Bisphenol A and its analogues: from their occurrence in foodstuffs marketed in Europe to improved monitoring strategies-a review of published literature from 2018 to 2023.

In this review article, the research works covering the analytical determination of bisphenol A (BPA) and its structural analogues published from 2018 to present (February 2024) were examined. The review offers an overview of the concentration levels of these xenoestrogens in food and beverages, and discusses concerns that these may possibly pose to the human health and scrutinises, from an analytical perspective, the main biomonitoring approaches that are applied. This comes as a natural evolution of a previous review that covered the same topic but in earlier years (up to 2017). As compared to the past, while the volume of published literature on this topic has not necessarily decreased, the research studies are now much more homogeneous in terms of their geographical origin, i.e., Southern Europe (mainly Italy and Spain). For this reason, an estimated daily intake of the European population could not be calculated at this time. In terms of the analytical approaches that were applied, 67% of the research groups exploited liquid chromatography (LC), with a detection that was prevalently (71%) afforded by mass spectrometry, with over one-fourth of the research teams using fluorescence (26%) and a minority (3%) detecting the analytes with diode array detection. One-third of the groups used gas chromatography (GC)-mass spectrometry achieving comparatively superior efficiency as compared to LC. Derivatisation was performed in 59% of the GC studies to afford more symmetrical signals and enhanced sensitivity. Although the contamination levels are well below the threshold set by governments, routinely biomonitoring is encouraged because of the possible accumulation of these contaminants in the human body and of their interplay with other xenoestrogens.

Dermocosmetic evaluation of a nutricosmetic formulation based on Curcuma.

Endogenous and exogenous factors can alter the skin layer and appearance, determining skin aging. The extracts and isolated molecules from food matrixes can be used to formulate "healthy" antiaging cosmetics. Two different cosmetic approaches can be used to achieve the antiaging effect. It is possible to use topical products based on food extract (cosmeceutical approach) or take a food supplement and apply a topical cosmetic product based on food extract on the surface to be treated (nutricosmetic approach). This work evaluated in vivo the antiaging potential of a nutricosmetic formulation (cream + food supplement) and a cosmeceutical cream based on Curcuma. The choice of the commercial Curcuma extract to be used for experimental purposes was based on the curcuminoid content determined by an HPLC test. Curcuminoids are the bioactive compounds responsible for Curcuma's antioxidant and antiinflammatory properties. Their levels in Curcuma extracts vary according to the storage condition, variety, and pedoclimatic cultivation conditions. The Tewameter® TM300 was used to evaluate the Trans Epidermal Water Loss (TEWL), the Corneometer® CM 825 to determine the moisturizing effect, the Cutometer® to estimate the skin firmness and elasticity, the Dermascan to assess the collagen index, and the Visioface® 1000D to evaluate the wrinkles. The nutricosmetic product showed potential as moisturizing, anti-age, and anti-wrinkle action better than the cosmeceutical product alone.

Investigating the Effect of Surface Hydrophilicity on the Destiny of PLGA-Poloxamer Nanoparticles in an In Vivo Animal Model.

This study aimed to examine the impact of different surface properties of poly(lactic-co-glycolic) acid (PLGA) nanoparticles (P NPs) and PLGA-Poloxamer nanoparticles (PP NPs) on their in vivo biodistribution. For this purpose, NPs were formulated via nanoprecipitation and loaded with diphenylhexatriene (DPH), a fluorescent dye. The obtained NPs underwent comprehensive characterization, encompassing their morphology, technological attributes, DPH release rate, and thermodynamic properties. The produced NPs were then administered to wild-type mice via intraperitoneal injection, and, at scheduled time intervals, the animals were euthanized. Blood samples, as well as the liver, lungs, and kidneys, were extracted for histological examination and biodistribution analysis. The findings of this investigation revealed that the presence of poloxamers led to smaller NP sizes and induced partial crystallinity in the NPs. The biodistribution and histological results from in vivo experiments evidenced that both, P and PP NPs, exhibited comparable concentrations in the bloodstream, while P NPs could not be detected in the other organs examined. Conversely, PP NPs were primarily sequestered by the lungs and, to a lesser extent, by the kidneys. Future research endeavors will focus on investigating the behavior of drug-loaded NPs in pathological animal models.

Recent Advances in Natural Polyphenol Research.

Polyphenols are secondary metabolites produced by plants, which contribute to the plant's defense against abiotic stress conditions (e.g., UV radiation and precipitation), the aggression of herbivores, and plant pathogens. Epidemiological studies suggest that long-term consumption of plant polyphenols protects against cardiovascular disease, cancer, osteoporosis, diabetes, and neurodegenerative diseases. Their structural diversity has fascinated and confronted analytical chemists on how to carry out unambiguous identification, exhaustive recovery from plants and organic waste, and define their nutritional and biological potential. The food, cosmetic, and pharmaceutical industries employ polyphenols from fruits and vegetables to produce additives, additional foods, and supplements. In some cases, nanocarriers have been used to protect polyphenols during food processing, to solve the issues related to low water solubility, to transport them to the site of action, and improve their bioavailability. This review summarizes the structure-bioactivity relationships, processing parameters that impact polyphenol stability and bioavailability, the research progress in nanocarrier delivery, and the most innovative methodologies for the exhaustive recovery of polyphenols from plant and agri-waste materials.

Parabens Permeation through Biological Membranes: A Comparative Study Using Franz Cell Diffusion System and Biomimetic Liquid Chromatography.

Parabens (PBs) are used as preservatives to extend the shelf life of various foodstuffs, and pharmaceutical and cosmetic preparations. In this work, the membrane barrier passage potential of a subset of seven parabens, i.e., methyl-, ethyl-, propyl- isopropyl, butyl, isobutyl, and benzyl paraben, along with their parent compound, p-hydroxy benzoic acid, were studied. Thus, the Franz cell diffusion (FDC) method, biomimetic liquid chromatography (BLC), and in silico prediction were performed to evaluate the soundness of both describing their permeation through the skin. While BLC allowed the achievement of a full scale of affinity for membrane phospholipids of the PBs under research, the permeation of parabens through Franz diffusion cells having a carbon chain > ethyl could not be measured in a fully aqueous medium, i.e., permeation enhancer-free conditions. Our results support that BLC and in silico prediction alone can occasionally be misleading in the permeability potential assessment of these preservatives, emphasizing the need for a multi-technique and integrated experimental approach.

Monitoring of Pollutants Content in Bottled and Tap Drinking Water in Italy.

The concentration levels of thirteen organic pollutants and selected heavy metals were investigated in 40 plastics bottled and tap water samples. Some of the selected contaminants have an ascertained or suspected endocrine disrupting activity, such as Bisphenol A (BPA) and its analogs, and Bis 2-ethylhexyl phthalate (DEHP), which are used by industries as plasticizers. The most frequently detected pollutants were Bisphenol AF (BPAF) (detection frequency (DF) = 67.5%, mean 387.21 ng L-1), DEHP (DF = 62.5%, mean 46.19 µg L-1) and BPA (DF = 60.0%, mean 458.57 ng L-1), with higher concentration levels found in tap waters. Furthermore, a possible level of exposure to thirteen pollutants via drinking water intake was calculated. Our findings show that, even though the occurrence of contaminants and heavy metals in drinking waters does not pose an immediate, acute health risk for the population, their levels should be constantly monitored and "hard-wired" into everyday practice. Indeed, the health impact to the continuous and simultaneous intake of a huge variety of xenobiotics from various sources by humans is complex and still not fully understood.

Phenylalanine Butyramide Is a New Cosmetic Ingredient with Soothing and Anti-Reddening Potential.

Human skin is colonized by diverse commensal microbes, making up the skin microbiota (SM), contributing to skin integrity and homeostasis. Many of the beneficial effects aroused by the SM are exerted by microbial metabolites such as short-chain fatty acids (SCFAs), including butyric acid. The SCFAs can be used in cosmetic formulations against skin diseases to protect SM by preserving and/or restoring their natural balance. Unpleasant sensorial properties and unfavorable physico-chemical properties of butyrate strongly limit its cosmetic use. In contrast, some butyrate derivatives, including phenylalanine butyramide (C13H18N2O2, FBA), a solid form of butyric acid, are odorless while retaining the pharmacokinetic properties and safety profile of butyric acid. This study assessed the FBA's permeation across the skin and its soothing and anti-reddening potential to estimate its cosmetic application. The dosage method used to estimate FBA's levels was validated to be sure of analytical results. The FBA diffusion tests were estimated in vitro using a Franz-type vertical diffusion cell. The soothing action was evaluated in vivo by Colorimeter CL400, measuring the erythema index. The results suggest that the FBA represents an innovative way to exploit the benefits of butyric acid in the cosmetic fields since it cannot reach the bloodstream, is odorless, and has a significative soothing action (decrease the erythema index -15.7% after 30', and -17.8% after 60').

Simultaneous determination of fifteen multiclass organic pollutants in human saliva and serum by liquid chromatography-tandem ultraviolet/fluorescence detection: A validated method.

A quick and inexpensive validated method, based on sample treatment by liquid-liquid microextraction followed by liquid chromatography (LC) coupled with ultraviolet tandem fluorescence detection is proposed for the determination of 15 multiclass pollutants both in serum and in saliva, as a simple and easy to draw matrix. The method was set up and validated according to European guidelines. The compounds of interest include some endocrine-disrupting chemicals (i.e. bisphenol A, bisphenol B, bisphenol E, bisphenol F, bisphenol AF, bisphenol A diglycidyl ether, bisphenol M, diethylhexyl phthalate, monoethylhexyl phthalate, triclosan and 4-nonylphenol), as well as other pollutants belonging to the class of volatile organic compounds (2-chlorophenol, 1,2 dichlorobenzene, 1,2,4,5-tetrachlorobenzene). The limits of quantifications ranged from 2.28 × 10-3 µg mL-1 (bisphenol A diglycidyl ether) to 6.29 µg mL-1 (diethylhexyl phthalate), while those of detection ranged from 0.068 × 10-3 µg mL-1 (bisphenol A diglycidyl ether) to 1.031 µg mL-1 (diethylhexyl phthalate). To test method suitability, it was applied to real saliva and serum samples of healthy human volunteers and was found to meet the demands of the laboratories handling simple and relatively inexpensive equipment for screening oriented at rapid and reliable contamination assessment of a population.

Plasma p-cresol lowering effect of sevelamer in non-dialysis CKD patients: evidence from a randomized controlled trial.

BACKGROUND: The accumulation of p-cresol, a metabolic product of aromatic amino acids generated by intestinal microbiome, increases the cardiovascular risk in chronic kidney disease (CKD) patients. Therefore, therapeutic strategies to reduce plasma p-cresol levels are highly demanded. It has been reported that the phosphate binder sevelamer (SEV) sequesters p-cresol in vitro, while in vivo studies on dialysis patients showed controversial results. Aim of our study was to evaluate the effect of SEV on p-cresol levels in non-dialysis CKD patients. METHODS: This was a single-blind, randomized placebo-controlled trial (Registration number NCT02199444) carried on 69 CKD patients (stage 3-5, not on dialysis), randomly assigned (1:1) to receive either SEV or placebo for 3 months. Total p-cresol serum levels were evaluated at baseline (T0), and 1 (T1) and 3 months (T3) after treatment start. The primary end-point was to evaluate the effect of SEV on p-cresol levels. RESULTS: Compared to baseline (T0, 7.4 ± 2.7 mg/mL), p-cresol mean concentration was significantly reduced in SEV patients after one (- 2.06 mg/mL, 95% CI - 2.62 to - 1.50 mg/mL; p < 0.001) and 3 months of treatment (- 3.97 mg/mL, 95% CI - 4.53 to - 3.41 mg/mL; p < 0.001); no change of plasma p-cresol concentration was recorded in placebo-treated patients. Moreover, P and LDL values were reduced after 3 months of treatment by SEV but not placebo. CONCLUSIONS: In conclusion, our study represents the first evidence that SEV is effective in reducing p-cresol levels in CKD patients in conservative treatment, and confirms its beneficial effects on inflammation and lipid pattern.

Bisphenol A and Bisphenol S release in milk under household conditions from baby bottles marketed in Italy.

A simple and sensitive validated analytical method based on liquid chromatography coupled to tandem fluorescence (FD) and ultraviolet (UV) spectrophotometry was applied to monitor the presence of bisphenol A and bisphenol S in plastic baby bottles marketed in Italy. The limits of detection (LOD) were 3.75 ng mL-1 and 80.00 ng mL-1, and those of quantification (LOQ) were 12.51 ng mL-1 and 260.00 ng mL-1 for BPA (FD detection) and for BPS (UV detection), respectively. BPA was found in only four samples, two samples undergone to microwave heating and two samples undergone to bottle warmer heating either at 40°C or at 80°C. Although the quantities of leached BPA were well below the reference dose for daily intake established by the European Food Safety Authority (EFSA) (4.0 µg kg-1 bw/day), the release of BPA and BPS from these plastic materials should be carefully considered by the government authorities to increase people's awareness on this issue and to protect the most vulnerable population group.

Effect of a Short-Course Treatment with Synbiotics on Plasma p-Cresol Concentration in Kidney Transplant Recipients.

OBJECTIVE: We evaluated whether a short-term course with synbiotics may lower plasma p-Cresol concentrations in kidney transplant patients (KTRs) who accumulate this uremic toxin both because of increased production by their dysbiotic gut microbiome and because of reduced elimination by the transplanted kidneys. METHODS: Thirty-six KTRs (29 males, mean age 49.6 ± 9.1 years) with transplant vintage > 12 months, stable graft function, and no episode of acute rejection or infection in the last 3 months were enrolled in this single-center, parallel-group, double-blinded, randomized (2:1 synbiotic to placebo) study. Synbiotic (Probinul Neutro, CadiGroup, Rome, Italy) or placebo was taken at home for 30 days, as 5 g powder packets dissolved in water three times a day far from meals. The main outcome measure was the decrease in total plasma p-Cresol measured by high-performance liquid chromatography at baseline and after 15 and 30 days of placebo or synbiotic treatment. RESULTS: After 15 and 30 days of treatment, plasma p-Cresol decreased by 40% and 33% from baseline (both p < 0.05), respectively, in the synbiotic group, whereas it remained stable in the placebo group. After 30 days of treatment, no significant change was observed in either group in renal function, glycemia, plasma lipids, or albumin concentration. Treatment was well tolerated and did not induce any change in stool characteristics. CONCLUSION: The results of this pilot study suggest that treatment with synbiotics may be effective to lower plasma p-Cresol concentrations in KTRs. Prospective larger scale, longer term studies are needed to establish whether cardiovascular prognosis could also be improved with this nutritional intervention.

Immobilized Artificial Membrane HPLC Derived Parameters vs PAMPA-BBB Data in Estimating in Situ Measured Blood-Brain Barrier Permeation of Drugs.

The affinity indexes for phospholipids (log kW(IAM)) for 42 compounds were measured by high performance liquid chromatography (HPLC) on two different phospholipid-based stationary phases (immobilized artificial membrane, IAM), i.e., IAM.PC.MG and IAM.PC.DD2. The polar/electrostatic interaction forces between analytes and membrane phospholipids (Δlog kW(IAM)) were calculated as the differences between the experimental values of log kW(IAM) and those expected for isolipophilic neutral compounds having polar surface area (PSA) = 0. The values of passage through a porcine brain lipid extract (PBLE) artificial membrane for 36 out of the 42 compounds considered, measured by the so-called PAMPA-BBB technique, were taken from the literature (P0(PAMPA-BBB)). The values of blood-brain barrier (BBB) passage measured in situ, P0(in situ), for 38 out of the 42 compounds considered, taken from the literature, represented the permeability of the neutral forms on "efflux minimized" rodent models. The present work was aimed at verifying the soundness of Δlog kW(IAM) at describing the potential of passage through the BBB as compared to data achieved by the PAMPA-BBB technique. In a first instance, the values of log P0(PAMPA-BBB) (32 data points) were found significantly related to the n-octanol lipophilicity values of the neutral forms (log P(N)) (r(2) = 0.782) whereas no significant relationship (r(2) = 0.246) was found with lipophilicity values of the mixtures of ionized and neutral forms existing at the experimental pH 7.4 (log D(7.4)) as well as with either log kW(IAM) or Δlog kW(IAM) values. log P0(PAMPA-BBB) related moderately to log P0(in situ) values (r(2) = 0.604). The latter did not relate with either n-octanol lipophilicity indexes (log P(N) and log D(7.4)) or phospholipid affinity indexes (log kW(IAM)). In contrast, significant inverse linear relationships were observed between log P0(in situ) (38 data points) and Δlog kW(IAM) values for all the compounds but ibuprofen and chlorpromazine, which behaved as moderate outliers (r(2) = 0.656 and r(2) = 0.757 for values achieved on IAM.PC.MG and IAM.PC.DD2, respectively). Since log P0(in situ) refer to the "intrinsic permeability" of the analytes regardless their ionization degree, no correction for ionization of Δlog kW(IAM) values was needed. Furthermore, log P0(in situ) were found roughly linearly related to log BB values (i.e., the logarithm of the ratio brain concentration/blood concentration measured in vivo) for all the analytes but those predominantly present at the experimental pH 7.4 as anions. These results suggest that, at least for the data set considered, Δlog kW(IAM) parameters are more effective than log P0(PAMPA-BBB) at predicting log P0(in situ) values for all the analytes. Furthermore, ionization appears to affect differently, and much more markedly, BBB passage of acids (yielding anions) than that of the other ionizable compounds.

Dioxin-like PCB levels in maternal and umbilical cord sera of people living near dump sites in southern Italy: a pilot study of biomonitoring.

The "Triangle of Death" is an area of southern Italy highly polluted due to the presence of numerous illegal waste sites. To assess whether environmental contamination could produce an increased intake of harmful chemicals, we determined the serum levels of the 12 polychlorinated biphenyls (PCBs) indicated as "dioxin like" by the World Health Organization (WHO) in both maternal blood serum and umbilical cord blood serum of people residing in that area. The study was performed on 32 pregnant women and cord serum of their 32 newborn babies. Maternal serum concentrations ranged from 0.019 to 10.426 ng/mL (mean 1.590 ± 2.002 ng/mL), while umbilical serum concentrations ranged from 0.018 to 16.980 ng/mL (mean 1.858 ± 3.558 ng/mL). To normalize the toxic impact of the different congeners, the levels were expressed as WHO-TEQ, i.e., toxic equivalent concentrations. Total PCB concentrations (ΣPCBs WHO-TEQ 1998) were 0.297 TEQ-ng/mL in maternal serum and 0.663 TEQ-ng/mL in cord blood. These levels are significantly higher not only than those found in subjects resident in other areas but also in comparison to those reported in other published studies. This suggests that the exposure of both pregnant mothers and their fetuses to environmental pollution could produce a high intake of PCBs. The study may be important for the public health authorities to evaluate the level of exposure to environmental pollutants in human residents in the Triangle of Death.

Into the toxicity potential of an array of parabens by biomimetic liquid chromatography, cell viability assessments and in silico predictions.

Five parabens (PBs) i.e., Methylparaben (MP), Ethylparaben (EP), Isopropylparaben (iPrP), Isobutylparaben (iBuP), Benzylparaben (BzP), and their parent compound i.e., para-hydroxy Benzoic Acid (pHBA), were studied both in vitro and in silico. Specifically, we determined their retention on several both protein- (Human Serum Albumin and α1-acidic glycoprotein) and (phospho) lipid- (immobilized artificial membrane (IAM)) based biomimetic stationary phases to evaluate their penetration potential through the biomembranes and their possible distribution in the body. The IAM phases were based either on phosphatidylcholine (PC) analogues i.e., PC.MG and PC.DD2 or on sphingomyelin (SPH). We also assessed their viability effect on breast cancer cells (MCF-7) via MTT assay subjecting the cells to five different PB concentrations i.e., 100 µM, 10 µM, 1 µM, 0.1 µM and 0.01 µM. Finally, their pharmacokinetics and toxicity were assessed by the ADMET Predictor™ software. Isopropylparaben was found to be more active than 17ß estradiol (E2) employed as positive control, on the screened cell line inducing cell proliferation up to 150 % more of untreated cells. Other analogues showed only a slight/moderate cell proliferation activity, with parabens having longer/branched side chain showing, on average, a higher proliferation rate. Significant linear direct relationships (for PC.DD2 r2 = 0.89, q2 = 0.86, for SPH r2 = 0.89, q2 = 0.85, for both P value < 0.05) were observed between the difference in proliferative effect between the readout and the control at 0.01 µM concentration and the retention on the IAM phases measured at pH 5.0 for all compounds but pHBA, which is the only analyte of the dataset supporting a carboxylic acid moiety. IAM affinity data measured at pH 7.0 were found to be related to the effective human jejunal permeability as predicted by the software ADMET® Predictor, which is relevant when PBs are added to pharmaceutical and food commodities.

New flurbiprofen derivatives: synthesis, membrane affinity and evaluation of in vitro effect on ß-amyloid levels.

Alzheimer's disease (AD) is characterized by irreversible and progressive loss of memory and cognition and profound neuronal loss. Current therapeutic strategies for the treatment of AD have been directed to a variety of targets with the aim of reversing or preventing the disease but, unfortunately, the available treatments often produce no significant clinical benefits. During the last decades compounds that inhibit or modulate γ-secretase, reducing ß amyloid (Aß) levels, have been considered as potential therapeutics for AD. Among these the (R)-enantiomer of flurbiprofen (FLU) seems to be very promising, but it shows low brain penetration. In this study, in order to improve the properties of FLU against Alzheimer's pathogenesis we synthesized some novel FLU lipophilic analogues. Lipophilicity of the new molecules has been characterized in terms of clogP, log K(C18/W) and log K(IAM/W) values. Permeability has been determined in both gastrointestinal PAMPA (PAMPA-GI) at different pH values and in brain blood barrier PAMPA (PAMPA-BBB) models. They were also tested for their ability to inhibit in vitro γ-secretase activity using rat CTXTNA2 astrocytes. Interestingly, the investigated molecules demonstrated to reduce Aß 42 levels without affecting the amyloid precursor protein APP level in a clear concentrations-dependent manner.

Lipophilicity profiling and cell viability assessment of a selected panel of endocrine disruptors.

Endocrine disruptors are chemicals widely used worldwide by industries in a variety of applications. Routinely exposure to these chemicals, even if at low doses, can cause damage effects on human health. In the present study, we evaluated toxic effects of nine chemicals, among which phthalates, using various cell lines to inspect their capability to interfere with cell proliferation and viability. Alongside, we investigated their affinity for phospholipids to assess the possible passage through biomembranes. Experimentally determined logkwIAM.MG values ranged from 1.37 to 3.49 whilst calculated log kwIAM.DD2 spanned from 1.80 to 5.21, supporting the target contaminants to exhibit lipophilicity moderate or very high. The achieved results were related to pharmacokinetic and toxicological properties by ADMET predictor™ and EPI Suite™ software. Triclosan and 4-Nonylphenol were found to be the most toxic against all cell lines screened, showing an IC50 of 30 µM for triclosan on human keratinocytes and of 50 µM for 4-Nonylphenol on human colorectal adenocarcinoma cells. Overall, even if the phthalates showed higher IC50 values (ranging from 170 µM to 280 µM), we can assert that all contaminants herein tested were able to interfere with cell growth and viability.

Exposure to Bisphenol A increases malignancy risk of thyroid nodules in overweight/obese patients.

Bisphenol A (BPA) is a widespread thyroid disruptor, but evidence about an association with thyroid cancer is weak. Excess body weight is a risk factor for thyroid cancer and affects activity of endocrine disruptors. Aim of the study was to investigate the association between BPA exposure and thyroid cancer, verifying the effect modification related to body weight. We performed a multicentre, cross-sectional study including consecutive patients referring for nodular goiter. The quantitative determination of BPA in serum samples was performed through high performance liquid chromatography system, coupled in tandem with ultraviolet and fluorescence detection. Ninety-six patients were included: 55 benign nodules, 41 thyroid cancers, 28 normal weight, and 68 overweight/obese. BPA was detected in 79 subjects. In the overall study population and in the group with BMI<25 kg/m2 BPA exposure was not significantly correlated to thyroid cancer (p = 0.08 and 0.759, respectively). In the group with BMI≥25 kg/m2, BPA-exposed subjects showed significantly higher risk of malignancy (OR: 5.3, p = 0.028). At multivariate analysis, such association was independent of smoking, alcohol consumption, occupational exposure, and phthalates exposure (p = 0.021 and 0.016, respectively), but was lost after adjustment for the presence of metabolic syndrome (p = 0.089). In overweight/obese subjects, BPA exposure was significantly associated with higher thyroid stimulating hormone levels. Our study suggests that BPA exposure is a risk factor for thyroid cancer in overweight/obese subjects.

Validation of an LC-MS/MS Method for the Determination of Abscisic Acid Concentration in a Real-World Setting.

One of the most relevant aspects in evaluating the impact of natural bioactive compounds on human health is the assessment of their bioavailability. In this regard, abscisic acid (ABA) has attracted particular interest as a plant-derived molecule mainly involved in the regulation of plant physiology. Remarkably, ABA was also found in mammals as an endogenous hormone involved in the upstream control of glucose homeostasis, as evidenced by its increase after glucose load. The present work focused on the development and validation of a method for the determination of ABA in biological samples through liquid-liquid extraction (LLE), followed by liquid mass spectrometry (LC-MS) of the extract. To test method suitability, this optimized and validated method was applied to a pilot study on eight healthy volunteers' serum levels to evaluate ABA concentration after consumption of a standardized test meal (STM) and the administration of an ABA-rich nutraceutical product. The results obtained could meet the demands of clinical laboratories to determine the response to a glucose-containing meal in terms of ABA concentration. Interestingly, the detection of this endogenous hormone in such a real-world setting could represent a useful tool to investigate the occurrence of impaired ABA release in dysglycemic individuals and to monitor its eventual improvement in response to chronic nutraceutical supplementation.

High-throughput and reliable assessments of the ionization constant of monoprotic organic acids through an arginine based mixed mode HPLC.

The charge state of a molecule is the single most prominent attribute ruling out its interactions with the surrounding environment. In a previous study, the retention of acids on the new Celeris™ Arginine (ARG) column was found to be predominantly driven by electrostatics and, specifically, their charge state. Therefore, we analysed 41 compounds in liquid chromatography with ultraviolet detection to study possible relationships between the analytical retention on this phase and the pKa of the acidic solutes. Highly significant relationships were observed indicating either a linear (r2 = 0.86) or a quadratic (r2= 0.89) trend. To improve the throughput of the method, this was transferred to LC mass spectrometry, allowing the analysis of a molecule every 3 mins. The developed method was found to be fast, reliable, accurate, easily automatable and simple to set up. Finally, the analytical column's being industrially manufactured and commercially available offers broad applicability.

Determination of dimethyl fumarate in desiccant and antimould sachets by reversed-phase liquid chromatography.

A method for the determination of dimethyl fumarate (DMF) in desiccant and antimould sachets, employed for protecting consumer products from humidity and mould, has been developed. The method is based on a solid-liquid extraction followed by HPLC-UV analysis. The method was validated with respect to recovery, linearity, limits of detection and quantitation and precision. The recovery was 98%. The correlation coefficient value (r) was equal to 0.94. Both intra- and inter-day precisions were studied at several concentration levels, being satisfactory in all cases (RSD < 5). Limits of detection and quantification values were in the low microgram per gram level, thus allowing the determination of DMF at concentrations below the limit established (0.1 mg/kg) by the recent EU Directive (Decision 2009/251/EC). The proposed procedure was applied for the determination of the target compound in 41 desiccant and antimould samples. DMF was detected in 39.0% of samples and its content in many samples exceeded the legal limits. The results of our analysis highlight the high risk of exposure to this powerful allergic sensitizer for consumers.