obtained were analyzed by high-performance liquid chromatography. To optimize the five most important factors for the microextraction procedure a central composite design plan was used. Under optimal conditions (140 µl of extractant, 60 mg of NaCl, pH = 2.0, 120 s of extraction time with ultrasound probe as the dispersing agent, 16 min of centrifugation for phase separation), the proposed method allowed to achieve good precision with RSD between 3.2% and 9.7% at 1.0, 5.0 and 40.0 ng ml levels. The preconcentration factors were equal to 42, 39, and 41, and the limits of detection 0.128, 0.103, and 0.135 ng/ml for dicamba, 2-methyl-4-chlorophenoxyacetic acid, and 2-methyl-4-chlorophenoxypropionic acid, respectively. The proposed method was successfully applied for the determination of chlorophenoxy acid herbicides in water samples from drainage ditches with a good recovery in the range of 70%-93%.

Dynamics of microbial communities during biotransformation of nitrofurantoin.

The purpose of this research was to investigate the biodegradation of nitrofurantoin (NFT), a typical nitrofuran antibiotic of potential carcinogenic properties, by two microbial communities derived from distinct environmental niches - mountain stream (NW) and seaport water (SS). The collected environmental samples represent the reserve of the protected area with no human intervention and the contaminated area that concentrates intense human activities. The structure, composition, and diversity of the communities were analyzed at three timepoints during NFT biodegradation. Comamonadaceae (43.2%) and Pseudomonadaceae (19.6%) were the most abundant families in the initial NW sample. The top families in the initial SS sample included Aeromonadaceae (31.4%) and Vibrionaceae (25.3%). The proportion of the most abundant families in both consortia was remarkably reduced in all samples treated with NFT. The biodiversity significantly increased in both consortia treated with NFT suggesting that NFT significantly alters community structure in the aquatic systems. In this study, NFT removal efficiency and transformation products were also studied. The biodegradation rate decreased with the increasing initial NFT concentration. Biodegradation followed similar pathways for both consortia and led to the formation of transformation products: 1-aminohydantoin, semicarbazide (SEM), and hydrazine (HYD). SEM and HYD were detected for the first time as NFT biotransformation products. This study demonstrates that the structure of the microbial community may be directly correlated with the presence of NFT. Enchanced biodiversity of the microbial community does not have to be correlated with increase in functional capacity, such as the ability to biodegradation because higher biodiversity corresponded to lower biodegradation. Our findings provide new insights into the effect of NFT contamination on aquatic microbiomes. The study also increases our understanding of the environmental impact of nitrofuran residues and their biodegradation.

Variation in the Content of Bioactive Compounds in Infusions Prepared from Different Parts of Wild Polish Stinging Nettle (Urtica dioica L.).

Nettle is a common plant that offers many health benefits and is grown all over the world. The content of active compounds in roots, stems, and leaves was determined based on the extraction procedure optimized using the Central Composite Design. Flavonols, phenolic acids, trigonelline, nicotinamide, nicotinic acids, and short-chain organic acids were determined with the use of LC-MS/MS and capillary isotachophoresis. Trigonelline, which was not previously reported in the roots and stems of nettle, was found in all parts of the plant and considerable variations in its content were observed (2.8-108 µg g-1). Furthermore, the Principal Component Analysis taking into account more variables demonstrated differences in the content of bioactive components between roots and aerial parts of nettle.

A polydimethylsiloxane/deep eutectic solvent sol-gel thin film sorbent and its application to solid-phase microextraction of parabens.

A novel, efficient and reproducible thin-film solid-phase microextraction method using the polydimethylsiloxane/deep eutectic solvent sol-gel as coating material was developed for preconcentration of parabens from environmental samples. New sorbent materials consisting of polydimethylsiloxane with nine new synthesized deep eutectic solvents were prepared by the sol-gel method, and these materials were used to prepare thin film coatings on a mesh support. Among obtained compounds, the highest efficiency in the extraction of parabens was demonstrated by a sorbent consisting of trihexyltetradecylphosphonium chloride and n-docosanol in a molar ratio 1:2 mixed with polydimethylsiloxane. For the proposed method, the parameters affecting the extraction efficiency of parabens were optimized, including the coating material type based on polydimethylsiloxane/deep eutectic solvent, the sample volume, the salting-out effect, the pH of the sample, the extraction time, the desorption solvent and time. The limits of detection and quantitation were from 0.023 to 0.062 ng mL-1 and 0.076-0.201 ng mL-1, respectively, and the enrichment factors ranged from 174 to 186. Under optimal conditions, the proposed method based on measurements for river and lake water achieved good precision between 2.7 and 5.9% and recovery ranging from 79.1 to 88.1%. The developed method was successfully applied to determine trace levels of parabens in the samples of surface waters collected from rivers and lakes.

Development of novel thin-film solid-phase microextraction materials based on deep eutectic solvents for preconcentration of trace amounts of parabens in surface waters.

A green and sensitive thin-film solid-phase microextraction method based on deep eutectic solvent was developed that enables simultaneous isolation, preconcentration, and determination of parabens in surface waters. Six new deep eutectic solvents were synthesized and used directly to prepare thin-film coatings on a stainless steel mesh support. Among the compounds obtained, the highest efficiency in the extraction of parabens was found for a material consisting of trihexyltetradecylphosphonium chloride and n-docosanol in a molar ratio of 1:2. For the proposed method, parameters affecting the extraction efficiency of parabens, such as the coating material, the desorption solvent, the volume of the sample, the pH of the sample, the extraction and desorption time, and the salting-out effect, were optimized. Under optimal conditions, the proposed method allowed us to achieve good precision between 3.6 and 6.5% and recovery ranging from 68.1 to 91.4%. The limits of detection range from 0.018 to 0.055 ng/mL.

Azole fungicides: (Bio)degradation, transformation products and toxicity elucidation.

The increasing consumption of azole antifungal agents leads to their uncontrolled release into the environment. Therefore, it is crucial to remove their residues from natural ecosystems. This study aimed to examine the biological and chemical degradation of four typical azole fungicides: fluconazole (Fc), clotrimazole (Cl), climbazole (Cb), and epoxiconazole (Ep). The biodegradation was investigated using activated sludge and two novel Gram-negative bacterial strains. The chemical degradation experiments aimed to assess the efficiency of fungicides removal through UV treatment, the Fenton reaction, and a combination of these methods. Transformation products of Cb, Ep, and Cl photocatalytic removal were identified by mass spectrometry. In addition, the AlamarBlue® Assay and the MTT Assay allowed careful evaluation of the toxicity of azole derivatives and their transformation products towards newly isolated strains, Stenotrophomonas maltophilia AsPCl2.3 and Pseudomonas monteilii LB2. Among all azole fungicides, Cb was the most susceptible to biological removal while Fc, Ep, and Cl were basically resistant to biodegradation. Cl and Ep showed a significant biosorption on the activated sludge. Under optimized photolysis conditions, the removal efficiency of Cl, Cb, and Ep was significantly higher than that of biodegradation. The Fenton reaction supported by the UV-irradiation offered the best results of fungicides elimination. After 1 min of the experiment, Cl was almost completely removed while Cb and Ep removal rates reached an average of 60%. The proposed main degradation route of azole fungicides during UV-irradiation includes halogen atoms substitution by hydroxyl moieties. The final degradation product was imidazole or triazole. Azole fungicides and their transformation products differently affected the metabolic activity of Gram-negative bacteria. Cl and Cb intermediates showed lower toxicity than parent compounds. The findings help better understand the environmental impact of azole fungicides, their degradation, and toxicity. They also stress the need for reducing their uncontrolled release to the environment.

Occurrence and dietary risk of bisphenols and parabens in raw and processed cow's milk.

Raw cow's milk collected from farmers and processed cow's milk purchased from local grocery stores were analysed for the presence of six bisphenols (bisphenol A, bisphenol S, bisphenol F, bisphenol AF, bisphenol B, and bisphenol E) and five parabens (methylparaben, ethylparaben, propylparaben, butylparaben, and benzylparaben). The analytes were determined in their unconjugated form and (after enzymatic deconjugation) as the sum of conjugated and unconjugated compounds. The results show the presence of bisphenols mainly in the processed milk bought in stores while parabens were found in all samples of both raw and processed cow's milk. The average concentration of bisphenol A found in milk from cartons (0.87 ng mL-1) was greater than in milk from plastic bottles (0.35 ng mL-1). No such difference was found for parabens. Also, no considerable difference between the content of conjugated and total bisphenols and parabens was found except for ethylparaben. The determined compounds were always found below 2.0 ng mL-1 and calculations of the hazard quotients and the hazard index have shown that consumption of such milk is safe.

Comprehensive Analysis of Metabolites in Brews Prepared from Naturally and Technologically Treated Coffee Beans.

Coffee is one of the most popular beverages in the world. Therefore, this study analyzed 49 coffee samples of Arabica and Robusta species of different geographical origins and the treatment of beans including three degrees of roasting with the use of LC-MS/MS. This is the first study to present a comprehensive analysis of Kopi Luwak coffee brew metabolites in comparison to fully washed coffees and the drying post-harvest treatment of Arabica or Robusta coffee brews. Kopi Luwak showed higher levels of caffeine and theophylline in comparison to the analyzed washed and unwashed Arabica coffees, as well as a different proportion of caffeoylquinic isomers. There was no difference between Kopi Luwak and other Arabica coffees in terms of the concentration of vitamin B3, amines, and phenolic acids. This was confirmed in PCA. The steaming and roasting of beans as well as the addition of black beans influence the concentration of 4-CQA and the nicotinic, ferulic, and quinic acids content.

Synthesis of Selected Mixed Oxide Materials with Tailored Photocatalytic Activity in the Degradation of Tetracycline.

The elimination of antibiotics occurring in the natural environment has become a great challenge in recent years. Among other techniques, the photocatalytic degradation of this type of pollutant seems to be a promising approach. Thus, the search for new photoactive materials is currently of great importance. The present study concerns the sol-gel synthesis of mono, binary and ternary TiO2-based materials, which are used as active photocatalysts. The main goal was to evaluate how the addition of selected components-zirconium dioxide (ZrO2) and/or zinc oxide (ZnO)-during the synthesis of TiO2-based materials and the temperature of thermal treatment affect the materials' physicochemical and photocatalytic properties. The fabricated mixed oxide materials underwent detailed physicochemical analysis, utilizing scanning-electron microscopy (SEM), X-ray diffraction (XRD), diffuse reflectance spectroscopy (DRS), energy-dispersive X-ray spectroscopy (EDS), low-temperature N2 sorption (BET model), X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). The synthesized mixed oxide materials were used as photocatalysts in the heterogeneous photodegradation of tetracycline (TC). The physicochemical properties of the fabricated photocatalysts, including morphology, crystalline and textural structure, as well as the pH of the reaction system in the photocatalytic tests, were taken into account in determining their photo-oxidation activity. LC-MS/MS analysis was used to identify the possible degradation products of the selected antibiotic.

Biodegradation and photo-Fenton degradation of bisphenol A, bisphenol S and fluconazole in water.

Bisphenol A, bisphenol S, and fluconazole are ubiquitous environmental pollutants and their removal from water is of utmost importance. As the biodegradation of these compounds is usually not enough effective, often other degradation methods are required. The study presents the difference between biodegradation and photo-Fenton degradation with a much higher efficiency obtained in the latter process. Levels of biodegradation and chemical degradation were assessed based on high-performance liquid chromatography determination. Optimization of the photo-Fenton removal of bisphenol A, bisphenol S, and fluconazole resulted in about 100 % primary degradation of both bisphenols during 10-20 min and almost 90 % primary degradation of fluconazole within an hour. Degradation products formed in the process were identified using liquid chromatography with mass spectrometry and showed central scission of bisphenol S with the formation of phenol and sulfuric acid while for bisphenol A and fluconazole the oxidation resulted in much smaller structural changes.

Significance of the presence of antibiotics on the microbial consortium in wastewater - The case of nitrofurantoin and furazolidone.

Antibiotics in wastewater leads to migration of pollutants and disrupts natural processes of mineralization of organic matter. In order to understand the mechanism of this, research was undertaken on the influence of nitrofurantoin (NFT) and furazolidone (FZD), on the behaviour of a consortium of microorganisms present in a model wastewater in a bioreactor. Our study confirmed biodegradation of the antibiotics by the microbial consortium, with the degradation efficiency within 10 days of 65% for FZD, but only 20% for NFT. The kinetic study proved that the presence of analysed antibiotics had no adverse effect on the microbes, but the consortium behaviour differ significantly with the NFT reducing the consumption of organic carbon in wastewater and increasing the production of extracellular biopolymeric and volatile organic compounds, and the FZD reducing assimilation of other carbon sources to a less extent, at the expense of cellular focus on biodegradation of this antibiotic.

Nitrofurazone Removal from Water Enhanced by Coupling Photocatalysis and Biodegradation.

(1) Background: Environmental contamination with antibiotics is particularly serious because the usual methods used in wastewater treatment plants turn out to be insufficient or ineffective. An interesting idea is to support natural biodegradation processes with physicochemical methods as well as with bioaugmentation with efficient microbial degraders. Hence, the aim of our study is evaluation of the effectiveness of different methods of nitrofurazone (NFZ) degradation: photolysis and photodegradation in the presence of two photocatalysts, the commercial TiO2-P25 and a self-obtained Fe3O4@SiO2/TiO2 magnetic photocatalyst. (2) Methods: The chemical nature of the photocatalysis products was investigated using a spectrometric method, and then, they were subjected to biodegradation using the strain Achromobacter xylosoxidans NFZ2. Additionally, the effects of the photodegradation products on bacterial cell surface properties and membranes were studied. (3) Results: Photocatalysis with TiO2-P25 allowed reduction of NFZ by over 90%, demonstrating that this method is twice as effective as photolysis alone. Moreover, the bacterial strain used proved to be effective in the removal of NFZ, as well as its intermediates. (4) Conclusions: The results indicated that photocatalysis alone or coupled with biodegradation with the strain A. xylosoxidans NFZ2 leads to efficient degradation and almost complete mineralization of NFZ.

Application of the electropolymerized poly(3,4-ethylenedioxythiophene) sorbent for solid-phase microextraction of bisphenols.

A new, simple, and effective procedure using poly(3,4-ethylenedioxythiophene)/lignosulfonate electropolymerized sorbent solid-phase microextraction (PEDOT/LS-SPME) combined with LC-MS/MS for determination of bisphenols in environmental water samples was developed. Various parameters influencing the performance of the analytical procedure including the type of sorbent, electropolymerization time, sorbent preconditioning time, extraction time, desorption (time and solvent), and sample pH were investigated and optimized. Under optimal conditions the proposed method allowed us to achieve good precision (n = 5) between 6.0 and 12.1%. The limits of detection were equal to 0.17 µg L-1 for BPA, 0.16 µg L-1 for BPF, 0.07 µg L-1 for BPE, 0.05 µg L-1 for BPB, and 0.027 µg L-1 for BPAF. The proposed method was successfully applied for the determination of bisphenols in aqueous environmental samples.

The presence of bisphenol A in the thermal paper in the face of changing European regulations - A comparative global research.

Bisphenol A (BPA) is used as a color developer in a thermal paper that after a heating process reacts with a leuco dye and changes it to a colored form. Receipts from cash registers are considered as the main source of consumer exposure to bisphenols together with polycarbonates and epoxy resins. Levels of BPA and its possible alternatives were determined in thermal paper samples collected between May 2018 and May 2019 in 22 European and 17 non-European countries on all inhabited continents (220 samples in total, 133 of which were from Europe and 87 from other countries). These measurements were intended to check the level of BPA presence in receipts originating from different countries, especially from Europe in the light of changing regulations restricting its use. The effect of thermal printing on developer content was also analyzed, but no major changes in concentrations of bisphenols were observed during the process. Thus, printed receipts could be used for the determination of bisphenol content. Analysis of receipts from 39 countries has shown that BPA is still the most common compound used around the world with 69% samples containing this color developer. Among other tested bisphenols, BPS was used as a color developer in 20% samples, but it was noted that all samples collected from Japan and the United States of America were found to contain only BPS. Other bisphenols (F, AF, E, and B) considered as possible alternatives for BPA were detected only at trace levels or not detected at all, which showed that they were not used as color developers. The relatively large use of BPS as a BPA substitute is worrying because this compound not only has similar endocrine properties but is also poorly biodegradable. Besides, its relatively high polarity facilitates spreading in the environment.

Removal of Bisphenol A and Its Potential Substitutes by Biodegradation.

The possibility of removing bisphenol A and its five potential substitutes (bisphenols S, F, AF, E, and B) was tested using microorganism consortia from river water and activated sludge from municipal and rural wastewater treatment plants. For most bisphenols, biodegradation with activated sludge was faster than with river water and a greater extent of biodegradation was also achieved. However, only bisphenol A and bisphenol F underwent 100% primary biodegradation while other bisphenols degraded no more than about 50% which has some important implications in case of their increased usage. Metabolic activity in biodegradation liquors was also tested and it showed higher activity in the tests with activated sludge than with river water. However, there was no clear connection between the decline of metabolic activity and the extent of biodegradation as decreased activity was observed for two easily degrading bisphenols and two others with little biodegradability. It can be assumed that two different phenomena are involved in this process including depletion of nutrients for easily degradable bisphenol A and absence of nutrients for bacteria incapable of primary degradation of bisphenol AF and bisphenol S.

Robust biodegradation of naproxen and diclofenac by laccase immobilized using electrospun nanofibers with enhanced stability and reusability.

Enzymatic biodegradation of pharmaceuticals, using enzymes such as laccase, is a green solution for the removal of toxic pollutants that has attracted growing interest over recent years. Moreover, the application of immobilized biocatalysts is relevant for industrial applications, due to the improved stability and reusability of the immobilized enzymes. Thus, in the present study, laccase was immobilized by adsorption and encapsulation using poly(l-lactic acid)-co-poly(ε-caprolactone) (PLCL) electrospun nanofibers as a tailor-made support. The produced biocatalytic systems were applied in the biodegradation of two commonly used anti-inflammatories, naproxen and diclofenac, which are present in wastewaters at environmentally relevant concentrations. The results showed that under optimal process conditions (temperature 25 °C, pH 5 and 3 for naproxen and diclofenac respectively), even from a solution at a concentration of 1 mg L-1, over 90% of both pharmaceuticals was removed by encapsulated laccase in batch mode. Both immobilized enzymes also exhibited high reusability: after five reaction cycles approximately 60% and 40% of naproxen and diclofenac were removed by encapsulated and adsorbed laccase respectively. In addition, a thorough analysis was made of the products of biodegradation of the two studied pollutants. Furthermore, toxicity study of the mixture after biodegradation of the pharmaceuticals showed that the solutions obtained after the process were approximately 65% less toxic than the initial naproxen and diclofenac solutions.