Determination of Cotinine, 3'-Hydroxycotinine and Nicotine 1'-Oxide in Urine of Passive and Active Young Smokers by LC-Orbitrap-MS/MS Technique.

Tobacco smoke is probably the most significant factor conducing to toxic xenobiotics exposure to humans. The aim of the study was to develop a rapid and sensitive method for the determination of selected nicotine metabolites in urine of tobacco smokers and passive smokers. The method for removing protein and extracting the metabolites involved the centrifugation of urine with acetonitrile. Cotinine, trans-3'-hydroxycotinine, and (2'S)-nicotine 1'-oxide in the supernatant were determined using the LC-Orbitrap-MS/MS technique, with the selected ion monitoring (SIM) and parallel reaction monitoring (PRM) modes used. The recovery of these analytes added to the urine samples ranged from 72% to 101%. Repeatability and reproducibility were less than 3.1% and 10.1%, respectively. The study was carried out among medical students. The group was selected as representatives of young people and who as future physicians should be more aware of the effects of nicotine use. Concentration levels of cotinine and trans-3'-hydroxycotinine determined in ng/mL in the urine of cigarette smokers were 70- and 58-fold higher, respectively, compared to passive smokers. Higher concentrations were recorded in the urine of those passively exposed to tobacco smoke than in non-smokers, confirming that passive exposure to tobacco smoke is not harmless to the human body. However, no significant differences were observed in the concentration of (1'S,2'S)-nicotine 1'-oxide in the samples of individuals from various groups.

Women suffering from systemic lupus erythematosus are characterized by low blood levels of α-dicarbonyl compounds.

Introduction: Systemic lupus erythematosus (SLE) is a chronic, autoimmune disease, often characterised by severe course and unclear etiopathogenesis. The reaction of protein glycoxidation, also known as glycation, may be linked to etiopathogenesis of SLE. Advanced glycation end-products (AGEs) exhibit cytotoxic properties, affect cellular signalling, impair functions of extracellular proteins, and may act as neoepitopes. Glucosone (GS), glyoxal (GO), and methylglyoxal (MGO) are examples of α-dicarbonyl compounds (α-DCs) partaking in glycoxidation. The study aimed to evaluate concentrations of these three compounds in blood serum of SLE patients, and to compare the results with healthy individuals. Material and methods: 31 women suffering from SLE and 26 healthy individuals were included in the study. High-performance liquid chromatography with fluorescence detection was applied to evaluate concentrations of α-DCs in their serum samples. Correlations between the results and parameters such as disease duration time, age, glomerular filtration rate (GFR), Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K), and creatinine were analysed. Results: The SLE patients exhibited lower concentrations of glucosone, glyoxal, and methylglyoxal than the control group. Analysis of correlations showed a difference between the examined groups. Conclusions: In women suffering from SLE the course of α-DCs metabolism is altered. SLE patients are characterised by low serum levels of α-DCs. We hypothesise that either hindered proteasomal degradation or fast consumption of α-DCs in oxidative conditions may cause the observed low concentration of these compounds.

Polycyclic Aromatic Hydrocarbons (PAHs) in Grilled Marshmallows.

The aim of this study was to assess potential health risks among children and adolescents consuming various grilled marshmallows using a survey and to determine polycyclic aromatic hydrocarbons (PAHs) in these food products. PAH analysis in grilled marshmallows included a dilution stage with deionized water and liquid-liquid extraction with cyclohexane and solid-phase extraction (SPE). PAH fractions were initially analyzed via high-performance thin-layer chromatography, and PAH concentrations were determined via gas chromatography with a tandem mass detector using the selective reaction monitoring (SRM) mode. This study on the consumption of grilled marshmallows was conducted among approximately 300 children and adolescents. The preliminary results indicated that "raw" marshmallows did not contain PAHs. However, the obtained data suggested the exposure of young people to carcinogenic PAHs from grilled marshmallows (63.5% of them consumed marshmallows). Carcinogenic benzo(a)pyrene (BaP) was determined in all samples. The profile of PAH concentrations in the extracts isolated from various grilled types of marshmallows was similar (r2 > 0.8000), regardless of the grilling method. Compared to the white sugar confection, higher concentrations of PAHs were determined in multicolored marshmallows. The lack of social awareness about exposure to carcinogenic substances is alarming.

The Risk of Oral Cancer and the High Consumption of Thermally Processed Meat Containing Mutagenic and Carcinogenic Compounds.

The International Agency for Research on Cancer has classified the consumption of heat-processed meat as a direct human carcinogen and the consumption of red meat as a probable carcinogen. Mutagenic and carcinogenic compounds present in meat dishes include, among others, polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatic amines (HAAs). These compounds can cause the development of gastrointestinal cancer. Oral cancer is one of the world's research priorities due to the ever-increasing incidence rate. However, the effect of diet on oral cancer is still a poorly recognized issue. The aim of this study was to assess the relationship between the risk of oral cancer and dietary ingredients with a particular emphasis on red meat and thermally processed meat. This study was conducted among patients with oral cancer in 2022 and 2023. The shortened standardized Food Frequency Questionnaire (FFQ) and a multivariate regression statistical analysis were used. The high consumption of red meat in general and thermally processed meat, especially smoked, fried, roasted and boiled, increases the risk of oral cavity cancer. Limiting the consumption of meat products and modifying the methods of preparing meat dishes may reduce exposure to carcinogenic compounds from the diet and thus reduce the risk of developing oral cancer.

Polycyclic Aromatic Hydrocarbons (PAHs) in Roasted Pork Meat and the Effect of Dried Fruits on PAH Content.

Diet is one of the main factors affecting human health. The frequent consumption of heat-treated meat has been classified as both directly carcinogenic to humans and as a risk factor, especially in the case of cancers of the gastrointestinal tract. Thermally processed meat may contain harmful muta- and carcinogenic compounds, including polycyclic aromatic hydrocarbons (PAHs). However, there are natural ways to reduce the risk of diet-related cancers by reducing the formation of PAHs in meat. The purpose of this study was to determine changes in PAH levels in pork loin dishes prepared by stuffing the meat with dried fruits (prunes, apricots and cranberries) and baking it in a roasting bag. High-performance liquid chromatography with fluorescence detection (HPLC-FLD) was used to conduct a quantitative analysis of seven PAHs. Recovery results ranged from 61 to 96%. The limit of detection (LOD) was 0.003 to 0.006 ng/g, and the limit of quantification (LOQ) was 0.01 to 0.02 ng/g. Gas chromatography-mass spectrometry (GC-MS/MS) was used to confirm the presence of PAHs in food. The total PAH content of the roasted pork loin was 7.4 ng/g. This concentration decreased by 35%, 48% and 58% when the meat was roasted with apricots, prunes and cranberries, respectively. The cranberries also inhibited the formation of benzo(a)pyrene to the greatest extent. Thermally treating meat stuffed with dry fruits may be a simple and effective way to prepare foods with reduced levels of mutagens and carcinogens belonging to PAHs, and thus reduce the risk of cancer.

Application of High-Performance Liquid Chromatography with Fluorescence Detection for Non-Polar Heterocyclic Aromatic Amines and Acridine Derivatives Determination in Pork Loin Roasted in a Roasting Bag.

Heat treatment of meat can lead to the formation of carcinogenic organic compounds. The influence of dried fruits on the formation of non-polar heterocyclic aromatic amines (carbolines) and nitrogen derivatives of polycyclic aromatic hydrocarbons (azaarenes) in roasted pork loin was elucidated. Two hundred grams of fruit per 1 kg of meat were used as stuffing. Carbolines, derivatives of pyridoimidazole and pyridoindole, and azaarenes (benzoacridines and dibenzoacridines) were determined by means of high-performance liquid chromatography with fluorescence detection. The total concentration of six δ-, γ- and α-carbolines in roasted pork loin was 1.3 ng/g. This content decreased by 64%, 58%, and 54% in pork loin stuffed with prunes, apricots, and cranberries, respectively. Concentrations of ß-carbolines (harmane and norharmane) increased under the influence of added fruits. The norharmane content increased the most, from 2.2 ng/g in the control sample to 12.3 ng/g in meat prepared with cranberries. The harmane content increased from 1.0 ng/g to 3.6 ng/g in meat with prunes. The total concentration of azaarenes (two benzoacridines and dibenzo[a,c]acridine), which was close to 0.1 ng/g, decreased in dishes with prunes and apricots by 54% and 12%, respectively. Azaarenes were not found in samples of meat stuffed with cranberries.

Consumption of Thermally Processed Meat Containing Carcinogenic Compounds (Polycyclic Aromatic Hydrocarbons and Heterocyclic Aromatic Amines) versus a Risk of Some Cancers in Humans and the Possibility of Reducing Their Formation by Natural Food Additives-A Literature Review.

(1) Background: Thermal treatment of high-protein food may lead to the formation of mutagenic and carcinogenic compounds, e.g., polycyclic aromatic hydrocarbons and heterocyclic aromatic amines. Frequent consumption of processed meat was classified by the International Agency for Research on Cancer as directly carcinogenic for humans. (2) Methods: A literature review was carried out based on a search of online databases for articles on consuming thermally processed meat containing carcinogenic compounds versus a risk of cancers in humans published between 2001 and 2021. (3) Results: A review of the current literature on the participation of PAHs and HAA in the formation of certain neoplasms indicates a positive relationship between diet and the incidences of many cancers, especially colon cancer. A simple way to obtain dishes with reduced contents of harmful compounds is the use of spices and vegetables as meat additives. These seasonings are usually rich in antioxidants that influence the mechanism of HAA and PAH synthesis in food. (4) Conclusions: As there is a growing risk of a cancer tendency because of exposing humans to PAHs and HAAs, it is extremely vital to find a simple way to limit carcinogenic compound synthesis in a processed proteinaceous food. Disseminating the knowledge about the conditions for preparing dishes with a reduced content of carcinogenic compounds could become a vital element of cancer prevention programs.

The effect of onion and garlic on non-polar heterocyclic aromatic amines (α-, ß-, γ- and δ-carbolines) formation in pan-fried meat and gravy.

Thermal treatment of protein-rich food can lead to the formation of biologically active heterocyclic aromatic amines (HAAs). One of the methods to learn how to reduce the content as well as the influence of these compounds on heath is the study of factors inhibiting their synthesis. In the current investigation, the effect of onion and garlic on the formation of six possibly carcinogenic non-polar HAAs (α-, γ- and δ-carbolines) and two co-mutagenic ß-carbolines (harmane and norharmane) was evaluated by comparing their contents in meat and gravy samples obtained from pan-fried pork dishes prepared in the presence and absence of these vegetables. Carbolines were isolated from food samples by solid phase extraction. The quantitative analysis was performed by high-performance liquid chromatography with fluorescence detection. The concentrations of individual compounds in dishes prepared without added vegetables ranged from 0.02 ng g-1 (3-amino-1,4-dimethyl-5 H-pyrido(4,3-b)indole; Trp-P-1) to 10.1 ng g-1 of meat (2-amino-9 H-pyrido[2,3-b]indole; AαC). Onion (30 g/100 g of meat) and garlic (15 g/100 g of meat) lowered the total content (in meat and gravy) of the α-, δ- and γ-carbolines in the range from 52% to 87%. In contrast, onion caused an increase in the norharmane concentration both in meat and gravy. The percentage of carbolines in the gravies (assuming that their total content in meat and gravy is 100%) was higher in dishes prepared with onion and garlic than in dishes without these seasonings.

Human exposure to biologically active heterocyclic aromatic amines arising from thermal processing of protein rich food

Frequent consumption of thermally processed meat was classified by the International Agency for Research on Cancer to directly carcinogenic agents for humans. During the heat treatment of high protein food, mutagenic and carcinogenic, as well as neurotoxic heterocyclic aromatic amines are formed. Epidemiological studies confirm that exposure to some of these compounds may increase the risk of cancer in humans, especially the colon cancer. Most heterocyclic amines contain fried and grilled meat products, and the lowest content of these compounds can be found in boiled and slightly baked dishes. The use of spices and vegetable additives with antioxidant properties allows to obtain dishes with reduced content of these xenobiotics. An effective way to reduce human exposure to cancerogenic amines may be simultaneous consumption, together with meat dishes, products containing fiber which can adsorb molecules of heterocyclic amines in the gastrointestinal tract, as well as enrichment of the diet in the crucifers plants, as isothiocyanates released from them can inhibit the metabolic activation processes of heterocyclic amines. Raising the public awareness of the formation of mutagenic and carcinogenic compounds, including heterocyclic aromatic amines, during the intensive heat treatment of high protein food, as well as the dissemination of knowledge on the conditions regarding the preparation of dishes with reduced content of such compounds could become one of the components of cancer prevention programs in Poland.

Influence of the toothpaste with brazilian ethanol extract propolis on the oral cavity health.

Propolis-based therapeutic agents represent this potential for the development of new drugs in dental care. The aim of a clinical-cohort study was to determine the influence of application of toothpaste enriched with Brazilian extract of propolis (EEP) on health status of oral cavity. Laboratory analysis was conducted in order to assess the chemical composition of EEP including total phenolic compounds, the DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity, ABTS radical cation scavenging activity, and FRAP assay. Clinical research involved two groups of subjects comprising 32 adult patients, with assessment based on the preliminary evaluation of the state of their marginal periodontium. The investigation of oral health indices API, OHI, and SBI and microbiological examination of oral microflora were also carried out. Results obtained indicated time-dependent microbial action of EEP at 50 mg/L concentration, with antimicrobial activity against Gram-positive bacteria. The total decrease of API, OHI, and SBI mean values was observed. Hygienic preparations with 3% content of Brazilian ethanol extract of green propolis (EEP) efficiently support removal of dental plaque and improve the state of marginal periodontium.

Ethanolic extract of Brazilian green propolis sensitizes prostate cancer cells to TRAIL-induced apoptosis.

Prostate cancer represents an ideal disease for chemopreventive intervention. Propolis possesses immuno-modulatory, anti-tumour and chemopreventive properties. The tumour necrosis factor-related apoptosis-inducing ligand (TRAIL) is an important endogenous anti-cancer agent that induces apoptosis selectively in tumour cells. However, some cancer cells are resistant to TRAIL-mediated apoptosis. Naturally occurring phenolic and polyphenolic compounds sensitize TRAIL-resistant cancer cells and augment the apoptotic activity of TRAIL. The ethanolic extract of Brazilian green propolis (EEP) is rich in phenolic components. Our in vitro results indicate the potential targets in the TRAIL-induced apoptotic pathway for the cancer chemopreventive activity of Brazilian propolis. We examined the cytotoxic and apoptotic effects of Brazilian EEP and its bioactive components in combination with TRAIL on LNCaP prostate cancer cells. The chemical composition of Brazilian green propolis was determined by high performance liquid chromatography-diode array detection. The cytotoxicity was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl-tetrazolium and lactate dehydrogenase assays. Apoptosis was detected using annexin V-FITC by flow cytometry and fluorescence microscopy. The mitochondrial membrane potential (∆Ψm) was evaluated using DePsipher staining by fluorescence microscopy. Flow cytometry was used to analyse death receptor (TRAIL-R1 and TRAIL-R2) expression in LNCaP cells. The inhibition of nuclear factor-κB (NF-κB) (p65) activation in cancer cells was confirmed by the ELISA-based TransAM NF-κB kit. The LNCaP cells were shown to be resistant to TRAIL-induced apoptosis. Our study demonstrates that EEP sensitizes TRAIL-resistant prostate cancer cells. The main phenolic components detected in Brazilian green propolis are artepillin C, quercetin, kaempferol and p-coumaric acid. Brazilian propolis and its bioactive components markedly augmented TRAIL-mediated apoptosis and cytotoxicity in prostate cancer cells. Brazilian EEP enhanced the expression of TRAIL-R2 and the activity of NF-κB in LNCaP cells. The co-treatment of prostate cancer cells with 100 ng/ml TRAIL and 50 µg/ml EEP increased the percentage of apoptotic cells to 65.8 ± 1.2% and caused a significant disruption of ∆Ψm in LNCaP cells. We show that Brazilian EEP helped cells overcome TRAIL resistance by engaging both intrinsic and extrinsic apoptotic pathways and regulating NF-κB activity. The data demonstrate the important role of Brazilian green propolis and its bioactive compounds in prostate cancer chemoprevention through the enhancement of TRAIL-mediated apoptosis.

7-Ketocholesterol and 7-hydroxycholesterol in pork meat and its gravy thermally treated without additives and in the presence of onion and garlic.

The effect of onion and garlic on the formation of two cholesterol oxidation products (COPs): 7-ketocholesterol and 7-hydroxycholesterol was evaluated by comparing their concentrations in meat and gravy samples obtained from three pork dishes prepared in the presence and absence of these flavourings. The concentration of these compounds in meat samples was between 82.4 and 1331.6 ng/g of cooked meat. Gravies contained lower amounts: from 18.3 to 45.6 ng/g of cooked meat. The addition of onion (30 g/100g of meat) caused a decrease in 7-ketocholesterol and 7-hydroxycholesterol concentrations in all of the investigated pork dishes by 9.5-79%, whilst the addition of 15 g of garlic to 100g of meat lowered the concentration by 17 to 88%. The greatest decrease was found in grilled minced chops. The quantitative assessment of 7-ketocholesterol and 7-hydroxycholesterol was carried out by thin-layer chromatography with densitometric detection.

Analysis of azaarenes in pan fried meat and its gravy by liquid chromatography with fluorescence detection.

A method for analysis of six azaarenes (benzo[h]quinoline, benzo[a]acridine, benzo[c]acridine, dibenzo[a,c]acridine, dibenzo[a,j]acridine and dibenzo[a,h]acridine) in thermally treated high-protein food has been described. The clean-up procedure used based on alkaline hydrolysis, tandem solid phase extraction on columns filled with Extrelut - diatomaceous earth and cation exchanger (propyl sulfonic acid), enabled a selective isolation of carcinogenic compounds belonging to benzoacridines and dibenzoacridines from samples of cooked meat and its gravy. The isolated fractions of aza-PAHs were analysed by high-performance liquid chromatography with fluorescence detection. The detection limits for the azaarenes were between 0.0001ng and 0.005ng loaded on column. The recoveries for the four-ring and five-ring azaarenes were from 55% to 67%. Two types of dishes prepared from pork by pan-frying were investigated. Total contents of the benzoacridines and dibenzoacridines determined in cooked meat were 1.57 and 2.50ng/g in collar and chop samples, respectively; their gravies contained 0.34 and 0.59ng of these azaarenes per g of cooked meat.

[Chromatographic methods in tobacco smoke assessment]. / Metody chromatograficzne w ocenie narazenia na dym tytoniowy.

In this paper review of chromatographic techniques and detectors used to analyse nicotine and its metabolites has been presented. The chromatographic determination of xenobiotics in biological material is usually proceeded by multistage isolation procedures, using liquid-liquid and liquid-solid extraction in various modifications. In our investigations, planar chromatography with densitometry has been used to identify and quantify nicotine and its metabolites in body fluids for assessment of tobacco smoke exposure. This technique does not require special cleanup techniques and as well is economic and environmental friendly.

Concentration of chosen oxycholesterols in plasma of pregnant women with pregnancy-induced hypertension.

Solid-phase extraction (SPE) was applied for isolation of oxycholesterols from plasma lipid extract from pregnant women with hypertension and from a control group. Separation of oxycholesterols fraction was performed in an SD II horizontal chamber (Chromdes, Poland) using silica gel and octadecyl RPC18 silica gel TLC plates (Merck and Machery Nagel). Visualization was carried out under UV light after Liebermann-Burchard reaction specific for cholesterol and its derivatives. The oxycholesterols (5-cholestene-3beta-ol-7-one, sum of 5-cholestene-3beta, 7beta-diol and 5-cholestene-3beta, 7alpha-diol and sum of 5alpha,6alpha-epoxycholestan-3beta-ol and 5beta, 6beta-epoxycholestan-3beta-ol) were quantified by chromatograms scanning in reflectance and fluorescence mode using a CS 9301 densitometer (Shimadzu). The total concentration of the investigated oxycholesterols in the plasma of pregnant women was up to 5000 ng/mL and was statistically significantly higher in women with pregnancy induced hypertension (PIH).