Commentary of the SKLM to the EFSA opinion on risk assessment of N-nitrosamines in food.

Dietary exposure to N-nitrosamines has recently been assessed by the European Food Safety Authority (EFSA) to result in margins of exposure that are conceived to indicate concern with respect to human health risk. However, evidence from more than half a century of international research shows that N-nitroso compounds (NOC) can also be formed endogenously. In this commentary of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG), the complex metabolic and physiological biokinetics network of nitrate, nitrite and reactive nitrogen species is discussed with emphasis on its influence on endogenous NOC formation. Pioneering approaches to monitor endogenous NOC have been based on steady-state levels of N-nitrosodimethylamine (NDMA) in human blood and on DNA adduct levels in blood cells. Further NOC have not been considered yet to a comparable extent, although their generation from endogenous or exogenous precursors is to be expected. The evidence available to date indicates that endogenous NDMA exposure could exceed dietary exposure by about 2-3 orders of magnitude. These findings require consolidation by refined toxicokinetics and DNA adduct monitoring data to achieve a credible and comprehensive human health risk assessment.

Distribution of nitrate/nitrite and toxic metals in the soil-potato system and its health risk assessment in Iran.

Potato is one of the essential food products whose health quality is greatly influenced by soil contamination and properties. In the current study, we have investigated the physicochemical characteristics of agricultural areas and the accumulation of nitrite/nitrate and metals in potato products in Hamedan, Iran. After determining the physicochemical characteristics of soil samples from four agricultural regions of Hamedan, 48 potato samples were collected from these regions. The heavy metals and nitrate/nitrite content were determined by ICP-OES and calorimetric methods, respectively. A negative correlation was observed between soil pH changes with nitrite/nitrate content and the accumulation of some heavy elements in potatoes. Furthermore, a positive correlation was found between soil phosphorus content and lead accumulation in potato. In present study, the amounts of lead, nitrate, and nitrite in 83.3%, 56%, and 12% of the collected samples were higher than the permissible limit reported by the World Health Organization (WHO), respectively. The EDI range for nitrate and nitrite was determined to be 130-260 and 1.4-2.7 µg/kg/day, respectively, which is much lower than the RfD set by the US Environmental Protection Agency (USEPA) for nitrite and nitrate. Among metal pollutants, the toxic risk caused by lead in potato consumers was higher than the threshold limit. In conclusion, our findings showed that the physicochemical characteristics of the soil could effectively increase the availability of metal pollutants and nitrite/nitrate to the potato product and significantly reduce its health quality. Therefore, monitoring these pollutants in the soil-potato system, preventing the entry of industrial wastewater, and managing the use of agricultural fertilizers can effectively improve the health of this product for consumers.

Plasma-activated liquid as a potential decontaminant in healthcare: assessment of antibacterial activity and use with cleaning cloths.

BACKGROUND: Cold air plasma (CAP) can generate plasma-activated liquids (PALs) with high concentrations of reactive oxygen (ROS) and nitrogen species (RNS), e.g., nitrites, with antimicrobial properties. AIM: We investigated the concentrations of ROS and RNS in saline PAL. We assessed planktonic bacterial inactivation by PAL and the decontamination of contaminated cleaning cloths. METHODS: Phosphate-buffered saline (PBS) was treated with an air-driven CAP jet for 90 or 300 s to generate PAL. The ROS and RNS were measured using quantitative fluorescent (2,7-dichlorofluorescin diacetate) and colourimetric (Greiss) assays. Isolates of MRSA and Escherichia coli were incubated in PAL overnight and inactivation measured through colony forming unit (cfu) assays. Sections of cleaning cloths were incubated with MRSA and E. coli, and treated with PAL for 1 h. Bacterial inactivation was measured through resazurin reduction assays. RESULTS: Nitrites increased from 0.1 µM in untreated PBS to 49.1 µM and to 94.0 µM in 90- and 300-s CAP-treated PAL, respectively. ROS increased from 30 µM in untreated PBS to 75 µM and to 103 µM in 90- and 300-s CAP-treated PAL, respectively. 90-s PAL reduced MRSA and E. coli viability (P<0.05) and 300-s PAL resulted in more than a 7-log reduction of both. One-hour treatment of contaminated cleaning cloths in PAL resulted in a 55% and 73% reduction in viable MRSA and E. coli, respectively (P<0.05). CONCLUSION: Inactivation of planktonic bacteria correlated with ROS and RNS concentrations. PAL reduced bacteria contaminated cleaning cloths. PAL has potential as a hospital disinfectant, including cleaning cloths.

Green Synthesis and Biosafety Assessment of MXene.

The rise of MXene-based materials with fascinating physical and chemical properties has attracted wide attention in the field of biomedicine, because it can be exploited to regulate a variety of biological processes. The biomedical applications of MXene are still in its infancy, nevertheless, the comprehensive evaluation of MXene's biosafety is desperately needed. In this review, the composition and the synthetic methods of MXene materials are first introduced from the view of biosafety. The evaluation of the interaction between MXene and cells, as well as the safety of different forms of MXene applied in vivo are then discussed. This review provides a basic understanding of MXene biosafety and may bring new inspirations to the future applications of MXene-based materials in biomedicine.

Wastewater treatment plant performance assessment using time-function-based effluent quality index and multiple regression models: the case of Bahir Dar textile factory.

Extensive water and chemicals are used in the textile industry processes. Therefore, treatment of textile wastewater is vital to protect the environment, maintain the public health, and recover resources. However, due to poor operation and plant performance the partially treated textile wastewater was directly discharged to a nearby river. Thus, the aim of this study was to characterize the wastewater physicochemical properties and evaluate the performance of the textile factory-activated sludge process wastewater treatment plant (WWTP) in Bahir Dar, Ethiopia. In inlet and outlet of the WWTP, samples were collected for 6 months and analyzed on-site and in a laboratory for parameters including, dissolved oxygen, pH, temperature, total Kjeldhal nitrogen (TKN), chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total suspended solids (TSS), total nitrogen (TN), total phosphorous (TP), nitrite, nitrate, and metallic compounds. The TSS, BOD5, COD, TP, nitrite, ammonia, and total chromium result were above the discharge limit with 73.2 mg/L, 48.45 mg/L, 144.08 mg/L, 7.9 mg/L, 1.36 mg/L, 1.96 mg/L, and 0.16 mg/L, respectively. Multiple regression models were developed for each overall, net moving average, and instantaneous effluent quality index (EQI). The predictor parameters BOD5, TN, COD, TSS, and TP (R2 = 0.995 to 1.000) estimated the net pollution loads of all predictors as 492.55 kg/day and 655.44 kg/day. Except TN, TKN, and NO3, the remaining six performance parameters were violating the permissible limit daily. Furthermore, the overall plant efficiency was predicted as 38 % and 42 % for the moving average and instantaneous EQI, respectively. Our study concluded that the integrated regression models and EQI can easily estimate the plant efficiency and daily possible pollution load.

The effects of inorganic nitrate supplementation on exercise economy and endurance capacity across the menstrual cycle.

Oral inorganic nitrate (NO3-) supplementation has been shown to increase bioavailable NO and provide potential ergogenic benefits in males; however, data in females is scarce. Estrogen is known to increase endogenous NO bioavailability and to fluctuate throughout the menstrual cycle (MC), being lowest in the early follicular (EF) phase and highest during the late follicular (LF) phase. This study examined the effects of oral NO3- supplementation on exercise economy, endurance capacity, and vascular health in young females across the MC. Ten normally menstruating females' MCs were tested in a double-blinded, randomized design during both the EF and LF phases of the MC. Participants consumed ∼13 mmol NO3-, in the form of 140 mL beetroot juice (BRJ) or an identical NO3--depleted placebo (PL) for ∼3 days before lab visits and 2 h before testing on lab visits. Plasma nitrate, nitrite, and estradiol were assessed, as was blood pressure and pulse wave velocity. Moderate-intensity exercise economy and severe intensity time to exhaustion (TTE) were tested on a cycle ergometer. As expected, plasma estradiol was elevated in the LF phase, and plasma nitrite and nitrate were elevated in the BRJ condition. Exercise economy was unaltered by BRJ or the MC, however TTE was significantly worsened by 48 s (∼10%) after BRJ supplementation (P = 0.04), but was not different across the MC with no interaction effects. In conclusion, NO3- supplementation did not affect exercise economy or vascular health and worsened aerobic endurance capacity (TTE), suggesting healthy females should proceed with caution when considering supplementation with BRJ.NEW & NOTEWORTHY Although inorganic nitrate (NO3-) supplementation has increased in popularity as a means of improving exercise performance, data in females at different phases of the menstrual cycle are lacking despite known interactions of estrogen with NO. This study revealed neither NO3- supplementation nor the menstrual cycle influenced exercise economy or vascular health in healthy young naturally menstruating females, while NO3- supplementation significantly worsened endurance capacity (10%) independent of the menstrual cycle phase.

Assessment of dietary exposure to food additives used in Polish processed meat products.

Dietary exposure assessments have been performed for nitrites, phosphates, erythorbic acid, and sodium erythorbate in processed meat in Poland. The exposure has been estimated based on the maximum level of use of additives contained in Regulation - tier 2 and the concentration of additives in processed meat - tier 3, additionally for nitrites. Daily intake was estimated using 24-h recall, repeated three times. Exposure analyses were correlated with the frequency of occurrence of food additives based on label analysis (tiers 2a and 3a). The mean nitrite intake from processed meat at tier 2 was 0.1 mg/kg bw (143% ADI), 0.08 mg/kg bw (118% - ADI) at tier 2a, 0.03 mg/kg bw (43% - ADI) at tier 3, and 0.026 mg/kg bw (37% - ADI) at tier 3a. The mean intakes of phosphate and erythorbic acid/sodium erythorbate from processed meat were 3.26 and 0.54 mg/kg bw (8.2% and 9% - ADI), respectively at tier 2. None of the respondents exceeded the ADIs for phosphates or erythorbic acid/sodium erythorbate. In contrast, nitrite consumption is of great concern because of significant ADI exceedances, particularly among young children.

Model-based assessment of mainstream nitrate/nitrite-dependent anaerobic methane oxidation and Anammox process in granular sludge at low temperature.

The process of nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) coupled with anaerobic ammonium oxidation (Anammox) is one of groundbreaking discoveries for nitrogen removal and methane emission reduction from wastewater simultaneously. Yet its treatment of mainstream wastewater at low temperature is still a major challenge. In this work, a one-dimensional granular sludge model incorporating Arrhenius conversion for temperature effects was constructed to depict the relationships among n-DAMO microorganisms and Anammox. The model framework was successfully evaluated with 380 days measurement data from a membrane granular sludge reactor (MGSR) operated at temperature of 20-10 °C and fed with ammonium and nitrite. The model could satisfactorily predict the kinetics of nitrogen removal rates, effluent nitrogen concentrations and biomass fractions in MGSR at varying temperatures. Despite the decrease in microbial activity of functional microorganisms, the coupled n-DAMO and Anammox process based on granule system in mainstream wastewater treatment achieved a TN removal efficiency of about 98 % and a stable nitrogen removal rate of 0.55 g L-1 d-1. The model developed is expected to facilitate fundamentally understanding the underlying mechanisms of the coupled process and provide proposals for its practical engineering application in wastewater treatment plants.

Occurrence of N-nitrosodimethylamine in roasted Alaska pollock fillets during processing and storage and preliminary cancer risk assessment.

BACKGROUND: Dried and salt-fermented fish products are important sources of N-nitrosodimethylamine (NDMA) exposure for human. As a potent carcinogen, NDMA was frequently detected in roasted Alaska pollock fillet products (RPFs), which is among the most common fish products in China. Until now, the occurrence and development of NDMA and its precursors (nitrites, nitrates and dimethylamine) in RPFs during processing and storage were not well elucidated, and safety evaluation of this fish product is also urgently needed. RESULTS: The presence of precursors in the raw material was verified and significant increase of nitrates and nitrites during processing was observed. NDMA was found generated during pre-drying (3.7 µg kg-1 dry basis) and roasting (14.6 µg kg-1 dry basis) process. Continuous increase in NDMA content can also be found during storage, especially at higher storage temperature. The 95th percentile of Monte Carlo simulated cancer risk (3.73 × 10-5 ) surpassed the WHO threshold (1.00 × 10-5 ) and sensitivity analysis implies the risk was mainly attributable to NDMA level in RPFs. CONCLUSION: The occurrence of NDMA in RFPs was mainly a result of endogenous factors originating in Alaska pollock during processing and storage rather than exogenous contamination, and temperature played a pivotal role. The preliminary risk assessment results suggest that long-term consumption of RPFs would impose potential health risks for consumers. © 2023 Society of Chemical Industry.

Determination of nitrate and nitrite levels in enteral nutrition formula marketed in Turkey: A risk assessment study.

In this study, the nitrite and nitrate levels of 37 enteral nutrition formulas of three different international brands marketed in Turkey were evaluated using high-performance liquid chromatography with diode-array detection (HPLC-DAD). Also, risk assessment of non-carcinogenic was calculated by models including hazard quotient (HQ) and hazard index (HI) in the deterministic model. Enteral nutrition formula consumption amounts obtained from individuals between the ages of 6-36 volunteers voluntarily participated in the study, and health risk assessments were calculated. The range of concentration of nitrate levels in enteral formulas of brands (B1, B2, and B3) was determined as 11.08 ± 2.88 (2.90-15.79), 11.64 ± 3.39 (2.92-22.93), and 10.66 ± 3.46 (4.92-15.37) mg/kg, respectively. The range of concentration of nitrite levels in enteral formulas of brands (B1, B2, and B3) was determined as 4.18 ± 1.10 (1.86-5.82), 3.70 ± 1.25 (1.29-5.26), and 3.38 ± 1.67 (2.00-5.29) mg/kg, respectively. The mean nitrate and nitrite levels of exposure from consumption of enteral nutrition formulas were determined as 0.14 and 0.11, 0.06 and 0.07 mg/kg bw/day for females and males, respectively. Nitrate and nitrite exposure levels calculated in the study did not exceed the ADI threshold values given by JECFA. The average HQ value calculated for the exposure of nitrate to male and female individuals was less than 1. Still, the P95 values calculated for nitrate were greater than 1.00 except for female and male individuals (24-36 ages). The HI value was found to be greater than 1.00 in all age groups of both genders. The presence of nitrites and nitrates in enteral nutrition formulas may lead to health problems in sensitive groups.

The Clinical Assessment and Treatment of Inhalant Abuse.

Although inhalant abuse is common, it is one of the most neglected and overlooked forms of substance abuse. Inhalants refer to a wide variety of substances including volatile solvents, aerosols, gases, and nitrites. The mechanism of action of inhalants has not been fully defined. Several molecular targets contribute to the pharmacology, including ion-channel proteins that control neuronal excitability. These agents interact with various receptors and can cause changes in cell-membrane fluidity and nerve-membrane ion channels. Three main pharmacologic categories of inhalants, namely, volatile solvents and anesthetic gases, nitrous oxide, and volatile alkyl nitrites, have distinct pharmacologies, mechanisms of action, and toxicities. Inhalants are linked to multisystem damage affecting the pulmonary, cardiac, dermatologic, renal, hematologic, gastrointestinal, hepatic, and neurologic systems. Chronic inhalant abuse can also cause psychiatric, cognitive, behavioral, and anatomical deficits in humans, leading to reduced productivity and quality of life. Inhalant abuse during pregnancy is associated with fetal abnormalities. Clinical assessment for inhalant abuse should be done systematically. After decontamination and stabilization of the patient, further history and physical examination is necessary to establish an appropriate diagnosis based on the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition. Laboratory testing for inhalant abuse is very limited, and imaging studies may be helpful in certain situations. The treatment of inhalant use disorder is similar to that of other substance abuse disorders and includes supportive care, pharmacotherapy, and behavioral therapy. Preventive measures are essential.

Application of oleaster leaves (Elaeagnus angustifolia L.) essential oil and natural nanoparticle preservatives in frankfurter-type sausages: An assessment of quality attributes and stability during refrigerated storage.

The effects of oleaster leave essential oil (OLEOs: 1000 and 2000 ppm) in combination with nisin nanoparticles (200 ppm) and ε-polylysine nanoparticles (2000 ppm) on the physicochemical, microbiological and sensory properties of the emulsion-type sausages without added chemical nitrite/nitrate salts were evaluated during 45 days of storage. Nanoparticle attributes were assessed, including encapsulation efficiency (EE%), zeta potential, nanoparticles size, FTIR analysis, and thermal stability (DSC). Overall, ε-PL nanoparticles (ε-PL-NPs) were thermally more stable and showed higher EE% (91.52%) and zeta potential (37.80%) as compared to nisin nanoparticles (82.85%) and (33.60%), respectively. The use of combined ε-PL-NPs (2000 ppm) + Ni-NPs (200 ppm) with oleaster leaves essential oil (2000 ppm) resulted in a higher pH value (5.88), total phenolic content (10.45 mg/100 g) and lower TBARS (2.11 mg/kg), and also decreased total viable bacteria (1.28 Log CFU/g), Clostridium perfringens (1.43 Log CFU/g), E. coli (0.24 Log CFU/g), Staphylococcus aureus (0.63 Log CFU/g), and molds and yeasts (0.86 Log CFU/g) count in samples at day 45 in comparison to the control (120 ppm nitrite). The consumers approved sensory traits in nitrite-free formulated sausages containing ε-PL-NPs and Ni-NPs combined with OLEOs.

Comparative assessment of energy generation from ammonia oxidation by different functional bacterial communities.

Bioelectrochemical ammonia oxidation (BEAO) in a microbial fuel cell (MFC) is a recently discovered process that has the potential to reduce energy consumption in wastewater treatment. However, level of energy and limiting factors of this process in different microbial groups are not fully understood. This study comparatively investigated the BEAO in wastewater treatment by MFCs enriched with different functional groups of bacteria (confirmed by 16S rRNA gene sequencing): electroactive bacteria (EAB), ammonia oxidizing bacteria (AOB), and anammox bacteria (AnAOB). Ammonia oxidation rates of 0.066, 0.083 and 0.082 g NH4+-N L-1 d-1 were achieved by biofilms enriched with EAB, AOB, and AnAOB, respectively. With influent 444 ± 65 mg NH4+-N d-1, nitrite accumulation between 84 and 105 mg N d-1 was observed independently of the biofilm type. The AnAOB-enriched biofilm released electrons at higher potential energy levels (anode potential of 0.253 V vs. SHE) but had high internal resistance (Rint) of 299 Ω, which limits its power density (0.2 W m-3). For AnAOB enriched biofilm, accumulation of nitrite was a limiting factor for power output by allowing conventional anammox activity without current generation. AOB enriched biofilm had Rint of 18 ± 1 Ω and yielded power density of up to 1.4 W m-3. The activity of the AOB-enriched biofilm was not dependent on the accumulation of dissolved oxygen and achieved 1.5 fold higher coulombic efficiency when sulfate was not available. The EAB-enriched biofilm adapted to oxidize ammonia without organic carbon, with Rint of 19 ± 1 Ω and achieved the highest power density of 11 W m-3. Based on lab-scale experiments (scaling-up factors not considered) energy savings of up to 7 % (AnAOB), 44 % (AOB) and 475 % (EAB) (positive energy balance), compared to conventional nitrification, are projected from the applications of BEAO in wastewater treatment plants.

Risk Assessment of Nitrite and Nitrate Intake from Processed Meat Products: Results from the Hellenic National Nutrition and Health Survey (HNNHS).

Long-term exposure to a high nitrite and nitrate intake through processed meat is of concern, as it has been related to adverse health effects. Individual consumption data from 2152 participants (46.7% males) in the Hellenic National Nutrition and Health Survey (HNNHS) were linked with current Maximum Permitted Levels (MPLs) to calculate exposure to nitrite and nitrate from processed meat products (assessed as nitrite equivalent), evaluate potential risk and identify the major contributors. Processed meat intakes were determined by combining data from 24 h recalls and frequency of consumption reported in Food Propensity Questionnaires (FPQs). Median exposure was estimated to be within safe levels for all population groups. However, 6.6% (n = 143) of the consumers exceeded the Acceptable Daily Intake (ADI) of nitrite (0.07 mg/kg bw/day), of which 20.3% were children aged 0-9 years (N = 29) (15.3% of all children participants in the study, N = 190). In total, pork meat was the major contributor (41.5%), followed by turkey meat (32.7%) and sausages (23.8%), although contribution variations were found among age groups. The outcomes are of public health concern, especially exposure among children, and future research is warranted to evaluate possible associations with health effects, by using more refined occurrence data if available.

Risk control and assessment of sulfide-rich sediment remediation by controlled-release calcium nitrate.

Nitrate stimulation is widely used in sediment remediation to eliminate sulfides, degrade organic pollutants and immobilize phosphorus. However, the environmental risks of nitrate escape and the subsequent release of pollutants (e.g. nitrite, ammonium and trace metals) to water bodies during its application has received less attention. In this study, controlled-release nitrate pellets (SedCaN pellets) were manufactured and applied at different sediment depths to examine their effectiveness in controlling the risk of nitrate escape and subsequent pollutant release. The germination of submerged plant was also analyzed to assess the ecological risks associated with the remediated sediment. The results showed that the SedCaN pellets slowly released calcium nitrate, which led to denitrifying sulfide oxidation, organic matter degradation and the immobilization of phosphorus as a calcium-bound species. Gas production by denitrification increased the sediment porosity (0.3-2.2%) and led to the concomitant release of nitrite, ammonium, and heavy metals, creating secondary risks. Application of the SedCaN pellets at depth decreased the nitrate escape and the secondary risks, presumably by means of a capping effect of the upper sediment. The release of nitrate, ammonium, Ni and Cu were partially limited by 91.6%, 19.0%, 61.6% and 57.4% when SedCaN pellets were incorporated into deeper sediments (7-9 cm). Moreover, the range of sulfide oxidation extended to the upper and lower sediments in the profile (column), while the sulfide oxidation efficiency reached 85.9-95.0%. Finally, increased germination of Bacopa monnieri (20.0-26%) demonstrated that in comparison to reference materials the ecological risks of the treated sediments was reduced and the habitat function of sediment was restored after nitrate-stimulating remediation. The results of this study provide valuable guidelines for nitrate-stimulating remediation of sulfide-rich (black-odor) sediments.

Chemometric and risk assessment of nitrogen composition of atmospheric rainwater from diverse surfaces in Rivers State, Nigeria.

Organic and inorganic nitrogen ions in the environment play important role across environmental matrices. Rainwater samples collected from ambient and different roofing surfaces (zinc, aluminium, asbestos and stone-coated roofing sheets) from selected locations at Ogale, Rumuodomaya/Rumuodome, Diobu and Chokocho within Rivers State, Niger Delta, Nigeria, from April to June, July to August and September to October depicting three regiments of early, mid and late rains. The samples were analysed for Kjeldahl nitrogen, ammonium, nitrate and nitrite using APHA methodology. Quantitative assessment showed that Kjeldahl nitrogen were in range of 0.11 to 28.05 mg/L; ammonium 0.50 to 20.22 mg/L, nitrate from 0.12 to 22.69 mg/L and nitrite from 0.15 to 3.90 mg/L. Parameters decreased from early to late rain, which can be attributed to rain dilution factor potential, wind pattern and emission from anthropogenic sources that influenced the rainwater quality across surfaces. Nitrogen results showed that dry and wet deposition has great impact; atmospheric aerosols and biogeochemical interactions can affect water quality. Monthly variation showed that Ogale had high regression compared to other locations due to close proximity to oil and gas emission and marine contribution. Neutralization factor showed that nitrate-nitrite compounds have strong correlation with ammonium ion. Non-carcinogenic risk assessment using US EPA model showed hazard index less than one (1), thus no associated health effect of nitrate and nitrite in rainwater. In conclusion, it is evident that nitrate/nitrite levels and other nitrogen derivatives in rainwater in crude oil-producing Niger Delta and its continuous consumption can cause negative health outcome.

Determination of the geographic origin of 52 honey samples based on the assessment of anionic content profiling with a new algorithm using monolithic column-based micellar nano-liquid chromatography.

In the present study, a new micellar nano LC-UV was, for the first time, reported for the separation and determination of five anions (chloride, nitrite, bromide, sulfate and nitrate) in 52 honey samples. Based on this approach, a graphene oxide-based monolithic column was prepared and applied for the samples. Various amounts of hexadecyltrimethyl-ammonium bromide (HTAB) in the mobile phase were used in order to optimize the separation conditions. The baseline separation was achieved using mobile phase with 25/75% (v/v) ACN/10 mM phosphate buffer at pH 3.4, while the amount of HTAB was optimized as 0.22 mM in the mobile phase. The whole method was validated and it leads to high sensitivity. The LOD values were found in the range of 0.02-0.22 µg/kg, while LOQ values were found in the range of 0.06-0.18 µg/kg. The method allowed to achieve sensitivity analyses of anionic content in 52 honey samples. All data were evaluated using a new algorithm for geographic origin discrimination. K-nearest neighbor algorithm (K-NN), cubic support vector classifier (K-DVS), and K-Mean cluster analysis were used for geographic origin discrimination of honeys. The accuracy of the whole model was calculated as 94.4% with the K-DVS method. The samples from five provinces were classified 100% correctly, while two of them were classified with one misclassification, with an accuracy of 89.9% and 83.3%, respectively. PRACTICAL APPLICATION: The new platforms and advanced technologies are crucial for advanced food analysis. In this article, a novel methodology was attempted for the determination of geographic origin of 52 honey samples. In this sense, micellar nano LC technique with a homemade monolithic nano-column was, for the first time, applied for the anion analysis using a new algorithm.

A Model Assessment of the Occurrence and Reactivity of the Nitrating/Nitrosating Agent Nitrogen Dioxide (•NO2) in Sunlit Natural Waters.

Nitrogen dioxide (•NO2) is produced in sunlit natural surface waters by the direct photolysis of nitrate, together with •OH, and upon the oxidation of nitrite by •OH itself. •NO2 is mainly scavenged by dissolved organic matter, and here, it is shown that •NO2 levels in sunlit surface waters are enhanced by high concentrations of nitrate and nitrite, and depressed by high values of the dissolved organic carbon. The dimer of nitrogen dioxide (N2O4) is also formed in the pathway of •NO2 hydrolysis, but with a very low concentration, i.e., several orders of magnitude below •NO2, and even below •OH. Therefore, at most, N2O4 would only be involved in the transformation (nitration/nitrosation) of electron-poor compounds, which would not react with •NO2. Although it is known that nitrite oxidation by CO3•- in high-alkalinity surface waters gives a minor-to-negligible contribution to •NO2 formation, it is shown here that NO2- oxidation by Br2•- can be a significant source of •NO2 in saline waters (saltwater, brackish waters, seawater, and brines), which offsets the scavenging of •OH by bromide. As an example, the anti-oxidant tripeptide glutathione undergoes nitrosation by •NO2 preferentially in saltwater, thanks to the inhibition of the degradation of glutathione itself by •OH, which is scavenged by bromide in saltwater. The enhancement of •NO2 reactions in saltwater could explain the literature findings, that several phenolic nitroderivatives are formed in shallow (i.e., thoroughly sunlit) and brackish lagoons in the Rhône river delta (S. France), and that the laboratory irradiation of phenol-spiked seawater yields nitrophenols in a significant amount.

Juvenile exposure and adult risk assessment with single versus repeated exposure of melphalan in the germ cells of male SD rat: Deciphering the molecular mechanisms.

Melphalan significantly contributes to the increase in childhood cancer survival rate. It acts as a gonadotoxic agent and leads to testes damage, dysbalance in gonadal hormones, and impairment in the germ cell proliferation. Therefore, it might be a potent threat to male fertility in individuals who have undergone melphalan treatment during childhood cancer. However, the molecular mechanisms of melphalan-induced gonadal damage are not yet fully explored and they need to be investigated to determine the benefit-risk profile. In the present study, juvenile male SD rats were subjected to single and intermittent cycles of melphalan exposure in a dose-dependent (0.375, 0.75 and 1.5 mg/kg) manner. Methods of end-points evaluations were quantification of micronuclei formation in peripheral blood, sperm count, sperm motility and head morphology, sperm and testicular DNA damage, histological studies in testes, oxidative/nitrosative stress parameters. A single cycle of exposure at high dose (1.5 mg/kg) produced significant effect on micronuclei formation only after the first week of exposure, whereas failed to produce significant effect at the end of the sixth week. Intermittent cycles of exposure at the dose of 1.5 mg/kg produced significant alterations in all the parameters (micronuclei in peripheral blood, testes and epididymides weight and length, MDA, GSH and nitrite levels, sperm count and motility, sperm head morphology, testicular and sperm DNA damage, protein expression in testes and histological parameters). So, time of exposure as well as the amount of exposure (total dosage administered) is critical in determining the magnitude of the damage in germ cell risk assessment.

Nitrites as precursors of N-nitrosation in pharmaceutical samples - A trace level analysis.

The ultra-performance liquid chromatography (UPLC) method, which involves pre-column derivatization of nitrite with 2,3-diaminonaphthalene (DAN) to form 2,3-naphthotriazole (NAT), offers the advantages of easy sample preparation, simple derivatization, stable derivatives, rapid analysis, high sensitivity and specificity and lack of interferences for determining nitrite in pharmaceutical samples. Determination of NAT was performed on a an Acquity UPLC HSS T3 column using a gradient elution of 0.1% formic acid with acetonitrile at flow rate of 0.4 mL/min and temperature at 45 °C. The single-quadrupole mass detector was operated in the positive ion mode. Quadrupole mass analyser was employed in selected ion monitoring mode using a target ion at m/z = 170 as [M+H]+. The UPLC-MS method was validated as per International Council on Harmonization (ICH) guidelines in terms of linearity, limit of detection, limit of quantification, selectivity, accuracy, precision, intermediate precision and stability. The UPLC-MS method was demonstrated to be applicable for the determination of nitrite in various pharmaceutical samples. The proposed UPLC-MS method was used to study the effect of nitrite content in pharmaceutical products on the formation of N-nitrosamines. The high importance of nitrites in relation to the N-nitrosation reaction was discussed. As deduced from theory and justified by the presented results, reducing the nitrite concentration could definitely solve the N-nitrosamine contamination. Nitrites, unlike secondary and tertiary amines, are universal precursors to any N-nitrosamine, so this solution is easily transferable to any relevant pharmaceutical product.