Limitations and Challenges in the Stability of Cysteamine Eye Drop Compounded Formulations

Accumulation of cystine crystals in the cornea of patients suffering from cystinosis is considered pathognomonic and can lead to severe ocular complications. Cysteamine eye drop compounded formulations, commonly prepared by hospital pharmacy services, are meant to diminish the build-up of corneal cystine crystals. The objective of this work was to analyze whether the shelf life proposed for six formulations prepared following different protocols used in hospital pharmacies is adequate to guarantee the quality and efficacy of cysteamine eye drops. The long-term and in-use stabilities of these preparations were studied using different parameters: content of cysteamine and its main degradation product cystamine; appearance, color and odor; pH and viscosity; and microbiological analysis. The results obtained show that degradation of cysteamine was between 20% and 50% after one month of storage in the long-term stability study and between 35% and 60% in the in-use study. These data confirm that cysteamine is a very unstable molecule in aqueous solution, the presence of oxygen being the main degradation factor. Saturation with nitrogen gas of the solutions offers a means of reducing cysteamine degradation. Overall, all the formulae studied presented high instability at the end of their shelf life, suggesting that their clinical efficacy might be dramatically compromised.

The ChpR transcriptional regulator of Sinorhizobium meliloti senses 3,5,6-trichloropyridinol, a degradation product of the organophosphate pesticide chlorpyrifos

The global use of organophosphate insecticides (OPPs) and the growing concern of off-target side effects due to OPP exposure has prompted the need for sensitive and economical detection methods. Here we set out to engineer a previously identified OPP responsive transcription factor, ChpR, from Sinorhizobium melilotii to respond to alternative OPPs and generate a repertoire of whole-cell biosensors for OPPs. The ChpR transcription factor and cognate promoter P chpA, have been shown to activate transcription in the presence of the OPP chlorpyrifos (CPF). Utilizing a GFP reporter regulated by ChpR in a whole-cell biosensor we found that the system responds significantly better to 3,5,6-trichloro-2-pyridinol (TCP), the main degradation product of CPF, compared to CPF itself. This biosensor was able to respond to TCP at 390 nM within 4 h compared to 50 µM of CPF in 7 h. The ChpR-P chpA , and the activating ligand TCP, were able to regulate expression of a kanamycin resistance/sucrose sensitivity (kan/sacB) selection/counterselection module suitable for high throughput mutagenesis screening studies. The ability to control both GFP and the kan/sacB module demonstrates the utility of this reporter for the detection of CPF affected areas. The ChpR-P chpA system serves as an additional positive regulator switch to add to the growing repertoire of controllers available within synthetic biology.

The Role of Non-Enzymatic Degradation of Meropenem—Insights from the Bottle to the Body

Several studies have addressed the poor stability of meropenem in aqueous solutions, though not considering the main degradation product, the open-ring metabolite (ORM) form. In the present work, we elucidate the metabolic fate of meropenem and ORM from continuous infusion to the human bloodstream. We performed in vitro infusate stability tests at ambient temperature with 2% meropenem reconstituted in 0.9% normal saline, and body temperature warmed buffered human serum with 2, 10, and 50 mg/L meropenem, covering the therapeutic range. We also examined meropenem and ORM levels over several days in six critically ill patients receiving continuous infusions. Meropenem exhibited a constant degradation rate of 0.006/h and 0.025/h in normal saline at 22 °C and serum at 37 °C, respectively. Given that 2% meropenem remains stable for 17.5 h in normal saline (≥90% of the initial concentration), we recommend replacement of the infusate every 12 h. Our patients showed inter-individually highly variable, but intra-individually constant molar ORM/(meropenem + ORM) ratios of 0.21–0.52. Applying a population pharmacokinetic approach using the degradation rate in serum, spontaneous degradation accounted for only 6% of the total clearance.

Stability Analyses of Guanitoxin Isolated from Sphaerospermopsis torques-reginae by Mass Spectrometry

Guanitoxin (GNT) is a natural organophosphate produced by some species of freshwater cyanobacteria, which inhibits the active site of acetylcholinesterase, preventing the hydrolysis of cholinesterases and consequently causing serious disturbances in the neuromuscular system. Despite having a chemical structure like synthetic organophosphates, there is still no analytical standard available for environmental and freshwater monitoring. Therefore, this study investigated the stability of GNT under different storage conditions, pH, and temperature. The toxin is produced by the cyanobacterium Sphaerospermopsis torques-reginae and monitored by liquid chromatography coupled to mass spectrometry (LC-MS) and LC-MS/MS for the identification and verification of its stability. The main degradation product formed is the hydroxy-amino-guanidinic derivative of the toxin. The results also indicate that GNT is stable in acidic medium (pH = 3.0), but can gradually degrade at room temperature (> 23 ºC) over a period of 96 h. Lyophilized biomass of S. torques-reginae containing GNT remained stable when stored in a refrigerator below 4 ºC. In addition, the extraction yield is higher when prepared from fresh S. torques-reginae cells than from lyophilized material. Thus, the results shown here contribute with valuable information for studies that aim at the isolation, identification, and monitoring of GNT in samples of raw water and cyanobacterial blooms.

Determination of Mancozeb, a Pesticide Used Worldwide in Agriculture: Comparison among GC, LC, and CE

Background: The determination of mancozeb, a fungicide extensively used in agriculture, is a challenge, due to the nature of the compound, a manganese and zinc complex of ethylenebis dithiocarbamate and because of the general instability of the dithiocarbamates. Methods: Mancozeb was analyzed in a GC-EI-MS system after derivatization by CE-UV with detection at 280 nm and in LC-ESI-MS-MS in MRM mode. Results: A comparative study of the performance of three different techniques for the detection of mancozeb was explored, highlighting the advantages and drawbacks of them. The limits of detection and quantification of the techniques were determined; the repeatability was assessed, showing values of relative standard deviation. Gas chromatography, although very sensitive, was not reproducible enough due to fast degradation of the derivatization product, whereas capillary electrophoresis-UV showed problems in run-to-run reproducibility which had the worst limit of detection. LC coupled with tandem mass spectrometry was the most reliable and precise technique and was able to determine the main degradation product of Mancozeb, at the same time. The proposed LC procedure was verified by applying it to a commercial formulation, a fungicide of known concentration, and to Italian white grapes treated with the formulation sprayed during cultivation. Conclusion: Thanks to the simplified sample handling, the proposed method resulted to be simple, fast, green, economic, and suitable for residue analysis in grapes and other fruits. Finally, the method was compared with other similar investigations.

Effects of Glyphosate-Based and Derived Products on Sea Urchin Larval Development

Despite the widespread use of herbicide glyphosate in cultivation, its extensive runoff into rivers and to coastal areas, and the persistence of this chemical and its main degradation product (aminomethylphosphonic acid, AMPA) in the environment, there is still little information on the potential negative effects of glyphosate, its commercial formulation Roundup® and AMPA on marine species. This study was conducted with the aim of providing a comparative evaluation of the effects of glyphosate-based and its derived chemicals on the larval development of the sea urchin Paracentrotus lividus, thus providing new data to describe the potential ecotoxicity of these contaminants. In particular, the effects on larval development, growth and metabolism were assessed during 48 h of exposure from the time of egg fertilization. The results confirm that AMPA and its parent compound, glyphosate have similar toxicity, as observed in other marine invertebrates. However, interestingly, the Roundup® formulation seemed to be less toxic than the glyphosate alone.

The Photocatalytic Degradation of 2,3,7,8-Tetrachlorodibenzo-p-Dioxin in the Presence of Silver–Titanium Based Catalysts

Polychlorinated dibenzo-p-dioxins (PCDD) are persistent toxic compounds that are ubiquitous in the environment. The photodegradation of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in the presence of silver titanium oxide (AgTi) and silver titanium doped into the Y-zeolite (AgTiY) was tested using high (254 nm) and mid (302 nm) energy UV irradiation sources. AgTi and AgTiY, both showed success in the photodegradation of 2,3,7,8-TCDD dissolved in methanol/tetrahydrofuran solution. Both catalysts were found to effectively decompose TCDD at 302 nm (lower energy) reaching in between 98–99% degradation after five hours, but AgTiY showed better performance than AgTi at 60 min reaching 91% removal. Byproducts of degradation were evaluated using Gas chromatography/mass spectrometry (GC–MS), resulting in 2,3,7-trichlorodibenzo-p-dioxin, a lower chlorinated congener and less toxic, as the main degradation product. Enzyme Linked Immunosorbent Assay (ELISA) was used to evaluate the relative toxicity of the degradation byproducts were a decrease in optical density indicated that some products of degradation could be potentially more toxic than the parent TCDD. On the other hand, a decrease in toxicity was observed for the samples with the highest 2,3,7,8-TCDD degradation, confirming that AgTiY irradiated at 302 nm is an excellent choice for degrading TCDD. This is the first study to report on the efficiency of silver titanium doped zeolites for the removal of toxic organic contaminants such as dioxins and furans from aquatic ecosystems.

pH effect on stability and kinetics degradation of nitazoxanide in solution

Stability studies correspond to a set of tests designed to assess changes in the quality of a given drug over time and under the influence of a number of factors. Among these factors, pH plays an important role, due to the catalytic effect that hydronium and hydroxide ions can play in several reactions. In the present study, the degradation kinetics of nitazoxanide was evaluated over a wide pH range, and the main degradation product generated was identified by LC-MS/MS. Nitazoxanide showed first-order degradation kinetics in the pH range of 0.01 to 10.0 showing greater stability between pH 1.0 and 4.0. The degradation rate constant calculated for these pH was 0.0885 x 10-2 min-1 and 0.0689 x 10-2 min-1, respectively. The highest degradation rate constant value was observed at pH 10.0 (0.7418 x 10-2 min-1) followed by pH 0.01 (0.5882 x 10-2 min-1). A major degradation product (DP-1) was observed in all conditions tested. Through LC-MS/MS analysis, DP-1 was identified as a product of nitazoxanide deacetylation. The effect of pH on the stability of nitazoxanide and the kinetic data obtained contribute to a better understanding of the intrinsic stability characteristics of nitazoxanide.

Leaching and degradation of 13C2-15N-glyphosate in field lysimeters

Glyphosate (GLYP), the globally most important herbicide, may have effects in various compartments of the environment such as soil and water. Although laboratory studies showed fast microbial degradation and a low leaching potential, it is often detected in various environmental compartments, but pathways are unknown. Therefore, the objective was to study GLYP leaching and transformations in a lysimeter field experiment over a study period of one hydrological year using non-radioactive 13C2-15N-GLYP labelling and maize cultivation. 15N and 13C were selectively measured using isotopic ratio mass spectrometry (IR-MS) in leachates, soil, and plant material. Additionally, HPLC coupled to tandem mass spectrometry (HPLC-MS/MS) was used for quantitation of GLYP and its main degradation product aminomethylphosphonic acid (AMPA) in different environmental compartments (leachates and soil). Results show low recoveries for GLYP (< 3%) and AMPA (< level of detection) in soil after the study period, whereas recoveries of 15N (11–19%) and 13C (23–54%) were higher. Time independent enrichment of 15N and 13C and the absence of GLYP and AMPA in leachates indicated further degradation. 15N was enriched in all compartments of maize plants (roots, shoots, and cobs). 13C was only enriched in roots. Results confirmed rapid degradation to further degradation products, e.g., 15NH4+, which plausibly was taken up as nutrient by plants. Due to the discrepancy of low GLYP and AMPA concentrations in soil, but higher values for 15N and 13C after the study period, it cannot be excluded that non-extractable residues of GLYP remained and accumulated in soil.