Alkylresorcinol detection and identification in archaeological pottery using ultra-high-performance liquid chromatography-quadrupole/Orbitrap mass spectrometry.

RATIONALE: Alkylresorcinols (AR) are cereal-specific biomarkers and have recently been found in archaeological pots. However, their low concentrations and high susceptibility to degradation make them difficult to detect using conventional gas chromatography mass spectrometry (GC/MS). Here we describe the development of a more sensitive liquid chromatography mass spectrometry (LC/MS) method to detect these compounds. METHOD: A method based on the use of ultra-high-performance liquid chromatography (UHPLC) coupled to an Orbitrap mass analyser was established and validated for the detection of low-concentration ARs in pottery. During the preliminary experiments, UHPLC-Q/Orbitrap MS (ultra-high-performance liquid chromatography-quadrupole/Orbitrap mass spectrometry) was demonstrated to be more sensitive, and a wide range of AR homologues in cereal extracts were detected, unlike UHPLC-QTOFMS (ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry) and GC/MS. The developed method was utilised to profile AR homologue distribution in modern cereal samples and reanalyse AR-containing pots from the archaeological site of Must Farm. RESULTS: A highly sensitive LC/MS method with a limit of detection (LOD) of 0.02 µg/g and a limit of quantification (LOQ) of 0.06 µg/g was used to profile ARs in five modern cereal grains. The obtained LOD is 250 times lower than that obtained using the conventional GC/MS approach. AR 21:0 was the most abundant homologue in all four Triticum spp.-einkorn, emmer, Khorasan wheat and common wheat. Meanwhile, AR 25:0 was the predominant homologue in barley, potentially enabling differentiation between wheat and barley. The developed LC/MS-based method was successfully used to analyse ARs extracted from Must Farm potsherds and identified the cereal species most likely processed in the pots-emmer wheat. CONCLUSION: The described method offers an alternative and more sensitive approach for detecting and identifying ARs in ancient pottery. It has been successfully utilised to detect AR homologues in archaeological samples and discriminate which cereal species-wheat and barley-were processed in the pots.

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.

The Effects of a Cultivar and Production System on the Qualitative and Quantitative Composition of Bioactive Compounds in Spring Wheat (Triticum sp.).

Spelt Triticum aestivum L. subsp. spelta (cv. Wirtas), einkorn Triticum monococcum L. (cv. Samopsza) and emmer Triticum dicoccum Schrank (Schuebl) (cv. Plaskurka biala and Plaskurka ciemna) spring wheat cultivars were analyzed and compared to common wheat Triticum aestivum L. subsp. aestivum (cv. Harenda, Kandela, Mandaryna, Serenada, Goplana, Kamelia, Nimfa, Rusalka, Struna, Zadra) cultivated in an organic production system. Moreover, the performance of four common wheat cultivars (cv. Harenda, Kandela, Mandaryna, Serenada) grown in organic, conventional and integrated production systems were compared. The UHPLC-DAD-MS and TLC-DPPH• analyses of specific substances (phenolic acids and alkylresorcinols) were evaluated to ascertain the potential of spring wheat cultivars for promoting human health and suitability for cultivation in an organic production system. The highest yield was observed for the T. aestivum L. subsp. aestivum (modern hull-less) cv. Nimfa (4.45 t/ha), which also demonstrated the lowest resistance to Fusarium spp. infection. Among the contemporary hull-less cultivars, cv. Mandaryna and cv. Harenda exhibited the highest resistance to this pathogen (2.4% and 3.7% of grains infected by Fusarium, respectively), while simultaneously displaying the highest organic phenolic acid content (900.92 and 984.55 µg/g of the grain) and the highest antioxidant potential. It is noteworthy that the cereal hulls of T. monococcum L. (old hulled) (cv. Samopsza) exhibited a markedly elevated content of phenolic acids (approximately 4000 µg/g of the grain). This may have contributed to the reduced incidence of Fusarium infection (9.3% of grains infected) observed in the grains of this cultivar. Furthermore, the hulls proved to be a rich source of phenolics with high antioxidant activity, which is beneficial for human and animal health.

Highly sensitive and rapid detection of resorcinol by forming fluorescent azamonardine with dopamine.

We propose a sensitive, selective, and rapid fluorescent assay for detecting resorcinol (RC) based on its specific chemical reaction with dopamine. Under alkaline condition, RC would react with dopamine to yield fluorescent azamonardine, which emits strong blue fluorescence and has a superior excitation wavelength at 416 nm and an emission wavelength at 461 nm. The azamonardine with a molecular weight of 258.1 confirmed by ICP-MS has a quantum yield of 71.3%. The reaction is completed within 1 min showing great potential for point-of-care testing. This assay showed high sensitivity and had a good relationship between fluorescent intensity at 461 nm and RC concentration (I461 = 106.4 + 93.6*CRC; R2 = 0.9904) over the range of 0-40 µM. More importantly, the assay showed a prominent anti-interference from various substances and even can distinguish RC from its isomers, o-dihydroxybenzene and p-dihydroxybenzene. Finally, our assay successfully quantified RC contents in wheat powder and hair dyes with high accuracy.

Spectrophotometric determination of nitrate in small volume of seawater samples using a simple resorcinol method.

A simple resorcinol method to determine nitrate (NO3-) in seawater using a microplate reader with a 48-well plate was established. The method involved the nitration of resorcinol in sulfuric acid to form a pink product that was detected at 505 nm. Reagent concentrations were optimized, and the effect of salinity on NO3- determination was investigated. The detection limit of this method was 0.8 µM, while the upper limit of the linear range was 100 µM. The recoveries ranged from 91.5 to 109.7% for spiked seawater samples with different salinities. The proposed method was compared with two reference methods, and the results revealed a good correlation. Compared to conventional methods that require the preparation of reactants, the proposed method used aqueous solutions as reagents for the reaction, which was simpler and more convenient. Compared to the methods that used organic solvents for the direct determination of NO3-, the proposed method was suitable for estuarine and coastal water samples with large salinity variations. All results indicated that the proposed method can satisfy the requirements of laboratory analysis and demonstrate high application potential for use in field determination.

An efficient differential sensing strategy for phenolic pollutants based on a nanozyme with polyphenol oxidase activity.

To realize the efficient differential sensing of phenolic pollutants in sewage, a novel sensing strategy was successfully developed based on a nanozyme (GMP-Cu) with polyphenol oxidase activity. Phenolic pollutants can be oxidized using GMP-Cu, and the oxidation products reacts subsequently with 4-aminoantipyrine to produce a quinone-imine compound. The absorption spectra of final quinone-imine products that resulted from different phenolic pollutants showed obvious differences, which were due to the interaction difference between GMP-Cu and phenolic pollutants, as well as the different molecular structures of the quinone-imine products from different phenolic pollutants. Based on the difference in the absorption spectra, a novel differential sensing strategy was developed. A genetic algorithm was used to select the characteristic wavelengths at different enzymatic reaction times. Hierarchical cluster analysis and PLS-DA algorithms were utilized for the discriminant sensing of seven representative phenolic pollutants, including hydroquinone, resorcinol, catechol, resorcinol, phenol, p-chlorophenol, and 2,4-dichlorophenol. A scientific wavelength selection algorithm and a recognition algorithm resulted in the successful identification of phenolic pollutants in sewage with a discriminant accuracy of 100%, and differentiation of the phenolic pollutants regardless of their concentration. These results indicated that a sensing strategy can be used as an effective tool for the efficient identification and differentiation of phenolic pollutants in sewage.

Synthesis of morphology-controlled N-doped porous carbon for simultaneous electrochemical sensing of dihydroxybenzene isomers.

Different morphology of N-doped carbon materials, including three-dimensional interconnected N-doped hierarchically porous carbon networks (3D-NC), two-dimensional ultrathin porous carbon nanosheets (2D-NC), and bulk N-doped carbon with micron size (bulk-NC), was easily prepared by using NaCl crystal templates-assisted strategy. Compared with bare glassy carbon, bulk-NC, and 2D-NC, the as-synthesized 3D-NC exhibits excellent electrochemical activity toward the oxidation and sensing of three kinds of common environmental pollutants dihydroxybenzene isomers (hydroquinone (HQ), catechol (CC), and resorcinol (RS)). The impressive electrochemical activity of 3D-NC can be interpreted by its large specific surface area, continuous network-like morphology, superior electro-catalytic ability, and strong accumulation efficiency. Differential pulse voltammetry (DPV) test showed the 3D-NC-modified electrode exhibited three well-separated oxidation peaks at 0.05 V, 0.14 V, and 0.45 V vs. saturated calomel electrode (SCE) for HQ, CC, and RS, and their detection limits were evaluated to be as low as 0.0044, 0.012, and 0.016 mg L-1, respectively. Finally, a novel electrochemical analytical platform is successfully fabricated for the simultaneous monitoring of hydroquinone, catechol, and resorcinol with high sensitivity. When used for real wastewater samples analysis, recovery ratio ranging from 94 to 108% with lower than 5% of relative standard deviation (RSD) values was achieved. This work proves a facile strategy to prepare morphology-controlled N-doped carbon-based material and demonstrates its high application potential for environmental monitoring and electrochemical analysis.

Fluorescence "turn-on" of silicon-containing nanoparticles for the determination of resorcinol.

A fluorescent nanosensor based on silicon-containing nanoparticles (Si CNPs) with green fluorescence (FL) was prepared by one-step method. The prepared Si CNPs emitted green FL at 470 nm under the excitation at 350 nm. The FL signal of Si CNPs reveals an obvious enhancement in the presence of resorcinol (RC), due to the passivation of surface trap states of Si CNPs via the binding of OH group of RC with the NH group of Si CNPs, which allowed the formation of new radiative electron-hole recombination centers. This was confirmed by some analytical experiments performed on zeta potential, FL lifetime steady state, and the FTIR spectra. Most importantly, this nanosensor could selectively determine RC with high sensitivity and without interference from hydroquinone (HQ) and catechol (CT) as RC isomers. RC was detected in the linear range 0.05-40 µM, with a detection limit of 0.012 µM. The synthesized nanosensor was applied to the determination of RC in fresh fruit juice and water samples. The collected results confirmed the feasibility of our approach with high accuracy.

Simplified method for the measurement of plasma alkylresorcinols: Biomarkers of whole-grain intake.

RATIONALE: Consumption of whole grains is negatively associated with cardiovascular disease (CVD) risk but quantification of whole-grain intake is challenging. Alkylresorcinols (ARs) are biomarkers of whole-grain intake. Current methods for AR quantification involve a time-consuming multi-step separation process that hampers applicability in large-scale studies. METHODS: We developed a streamlined method to quantify ARs in human plasma based on protein precipitation and direct injection into an ultra-high-performance liquid chromatograph coupled to a quadrupole time-of-flight mass spectrometer operating in atmospheric pressure chemical ionization negative ion mode. RESULTS: Separation of five major ARs was achieved, with linearity in the 5 to 550 nmol/L range and a lower limit of detection (LOD) of 0.5 nmol/L and quantification (LOQ) of 5 nmol/L. The within-run and between-run precision and accuracy were below 15%, and recoveries above 90%. Once validated, the method was applied to measure concentrations of plasma ARs in subjects who participated in a randomized, crossover trial evaluating the effect of carbohydrate type on CVD risk factors. The unrefined carbohydrate diet with the highest fiber content resulted in the highest plasma AR concentration (93 ± 78 nmol/L), and was significantly different (p <0.01) from lower fiber diets (18 ± 26 nmol/L and 19 ± 26 nmol/L, simple and unrefined carbohydrate, respectively). CONCLUSIONS: This method offers a simplified approach to measure concentrations of plasma ARs as an objective biomarker of whole-grain intake that can be applied to large-scale cohort studies.

Colorimetric paper bioassay by horseradish peroxidase for the detection of catechol and resorcinol in aqueous samples.

Phenolic compounds such as catechol and resorcinol are toxic and persistent pollutants in the aqueous environment. Detection procedures such as chromatographic and spectrophotometric methods are time-consuming and require sophisticated instruments with skilled manpower. Development of a simple, cost effective, portable and disposable paper based biosensor could be a better alternative to the conventional methods. The present study attempted to develop a paper based biosensor by immobilizing horseradish peroxidase enzyme to detect catechol and resorcinol in aqueous samples. Horseradish peroxidase catalyzes the oxidation of phenolic compounds to semiquinones, which on reaction with a chromogen, 3-methyl 2-benzothiazolinone hydrazine (MBTH) gives faint pink to red color depending on the compound and its concentration in the sample is the basis for biosensing application. Different methods of enzyme immobilization on filter paper like physical adsorption, covalent coupling, and polysaccharide entrapment were executed. The performance of the various enzyme immobilization methods was evaluated by analyzing the developed color intensity using ImageJ software. Entrapment technique is the most effective method of immobilizing enzyme on the filter paper that produces the highest color intensity with better stability. The visible limit of detection (LoD) was observed as 0.45 mM (50 mg/L) for catechol and 0.09 mM (10 mg/L) for resorcinol in aqueous samples.

Dietary options and behavior suggested by plant biomarker evidence in an early human habitat.

The availability of plants and freshwater shapes the diets and social behavior of chimpanzees, our closest living relative. However, limited evidence about the spatial relationships shared between ancestral human (hominin) remains, edible resources, refuge, and freshwater leaves the influence of local resources on our species' evolution open to debate. Exceptionally well-preserved organic geochemical fossils--biomarkers--preserved in a soil horizon resolve different plant communities at meter scales across a contiguous 25,000 m(2) archaeological land surface at Olduvai Gorge from about 2 Ma. Biomarkers reveal hominins had access to aquatic plants and protective woods in a patchwork landscape, which included a spring-fed wetland near a woodland that both were surrounded by open grassland. Numerous cut-marked animal bones are located within the wooded area, and within meters of wetland vegetation delineated by biomarkers for ferns and sedges. Taken together, plant biomarkers, clustered bone debris, and hominin remains define a clear spatial pattern that places animal butchery amid the refuge of an isolated forest patch and near freshwater with diverse edible resources.

Characterization and topical delivery of phenylethyl resorcinol.

OBJECTIVE: Phenylethyl resorcinol (PR) has been used widely in the personal care industry as a novel skin lightening ingredient. Surprisingly, there is only limited information describing the physicochemical properties of this active. Therefore, the primary objective of this study was to perform a comprehensive characterization of PR. A secondary objective was to investigate the delivery of this molecule to mammalian skin. METHODS: Phenylethyl resorcinol was characterized using differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and nuclear magnetic resonance (NMR). A new high-performance liquid chromatographic (HPLC) method for analysis of PR was developed and validated. The log P (octanol water partition coefficient), value, solubility and short-term stability of PR in a series of vehicles were also determined using HPLC. The evaporation of the selected vehicles was examined using dynamic vapour sorption (DVS). The permeation profiles of PR were investigated under finite dose conditions in porcine and human skin. RESULTS: The melting point of PR was determined to be 79.13 °C and the measured log P (octanol water partition coefficient) at 21 °C was 3.35 ± 0.03. The linearity of the HPLC analytical method was confirmed with an r2 value of 0.99. Accuracy of the method was evaluated by average recovery rates at three tested concentrations, and the values ranged from 99 to 106%. The limit of detection (LOD) and limit of quantification (LOQ) were 0.19 and 0.57 µg mL-1 , respectively. The solubility of PR in PG, DMI, glycerol was within the range of 367 to 877 mg mL-1 . The stability of PR in tested solvents was also confirmed by the 72 h stability studies. From the DVS studies, 70-125% of applied formulations were recovered at 24 h. The permeation through porcine skin at 24 h ranged from 4 to 13 µg cm-2 , while the corresponding amounts of PR delivered through human skin were 2 to 10 µg cm-2 . CONCLUSION: The physicochemical properties of PR confirm it is suitable for dermal delivery. In this study, propylene glycol was the most promising vehicle for PR delivery to human skin. Future work will expand the range of vehicles studied and explore the percutaneous absorption from more complex formulations.

OBJECTIF: Le phényléthyl résorcinol (PR) est largement utilisé dans le secteur des soins personnels comme ingrédient éclaircissant pour la peau. Pour autant, on ne dispose que d'informations limitées concernant les propriétés physicochimiques de ce principe actif. C'est pourquoi cette étude avait pour objectif principal de réaliser une caractérisation exhaustive du PR. Son objectif secondaire était d'étudier l'administration de cette molécule à la peau de mammifères. MÉTHODES: Le phényléthyl résorcinol a été caractérisé par calorimétrie différentielle à balayage (CDB), analyse thermogravimétrique (ATG) et par résonance magnétique nucléaire (RMN). Pour analyser le PR, une nouvelle méthode de chromatographie liquide à haute performance (CLHP) a été développée et validée. On s'est servi de la CLHP pour déterminer les propriétés suivantes du PR : log P (coefficient de partage octanol/eau), valeur, solubilité et stabilité à court terme du PR dans plusieurs véhicules. L'évaporation des véhicules sélectionnés a été examinée par sorption de vapeur dynamique (DVS). Les profils de perméabilité du PR ont été étudiés dans des conditions de dose finie dans des peaux porcine et humaine. RÉSULTATS: On a pu déterminer que le point de fusion du PR était de 79,13 °C et le log P (coefficient de partage octanol/eau) à 21 °C était de 3,35 ± 0.03. La linéarité de la méthode analytique de la CLHP a été confirmée avec une valeur r2 de 0,99. L'exactitude de la méthode a été évaluée par les taux moyens de récupération à trois concentrations testées, avec des valeurs résultantes comprises entre 99 et 106 %. La limite de détection (LD) et la limite de quantification (LQ) ont été déterminées à 0,19 et 0,57 µg/ml_ 1, respectivement. La solubilité du PR dans le PG, le DMI et le glycérol reste dans une plage comprise entre 367 et 877 mg/ml _ 1. La stabilité du PR dans les solvants testés a également pu être confirmée par les études de stabilité à 72 h. Parmi les formulations appliquées lors des études de DVS, 70 à 125 % de celles-ci ont été récupérées à 24 h. La pénétration par la peau porcine à 24 h était comprise entre 4 et 13 µg/cm_ 2, tandis que les quantités de PR correspondantes délivrées à travers la peau humaine étaient de 2 à 10 µg/cm_ 2. CONCLUSION: Les propriétés physicochimiques du PR confirment qu'il est adapté à l'administration dermique. Dans le cadre de cette étude, le propylène glycol est apparu comme le véhicule le plus prometteur pour l'administration de PR dans la peau humaine. De futurs travaux étudieront davantage de véhicules et examineront l'absorption percutanée lors de l'emploi de formulations plus complexes.

Evaluation of alkylresorcinols in adipose tissue biopsies as a long-term biomarker of whole-grain wheat and rye intake in free-living Swedish men and women.

OBJECTIVE: Wheat and rye, the most consumed whole grains (WG) in the Nordic countries, contain alkylresorcinols (AR) in their bran. AR concentrations in human adipose tissue might reflect long-term WG rye and wheat intake. We aimed to evaluate AR concentrations in adipose tissue biopsies as a long-term biomarker of WG wheat and rye intake in free-living Swedish men and women. DESIGN: Cross-sectional study. AR concentrations in adipose tissue biopsies were analysed and compared with long-term WG intake assessed by three FFQ (repeated over a period of 14 years in men, 17 years in women) and with plasma AR concentrations. SETTING: The Cohort of Swedish Men between 1997 and 2010 and the Swedish Mammography Cohort between 1987 and 2003, Sweden. SUBJECTS: Men (n 149) and women (n 109). RESULTS: Long-term WG rye intake estimated with repeated FFQ correlated (r=0·31-0·41, P<0·01) with adipose-tissue AR concentrations, while WG wheat intake correlated only weakly (r=0·17-0·33, P<0·05). Total AR concentration in adipose tissue was 61 % lower in women than in men at similar energy-adjusted WG wheat and rye intakes, but plasma concentrations were similar. AR concentrations in adipose tissue correlated well with plasma concentrations (r=0·49-0·81, P<0·001). CONCLUSIONS: AR in adipose tissue reflected long-term WG rye but not WG wheat intake, probably due to poor precision in estimating WG wheat intake by FFQ. AR in adipose tissue appears promising as a biomarker of long-term WG rye intake but should be adjusted for sex.

Simultaneous removal of dihydroxybenzenes and toxicity reduction by Penicillium chrysogenum var. halophenolicum under saline conditions.

The dihydroxybenzenes are widely found in wastewater and usually more than one of these aromatic compounds co-exist as pollutants of water resources. The current study investigated and compared the removal efficiency of hydroquinone, catechol and resorcinol in binary substrate systems under saline conditions by Penicillium chrysogenum var. halophenolicum, to clarify the potential of this fungal strain to degrade these aromatic compounds. Since P. chrysogenum is a known penicillin producer, biosynthetic penicillin genes were examined and antibiotic was quantified in mono and binary dihydroxybenzene systems to elucidate the carbon flux of dihydroxybenzenes metabolism in the P. chrysogenum var. halophenolicum to the secondary metabolism. In binary substrate systems, the three assayed dihydroxybenzene compounds were found to be co-metabolized by fungal strain. The fungal strain preferentially degraded hydroquinone and catechol. Resorcinol was degraded slower and supports higher antibiotic titers than either catechol or hydroquinone. Dihydroxybenzenes were faster removed in mixtures compared to mono substrate systems, except for the case of hydroquinone. In this context, the expression of penicillin biosynthetic gene cluster was not related to the removal of dihydroxybenzenes. Penicillin production was triggered simultaneously or after dihydroxybenzene degradation, but penicillin yields, under these conditions, did not compromise dihydroxybenzene biological treatment. To investigate the decrease in dihydroxybenzenes toxicity due to the fungal activity, viability tests with human colon cancer cells (HCT116) and DNA damage by alkaline comet assays were performed. For all the conditions assays, a decrease in saline medium toxicity was observed, indicating its potential as detoxification agent.

Application of hydrolases and probiotic Pediococcus acidilactici BaltBio01 strain for cereal by-products conversion to bioproduct for food/feed.

The aim of this study was to apply the enzymatic treatment and fermentation by Pediococcus acidilactici BaltBio01 strain for industrial cereal by-products conversion to food/feed bioproducts with high amount of probiotic lactic acid bacteria (LAB). LAB propagated in potato media and spray-dried remained viable during 12 months (7.0 log10 cfu/g) of storage and was used as a starter for cereal by-products fermentation. The changes of microbial profile, biogenic amines (BAs), mycotoxins, lactic acid (L+/D-), lignans and alkylresorcinols (ARs) contents in fermented cereal by-product were analysed. Cereal by-products enzymatic hydrolysis before fermentation allows to obtain a higher count of LAB during fermentation. Fermentation with P. acidilactici reduce mycotoxins content in fermented cereal by-products. According to our results, P. acidilactici multiplied in potato juice could be used for cereal by-products fermentation, as a potential source to produce safer food/feed bioproduct with high amount of probiotic LAB for industrial production.

Comprehensive Characterisation of n-Alkylresorcinols and Other Lipid Constituents of Mercurialis tomentosa L. from Alicante, Spain.

Mercurialis tomentosa L. has been used in Spanish ethnomedicine. In the present study the first phytochemical characterisation of a lipid fraction from M. tomentosa was performed. The CHCl3 extraction of aerial parts from M. tomentosa and GC/MS investigations revealed the occurrence of cuticular lipid and wax constituents, like long chain n-alcohols and n-aldehydes (C22  - C30 ), besides several aromatic constituents, i.e., phenylpropanoids and n-alkylresorcinols. The latter were further purified by CC and analysed by LC/MSn . In contrast to other Mercurialis species, i.e., M. annua, M. perennis, which exclusively contain 5-n-alkylresorcinols (1a - j, Cn ), mainly 5-n-alkyl-2-methylresorcinols (2a - j, Cn *) with side chain lengths of C15  - C25 were found in M. tomentosa, in addition to 1a - j. Thus, the latter compounds may be utilised for analytical characterisation and authentication of M. tomentosa based on fingerprinting methods. For structure elucidation a novel facile total synthesis of one representative 5-n-alkyl-2-methylresorcinol homologue (2d, C19 *) was developed, starting with a Grignard reaction from a substituted benzoic acid chloride (19). The compound obtained by synthesis was identical to the natural product 2d in terms of its chromatographic and spectroscopic features. Futhermore, 2d exhibited satisfactory DPPH free radical scavenging activity (IC50  = 37.8 µm) when compared to trolox (IC50  = 21.0 µm), corroborating the antioxidant features of these amphipathic molecules.

Development and validation of an ultraviolet-visible spectrophotometric method for determination of phenylethyl resorcinol in new topical nanoemulsions.

OBJECTIVE: To develop a rapid, simple and efficient ultraviolet-visible (UV-vis) spectrophotometric method for the quantification of phenylethyl resorcinol (PR), a potent skin-lightening agent, incorporated into new topical nanoemulsions, and to validate this method according to International Commission for Harmonization (ICH) guidelines. METHODS: UV-vis spectrophotometric method for quantification of PR in new topical nanoemulsions was developed and validated in terms of specificity, linearity, sensitivity, precision and accuracy. The method was applied to determine PR content (extraction recoveries) in the nanoemulsions and entrapment efficiencies. RESULTS: The calibration curve for PR was linear in the range of 10-60 µg mL-1 , with a determination coefficient (r2 ) which is higher than 0.999. The limit of detection (LOD) and limit of quantitation (LOQ) were 1.55 and 4.69 µg mL-1 , respectively. The method was shown to be specific, precise [intraday, interday and reproducibility levels relative standard deviation < 3%] and accurate (between 91.90 ± 1.28% and 104.73 ± 0.60%). The extraction recoveries of PR in nanoemulsions were 97.07%, 96.64% and 103.54% at concentrations of 3, 6 and 9 mg mL-1 , respectively. The entrapment efficiencies were nearly 100%, which agrees with the oil core location of PR in nanoemulsions. CONCLUSION: This developed method was confirmed to be rapid, simple, cost-effective, specific, precise, accurate and suitable for determination of PR content in nanoemulsions and entrapment efficiencies.

Evaluation of the potential of alkylresorcinols as superoxide anion scavengers and sox-regulon modulators using nitroblue tetrazolium and bioluminescent cell-based assays.

The antioxidant activities of five alkylresorcinol (AR) homologs with alkyl chains of 1, 3, 5 6 and 12 carbon atoms were studied using molecular and cellular assays for superoxide anions (O2.-). The effect of ARs as superoxide anion scavengers was assessed using the photochemical reaction of spontaneous photo-reduced flavin re-oxidation. In this system, ARs reaction with O2.- produced dye derivatives, as C6- and C12-AR prevented the O2.--induced conversion of nitroblue tetrazolium into formazan in AR-containing mixtures. The influence of ARs on soxS gene expression and bacterial cell viability was studied with the luminescent Escherichia coli K12 MG1655 psoxS'::luxCDABE-AmpR strain, showing low basal light emission. This increased significantly during paraquatinduced oxidative stress as a consequence of the simultaneous transcription of soxS-gene and lux-gene fusion. ARs with alkyl chains containing 5-12 carbon atoms at concentrations of 0.1-1.0 µM weakly induced soxS-gene expression, whereas 1-10 mM repressed it. This respectively increased or decreased the bacterial cell resistance to O2.- -related oxidative stress. AR derivatives lost their protective activity from reactions with superoxide anions, which required increased soxS gene expression for cell viability. These results show the dual nature of ARs, which possess direct antioxidant properties and the ability to indirectly regulate the activity of cellular antioxidative defense mechanisms.

Indicator displacement-based colorimetric assay for dibutyl phthalate in pharmaceutical products with titanium(IV)-pyridylazo resorcinol (PAR).

Taking advantage of the competitive binding affinity towards Ti(IV) between 4-(2-pyridylazo) resorcinol (PAR) and phthalate, a simple indicator displacement (ID)-based colorimetric assay was designed for indirect determination of a well-known phthalic acid ester, dibutyl phthalate (DBP). The indicator PAR and Ti(IV) formed a purplish-red-colored Ti(IV)-PAR complex (λmax = 540 nm) at a 1:1 ratio. In the presence of pre-hydrolyzed DBP, colorless complex formation of phthalate ion (emerging from alkaline hydrolysis of DBP) with Ti(IV) resulted in a hypsochromic shift in absorbance maximum, accompanying a color change from purplish-red to yellowish-orange (λmax = 390 nm) by the release of PAR from Ti(IV)-PAR system. Based on this mechanism, the linear response range of the system for DBP was found to lie between 0.16 and 0.37 mmol L-1 with an experimental detection limit of 11.6 µmol L-1. The recommended Ti(IV)-PAR system was successfully applied to DBP-containing pharmaceutical products (as real sample) after a simple clean-up process for removing possible water-soluble interferents. The analytical results obtained from the recommended method (by applying the standard addition approach) and the reference liquid chromatography-tandem mass spectrometric (LC-MS/MS) method were statistically compared using DBP-extract of the drug samples. Consequently, a simple and selective colorimetric ID strategy was proposed for the analysis of DBP in pharmaceuticals for the first time.

Molecularly Imprinted Electrochemical Sensor Based on α-Cyclodextrin Inclusion Complex and MXene Modification for Highly Sensitive and Selective Detection of Alkylresorcinols in Whole Wheat Foods.

Authenticating whole wheat foods poses a significant challenge for both the grain industry and consumers. Alkylresorcinols (ARs), serving as biomarkers of whole wheat, play a crucial role in assessing the authenticity of whole wheat foods. Herein, we introduce a novel molecularly imprinted electrochemical sensor with modifications involving a molecularly imprinted polymer (MIP) and MXene nanosheets, enabling highly sensitive and selective detection of ARs. Notably, we specifically chose 5-heneicosylresorcinol (AR21), the predominant homologue in whole wheat, as the template molecule. α-Cyclodextrin and acrylamide served as dual functional monomers, establishing a robust multiple interaction between the MIP and AR21. As a result, the sensor exhibited a wide linear range of 0.005 to 100 µg·mL-1 and a low detection limit of 2.52 ng·mL-1, demonstrating exceptional selectivity and stability. When applied to commercial whole wheat foods, the assay achieved satisfactory recoveries and accuracy, strongly validating the practicality and effectiveness of this analytical technique.