A simple and rapid spectrophotometric method for the determination of trans-chalcone in raw-materialand topical formulation / Método espectrofotométrico simples e rápido paradeterminação de trans-chalcona em matéria-primae em formulação tópica

Trans-chalcone (TC) is a flavonoid precursor characterized by a wide spectrum of action, with anti-inflammatory and antioxidant effects. However, no validated methods are available in official compendia for the analysis of this substance. Thus, the aim of this work was to develop and validate a simple, fast, and reproducible spectrophotometric method for the analysis of TC in raw material, and in topical pharmaceutical formulation containing TC. The established conditions were: methanol as extracting solvent, and detection wavelength of 309 nm by UV spectrophotometer. All tests followed the rules of Resolution RDC 166, 2017. The proposed method was selective. Linearity was demonstrated in the concentration range of 1 to 8 µg/mL (r = 0.999). Repeatability and intermediate precision were confirmed by low relative standard deviation values of 1.53% and 2.70% for TC, and of 1.73% and 2.91% for formulation containing TC. Accuracy, evaluated through recovery test, was adequate, with minimum of 98.24% and maximum of 100.23% of recovery. It was observed that the small deliberate modifications done did not interfere with the results, demonstrating the method is robust. The results showed that the method was considered suitable for the intended purpose, inexpensive, easy to apply, selective, linear, precise, accurate, and robust for the determination TC, and pharmaceutical formulation containing TC. Thus, the method developed satisfies the need for an analytical method for the determination of TC, and topical formulation containing TC, being effective, innovative and able to aid in the development of the pharmaceutical field.

Trans-chalcona (TC) é um precursor de flavonoides caracterizado por um amplo espectro de ação, como efeitos anti-inflamatórios e antioxidantes. No entanto, não há método validado disponível em compêndio oficial para análise deste composto. Então, o objetivo deste trabalho foi desenvolver e validar um método espectrofotométrico, simples, rápido e reprodutível para análise de TC em matéria-prima, e em formulação farmacêutica tópica contendo TC. As condições estabelecidas foram: metanol como o solvente de extração, e detecção no comprimento de onda de 309 nm por espectrofotometria no UV. Todos os testes seguiram as normas da RDC 166, 2017. O método proposto foi seletivo. A linearidade foi demonstrada na faixa de concentração de 1 a 8 µg/mL (r = 0.999). A repetibilidade e a precisão intermediária foram confirmadas pelos valores baixos de desvio padrão relativo de 1,53% e 2,70% para a TC, e de 1,73% e 2,91% para a formulação contendo TC. A exatidão, avaliada por meio de testes de recuperação, foi adequada, com mínimo de 98,24% e máximo de 100,04% de recuperação. Observou-se que pequenas modificações no método não interferiram nos resultados, demonstrando que o método é robusto. Os resultados demonstraram que o método foi adequado para a finalidade pretendida, barato, de fácil aplicação, seletivo, linear, preciso, exato e robusto para determinação de TC, e de formulação contendo TC. Então o método desenvolvido satisfaz as necessidades de um método analítico para determinação de TC, e de formulação tópica contendo TC, e é eficaz, inovador e pode contribuir para o desenvolvimento da área farmacêutica.

Intraoperative pneumatic tourniquet application reduces soft-tissue microcirculation, but without affecting wound healing in calcaneal fractures.

BACKGROUND: Wound healing complications are a major challenge following the extended lateral approach in calcaneal fractures. Soft-tissue microcirculation plays an important role via the delivery of oxygen, nutrients, and the regulation of a local milieu. The aim of this clinical study was to examine the effect of intraoperative pneumatic tourniquet application on skin and subcutaneous microcirculation, and its impact on wound healing progression. METHODS: Patients with calcaneal fractures were randomly assigned to two groups defined by a surgery conducted either with use or without use of a tourniquet. Blood flow (BF [AU]), tissue oxygen saturation (SO2[%]) and the relative amount of haemoglobin (rHb[AU]) were intraoperatively measured at two depths (2 and 8 mm) non-invasively by spectrophotometry (Micro-Lightguide O2C®, LEA Medizintechnik, Giessen, Germany). Time points were before and after inflation of the pneumatic tourniquet and also at the end of surgery before deflation. A linear mixed model (LMM) was fitted for statistical analysis. RESULTS: Thirty-four patients (3 women and 31 men) with 37 calcaneal fractures were included. In 22 of them, the surgery was conducted with a tourniquet and in the other 15 without its use. A significant decrease of microcirculation, characterized by decreases in blood flow (p = 0.011) and tissue oxygenation (p = 0.023) was measured in 8 mm depth after inflating the tourniquet. However, these changes did not influence the time of postoperative wound healing. CONCLUSION: The use of a pneumatic tourniquet reduces deep microcirculation without affecting postoperative wound healing. Trial registration The study was registered in www. CLINICALTRIALS: gov (NCT01264146).

Selective microextraction of erythrosine (E127) in foodstuffs using a new generation high-density type-V deep eutectic solvent.

A novel and selective (deep eutectic solvent) DES-based microextraction method was established for the first time, utilizing a synthesized new generation High-Density Type-V DES for monitoring the dye Erythrosine (E127) in various foodstuffs and drugs. Type-V DES was created from acetophenone and diphenylamine at 3:1 M ratio. The pH, DES amount, and vortex time were optimized using Box-Behnken Design (BBD) of Response Surface Methodology (RSM). The quadratic microextraction model with R2 = 0.9982 was obtained. The limit of detection, preconcentration factor and linear dynamic range were determined to be 12 µg/L, 50 and 41-4000 µg/L, respectively. Effects of matrix components were examined. The developed High-Density Type-V Deep Eutectic Solvent Microextraction (HD-V-DES-ME) method was applied to foodstuffs and drugs to monitor their E127 contents and subsequently validated by applying spiked tests to real samples, with recoveries ranging between 94 and 101 %. The indexes of environmental friendliness and practicality for the method were evaluated using the Analytical GREEnness metric approach tool (AGREE) and the Blue Applicability Grade Index tool (BAGI), respectively.

Hyperspectral Imaging Database of Human Facial Skin.

The perceived color of human skin is the result of the interaction of environmental lighting with the skin. Only by resorting to human skin spectral reflectance, it is possible to obtain physical outcomes of this interaction. The purpose of this work was to provide a cured and validated database of hyperspectral images of human faces, useful for several applications, such as psychophysics-based research, object recognition, and material modeling. The hyperspectral imaging data from 29 human faces with different skin tones and sexes, under constant lighting and controlled movements, were described and characterized. Each hyperspectral image, which comprised spectral reflectance of the whole face from 400 to 720 nm in 10 nm steps at each pixel, was analyzed between and within nine facial positions located at different areas of the face. Simultaneously, spectral measurements at the same nine facial positions using conventional local point and/or contact devices were used to ascertain the data. It was found that the spectral reflectance profile changed between skin tones, subjects, and facial locations. Important local variations of the spectral reflectance profile showed that extra care is needed when considering average values from conventional devices at the same area of measurement.

Solvent Dependence of Ionic Liquid-Based Pt Nanoparticle Synthesis: Machine Learning-Aided In-Line Monitoring in a Flow Reactor.

Colloidal platinum nanoparticles (Pt NPs) possess a myriad of technologically relevant applications. A potentially sustainable route to synthesize Pt NPs is via polyol reduction in ionic liquid (IL) solvents; however, the development of this synthetic method is limited by the fact that reaction kinetics have not been investigated. In-line analysis in a flow reactor is an appealing approach to obtain such kinetic data; unfortunately, the optical featurelessness of Pt NPs in the visible spectrum complicates the direct analysis of flow chemistry products via ultraviolet-visible (UV-vis) spectrophotometry. Here, we report a machine learning (ML)-based approach to analyze in-line UV-vis spectrophotometric data to determine Pt NP product concentrations. Using a benchtop flow reactor with ML-interpreted in-line analysis, we were able to investigate NP yield as a function of residence time for two IL solvents: 1-butyl-1-methylpyrrolidinium triflate (BMPYRR-OTf) and 1-butyl-2-methylpyridinium triflate (BMPY-OTf). While these solvents are structurally similar, the polyol reduction shows radically different yields of Pt NPs depending on which solvent is used. The approach presented here will help develop an understanding of how the subtle differences in the molecular structures of these solvents lead to distinct reaction behavior. The accuracy of the ML prediction was validated by particle size analysis and the error was found to be as low as 4%. This approach is generalizable and has the potential to provide information on various reaction outcomes stemming from solvent effects, for example, differential yields, orders of reaction, rate coefficients, NP sizes, etc.

Efficient Absorbance-Based Assay for Rapid Antibiotic Susceptibility Testing of Enterobacterales.

It is increasingly important to rapidly receive information on the antimicrobial susceptibility of bacteria due to the rise in antimicrobial resistance worldwide. However, traditional phenotypic methods are time-consuming. Thus, the objective of this study was to develop a rapid assay that can detect antibiotic-resistant bacterial isolates phenotypically in less than 2 h. The microplate assay used in this study is based on absorbance measurements of pure bacterial isolates grown in liquid media with and without antibiotics. Absorbance was measured at the beginning of the assay and after 90 min of incubation at 37 °C. Susceptibility was calculated for bacterial isolates that, in the absence of antibiotics, exhibited more than a 50% growth increase by comparing the absorbance value of the culture in the presence of an antibiotic at 90 min with its initial value. Here, we show that it is possible to phenotypically screen the antibiotic susceptibility of Enterobacterales and Acinetobacter spp. isolates to three different antibiotics in 90 min. The test demonstrated an accuracy of 98.8%, sensitivity of 97.6%, and specificity of 99.6%. The false susceptibility rates were 0.2% and false resistance rates were 1.0%. This rapid and simple absorbance test has proven suitable for the screening of antibiotic susceptibility for a large number of strains with high accuracy and sensitivity. This method can be performed without specialized and costly materials and/or equipment, which makes it highly suitable for laboratories with limited resources.

Assessing the Aesthetic Performance of CAD/CAM Provisional Restorative Materials: A Spectrophotometric Analysis.

Achieving color match between natural teeth and restorative materials is crucial in dentistry. Factors such as the light source, brightness, and opacity influence tooth color, determined by light absorption and scattering within the material. Advances in CAD/CAM systems have enhanced prosthodontic treatments, particularly with new temporary materials, but data on their color stability and masking ability remains limited. However, data on the color stability and masking ability of these CAD/CAM materials is limited. Telio® CAD-Temp and VITA CAD-Temp® blocks were cut into slices and polished. Composite resin specimens were prepared using a custom-designed metal resin former and light-cured. Samples were paired randomly and assigned to experimental groups based on base type and thickness (n = 30). Samples were stored in a controlled environment for 24 h before color evaluation using an EasyShade® V spectrophotometer. Color difference (ΔE) was calculated using L*, a*, and b* values. Statistical analyses included the Shapiro-Wilk test, Levene's test, and three-way ANOVA, with post-hoc comparisons using the Bonferroni method (α = 0.05). ΔE was classified according to perceptibility (PT = 1.2) and acceptability (AT = 2.7) values.

Intelligent spectrophotometric resolution platforms for the challenging spectra of ipratropium and fenoterol in their combination inhaler with ecological friendliness assessment.

Asthma and chronic obstructive pulmonary disease (COPD) are the most common diagnoses for adults and children with respiratory tract inflammation. Recently, a novel fixed dose combination consisting of Ipratropium and Fenoterol has been released for the management and control of the symptoms of such disorders. The current research has newly developed and optimized three smart, accurate, simple, cost-effective, and eco-friendly spectrophotometric methods that enabled the simultaneous determination of the drugs under study in their combined inhaler dosage form, without the need for any previous separation steps, using water as a green solvent. The strategy employed was based on calculating one or two factors as a numerical spectrum or constant, which provided the complete removal of any component in the mixture that might overlap and the mathematical filtration of the targeted analyte. The methods developed could be classified into two types of spectrophotometric windows. Window I; involved absorption spectrum in their original zero-order forms (°D), which included recently designed methods named induced concentration subtraction (ICS) and induced dual wavelength (IDW). While window III focused on the ratio spectrum as the induced amplitude modulation (IAM) method. The extremely low absorptivity and lack of distinct absorption maximum in the zero-order absorption spectrum of Ipratropium were two intrinsic challenges that were better overcome by the proposed spectrophotometric methods than by the conventionally used ones. According to ICH guidelines, the proposed methods were validated using unified regression over range 2.0-40.0 µg/mL in the ICS method, while the linearity ranges for the IDW and IAM methods were 5.0-40.0 µg/mL of Ipratropium and 2.0-40.0 µg/mL of Fenoterol. Moreover, the three proposed methods were effectively used to assay the co-formulated marketed inhaler and further expanded to confirm the delivered dose uniformity in compliance with the USP guidelines. Finally, the established methods were evaluated for their greenness and blueness, in comparison to the official and reported analysis methods, using advanced cutting edge software metrics. Furthermore, the suggested techniques adhered well to the white analytical chemistry postulates that were recently published.

Magnetic solid-phase extraction of amoxicillin and doxycycline from water and urine samples based on cetylpyridinium chloride-modified Fe3O4 nanoparticles followed by spectrophotometric determination.

This research describes an easy, rapid, and inexpensive magnetic solid-phase extraction (MSPE) approach employing Fe3O4 magnetic nanoparticles modified with cetylpyridinium chloride (Fe3O4@CPC/MNPs) for extracting amoxicillin (AMX) and doxycycline (DOX) after derivatization with 4-chloroaniline as a color reagent. The azo-coupling of AMX and DOX with the color reagent in the alkaline medium caused yellow and yellow-orange azo dyes with maximum absorption wavelengths of 435 and 438 nm, respectively. The UV-Vis spectroscopy was utilized to determine the target analyte after the extraction procedure. Good linearities (R2 > 0.99) in the concentration ranges of 0.03-4.50 and 0.05-6.00 µg/mL were obtained for AMX and DOX, respectively. The experimental detection limits of AMX and DOX were obtained as 0.01 and 0.02 µg/mL, respectively. The developed approach was effectively applied to pre-concentrate and quantify AMX and DOX in environmental water and urine samples.

Evaluation of Spontaneous Overtime Methemoglobin Formation in Post-Mortem Blood Samples from Real Cases in Critical Storage Conditions.

Nitrite/nitrate poisoning is an emerging problem, with an ongoing escalation of reported self-administration with suicidal intent in several countries. Nitrites toxicity mainly consists of their interaction with hemoglobin (Hb), causing its oxidization to methemoglobin (MetHb). In order to give support to the correct procedures for the analysis of these cases, this study aims to evaluate spontaneous sample degradation and consequent MetHb formation in the typical storage conditions of a forensic toxicology laboratory. Two different types of samples have been used in this study: the first stage of our study consisted of a retrospective analysis of blood samples obtained by judicial autopsies already stored in the toxicology laboratory, collected over four years (2018-2021), while the samples used for the second stage were appositely collected during judicial autopsies. The data obtained by the application of a derivative spectrophotometry method on these samples suggest that there seems not to be a maximum threshold for MetHb formation within which it is possible to state with a sufficient grade of certainty that the concentration of MetHb found is consistent with an ante-mortem formation and is not the result of an artifact due to sample degradation and storage conditions. On the other hand, the results suggest that MetHb formation depends on the time passed between sample collection and analysis, so that a tempestive sample processing, performed as soon as the samples are received in the laboratory, is crucial to obtain the maximum reliability and diagnostic values from the data when MetHb quantitation is necessary.

Effect of surface sealant on the color stability and whiteness index of single-shade resin composites after staining and bleaching.

Objectives: The aim of the current study was to evaluate the effect of polishing systems and surface sealant on the color stability and whiteness index of single-shade resin composites after staining and bleaching. Materials and Methods: Three single-shade (Omnichroma, Charisma Diamond One, Zenchroma) and one multi-shade (Filtek Z250) materials were tested. From each resin composite, 40 specimens were prepared. The specimens were divided into 4 subgroups (n = 10) according to the surface treatments: 1-step polishing, 1-step + Biscover LV, 2-step polishing, and 2-step polishing + Biscover LV. Color differences (ΔE00) were calculated after being immersed in the coffee solution for 12 days. After the staining, the specimens were immersed in a whitening mouthrinse (Crest-3D White) for 12 hours. Whiteness index differences (∆WID = WID after staining - WID after bleaching) values were recorded. The generalized linear model was used for analysis (p < 0.05). Results: The lowest and highest ΔE00 values were found for Zenchroma and Charisma Diamond One respectively. Sealed groups indicated higher ΔE00 values than nonsealed groups with significant differences (p = 0.008). The lowest and highest ΔWID values were found for Zenchroma and Charisma Diamond One respectively. Sealed groups indicated lower ΔWID values than nonsealed groups with significant differences (p = 0.022). Conclusions: The use of surface sealant increased the discoloration and showed less whiteness change in resin materials. When the 1-step was compared with the 2-step polishing, the effects on the color stability and whiteness index values of the resin materials were similar.

Application of validated UV-Vis spectrophotometry-colorimetric methods for specific quantification of deferiprone in the development of iron-responsive nanoparticle loaded into dissolving microneedle.

Deferiprone (DFP) is one of the iron-chelating agents used in iron overload therapy for patients with ß-thalassemia major (ß-TM). However, the use of DFP is limited as it experiences a first-pass effect and can potentially cause iron deficiency due to uncontrolled release. Therefore, iron-responsive (NP-IR) DFP nanoparticle innovation was developed to control DFP release. A dissolving microneedle system (NP-IR-DMNs) was used to maximize DFP release. However, in support of this development, validation of analytical methods using spectrophotometry and colorimetrics was carried out. UV-Vis spectrophotometry is an approach that is easy to use, practical, and more cost-effective than others. The DFP levels were determined in normal and iron-overloaded medium solutions with 1%, 2%, and 4% concentrations. In addition, DFP levels were also measured in rat plasma using the colorimetric method with the addition of FeCl3 reagent to increase sensitivity for the detection of the analyte. The procedures used as guidelines in the validation procedure are The International Council for Harmonization (ICH). As a result, all linear correlation values of medium and plasma ≥ 0.999 were obtained. The LOQ levels obtained were 0.55 µg/mL, 0.44 µg/mL, 0.42 µg/mL, 0.52 µg/mL, and 1.01 µg/mL in plasma, 1% FeSO4, 2% FeSO4, 4% FeSO4, and normal media, respectively. The accuracy and precision were confirmed valid, as all values were within the requirements and did not change during dilution. Then, this approach was successfully applied to determine the levels of DFP in NP-IR integrated into DMNs.

Optimization of Automated Dimethylmethylene Blue Assay for Quantification of Pediatric Urine Samples.

The metachromatic dye dimethylmethylene blue is used to quantify total glycosaminoglycans in urine. Understanding the interaction of dimethylmethylene blue with glycosaminoglycans is pertinent to optimize the assay procedure depending on the type of sample and interpret the findings meaningfully. The present spectrophotometric study determined the optimum sample-to-dye ratio, primary wavelength for measuring absorbance, after studying the interaction of two different chondroitin sulfate species (unfractionated chondroitin sulfate from bovine trachea vs. chondroitin sulfate oligosaccharide with degree of polymerization of 12, from shark cartilage) with dimethylmethylene blue. Respective dye-glycosaminoglycan complexes of the two chondroitin sulfate species showed significantly different absorbance maxima, while that of the chondroitin sulfate oligosaccharide was closer to absorbance maxima of urine glycosaminoglycans. The chondroitin sulfate oligosaccharide showed relatively less stable absorbance readings at higher concentrations in the reaction volume. Furthermore, the chondroitin sulfate reference materials exhibited differences in the linearity of standard curves and hence parallelism. Based on the findings, the method was semiautomated on Beckman Coulter D✕C 700 biochemistry analyzer using the chondroitin sulfate oligosaccharide as the standard. The urine glycosaminoglycan concentration obtained was slightly lower but reasonably close to that obtained through the External Quality Assurance (EQA) scheme administrated by ERNDIM (European Research Network, Inherited Disorders of Metabolism). The findings of the present study can be used to guide the dimethylmethylene blue assay optimization, redevelopment efforts, and harmonization across laboratories. The chondroitin sulfate oligosaccharide is better than the unfractionated chondroitin sulfate from bovine trachea due to its absorbance maxima closer to urine glycosaminoglycans. On the other hand, unfractionated chondroitin sulfate exhibit poor parallelism leading to falsely lower urine glycosaminoglycan levels.

Automated Calibration for Rapid Optical Spectroscopy Sensor Development for Online Monitoring.

An automated platform has been developed to assist researchers in the rapid development of optical spectroscopy sensors to quantify species from spectral data. This platform performs calibration and validation measurements simultaneously. Real-time, in situ monitoring of complex systems through optical spectroscopy has been shown to be a useful tool; however, building calibration models requires development time, which can be a limiting factor in the case of radiological or otherwise hazardous systems. While calibration time can be reduced through optimized design of experiments, this study approached the challenge differently through automation. The ATLAS (Automated Transient Learning for Applied Sensors) platform used pneumatic control of stock solutions to cycle flow profiles through desired calibration concentrations for multivariate model construction. Additionally, the transients between desired concentrations based on flow calculations were used as validation measurements to understand model predictive capabilities. This automated approach yielded an incredible 76% reduction in model development time and a 60% reduction in sample volume versus estimated manual sample preparation and static measurements. The ATLAS system was demonstrated on two systems: a three-lanthanide system with Pr/Nd/Ho representing a use case with significant overlap or interference between analyte signatures and an alternate system containing Pr/Nd/Ni to demonstrate a use case in which broad-band corrosion species signatures interfered with more distinct lanthanide absorbance profiles. Both systems resulted in strong model prediction performance (RMSEP < 9%). Lastly, ATLAS was demonstrated as a tool to simulate process monitoring scenarios (e.g., column separation) in which models can be further optimized to account for day-to-day changes as necessary (e.g., baseline correction). Ultimately, ATLAS offers a vital tool to rapidly screen monitoring methods, investigate sensor fusion, and explore more complex systems (i.e., larger numbers of species).

Utilizing erythrosine B absorption spectrum shifts for quantitative determination of octreotide and bromocriptine in their pure forms and pharmaceutical formulations. Evaluation of the method greenness.

In the present study, two drugs for treating acromegaly are investigated which are either synthetic growth hormone inhibiting hormone (GHIH) or dopamine receptor agonist. Octreotide (OCT) and bromocriptine (BCT) were quantified with a quick, simple, and sensitive spectrophotometric approach in their authentic forms and commercial dosage forms. This approach was based upon formation of ion pair complex between erythrosine B and investigated drugs in an aqueous buffered solution. For OCT determination the higher sensitivity was obtained using ΔA at 525 nm. On the other hand, BCT was estimated using the absorbance at 556 nm. Under optimal conditions for the reaction, construction of calibration curves were performed within concentration range of 0.4-4 µg ml-1 for OCT and 1-9 µg ml-1 for BCT with with low detection limits (0.094 and 0.235 µg ml-1) and low quantitation limits (0.29 and 0.71 µg ml-1) for OCT and BCT respectively. The developed approach was assessed for linearity, accuracy, precision, limits of detection, and limits of quantitation in compliance with ICH validation recommendations. The suggested approaches demonstrated good linearity, as evidenced by determination coefficients (r2) of 0.999 with a slope 0.14 for OCT and 0.9989 with a slope 0.06 for BCT. Additionally, the environmental friendliness of the investigated method was assessed with some of the recent green analytical chemistry metrics.

Rapid ion pair-based surfactant-assisted liquid-liquid microextraction with solidification of floating organic drop for simultaneous spectrophotometric determination of selected anionic dyes in food samples.

Ion pair-based surfactant-assisted liquid-liquid microextraction with solidification of floating organic drops has been developed to extract Allura red (AR), tartrazine (TAR), and fast green (FG) prior to spectrophotometric determination. Cetyltrimethylammonium bromide (CTAB) was employed as ion-pairing agent to enhance the hydrophobic behavior of anionic dyes. 1-undecanol and ethanol were used as the extraction and dispersion solvents, respectively. The dyes were quantitatively extracted in the presence of KCl (0.15 mol L-1) at pH 4.0. The method exhibits wide linearity (15.0-1500.0 µg L-1 for AR, 35.0-2000.0 µg L-1 for TAR, and 3.0-1200.0 µg L-1 for FG) with preconcentration factors of 19.6, 20.1, and 19.9, respectively. The detection limit was 3.7. 9.5, and 0.83 µg L-1 for AR, TAR, and FG, respectively. The relative standard deviation did not exceed 2.1 %. The procedure was applied for the determination of these dyes in food samples.

Ambient light spectrum affects larval Mexican jumping bean moth (Cydia saltitans) behavior despite light obstruction from host seed.

Spectral differences in ambient light can affect animal behavior and convey crucial information about an individual's environment. The ability to perceive and respond to differences in ambient light varies widely by taxa and is shaped by a species' ecology. Mexican jumping bean moths, Cydia saltitans, spend their entire larval period encased in fallen host seeds and contend with potentially lethal environmental temperatures when host seeds are in direct sunlight. We investigate if and how C. saltitans larvae in host seeds respond to lighting conditions associated with these thermal risks. In a temperature-controlled experiment, we identified that larvae demonstrated distinct behavioral ("jumping") responses corresponding to four lighting treatments (white, red, green, and purple), despite extremely minimal light penetration through host seed walls. Red light induced the greatest larval activity (measured by probability of movement and by displacement from origin), suggesting that larvae have mechanisms to perceive low levels of red light and/or to detect subtle increases in heat produced by red/near infrared-biased light spectra, possibly providing them with an early-warning mechanism against thermal stress. Our findings highlight the interplay of environmental lighting, behavior, and potential thermosensory adaptations in a species with a visually constrained environment.

Determination of xanthophylls in egg yolk using a combination of spectrophotometric and diol phase high-performance liquid chromatography methods.

Normal-phase (NP) liquid chromatography is one of the most effective methods for separating isomers with sensitive structural features, including xanthophyll isomers. In this work, reverse-phase (RP) and NP liquid chromatography (LC), with silica gel and diol phase, respectively, were evaluated for the separation of xanthophyll isomers. The results showed that RP LC with monomeric C18 phase not only poorly separate all xanthophyll isomers in egg yolk but also requires additional sample preparation to eliminate triacylglycerols in egg yolk. The diol phase of NP-LC provided the highest efficiency for separating lutein, zeaxanthin, and their cis-isomers with isocratic separation using mobile phases consisting of n-hexane and polar modifiers (such as acetone, methyl tert-butyl ether, or ethyl acetate). To determine the xanthophyll content, peak areas from LC and total absorbance from spectrophotometry measurements were used. The approach was applied to analyze the xanthophylls of nine commercial egg samples. The results revealed that five out of nine analyzed samples contained a high level of canthaxanthin, which contributes to color enhancement but not to prevent age-related macular degeneration. Together, it shows that NP LC with diol phase combined with spectrophotometry is a powerful tool to monitor xanthophylls in eggs.

Assessment of 5-Hydroxymethylfurfural in Food Matrix by an Innovative Spectrophotometric Assay.

Foods contaminants pose a challenge for food producers and consumers. Due to its spontaneous formation during heating and storage, hydroxymethylfurfural (HMF) is a prevalent contaminant in foods rich in carbohydrates and proteins. Colorimetric assays, such as the Seliwanoff test, offer a rapid and cost-effective method for HMF quantification but require careful optimization to ensure accuracy. We addressed potential interference in the Seliwanoff assay by systematically evaluating parameters like incubation time, temperature, and resorcinol or hydrochloric acid concentration, as well as the presence of interfering carbohydrates. Samples were analyzed using a UV-Vis spectrophotometer in scan mode, and data obtained were validated using HPLC, which also enabled quantification of unreacted HMF for assessing the protocol's accuracy. Incubation time and hydrochloric acid percentage positively influenced the colorimetric assay, while the opposite effect was observed with the increase in resorcinol concentration. Interference from carbohydrates was eliminated by reducing the acid content in the working reagent. HPLC analyses corroborated the spectrophotometer data and confirmed the efficacy of the proposed method. The average HMF content in balsamic vinegar samples was 1.97 ± 0.94 mg/mL. Spectrophotometric approaches demonstrated to efficiently determine HMF in complex food matrices. The HMF levels detected in balsamic vinegars significantly exceeded the maximum limits established for honey. This finding underscores the urgent need for regulations that restrict contaminant levels in various food products.

Isolation of Lactic Acid Bacteria Eliminating Trimethylamine (TMA) for Application to Fishery Processing.

Fishy odor of fish flesh (meat) presents a severe problem for marine production. The main cause of fishy odor is trimethylamine (TMA), which increases during storage. It is produced from trimethylamine oxide (TMAO), an osmosis-regulating substance in fish cells that functions by a reduction reaction. Bacterial growth in fish meat increases TMA. Its odor reduces the commercial value of the meat. Technologies for its regulation and elimination are desired. This chapter presents a description of the use of lactic acid to eliminate TMA. The lactic acid is producible safely by bacteria during food processing using picric acid-toluene.A method of eliminating TMA was demonstrated using Lactobacillus plantarum H78. Furthermore, an assay method was explained for reducing TMA in fish meat by fermenting the H78 strain.