Effects and mechanisms of using "clean" smoke particles in the smoking process on the formation of ß-carboline heterocyclic amines (ß-CHAs) in smoked meat patties.

ß-Carboline heterocyclic amines (ß-CHAs), known for their synergistic neurotoxic and carcinogenic effects, are predominantly produced by humans through cigarette smoke and food and are found particularly in meats cooked at high temperatures. Few studies have explored the differences in the mechanisms of accumulation of ß-CHAs in smoked meat and meat processed at high temperatures. In this research, the concentration of ß-CHAs in smoked meats prepared using a variety of wood materials was measured using LCMS/MS. Additionally, key volatile organic compound markers associated with ß-CHAs accumulation in smoke were identified through GCMS and multivariate statistical analysis and subsequently confirmed in a chemical simulation system. Three types of strainers, each with a distinct aperture size, were used to assess the efficacy of particle filtration in reducing ß-CHAs levels in smoked meat. The findings indicated that smoke exposure indeed increases the ß-CHAs content of meat. However, only the strainer capable of filtering PM2.5-sized particles reduced the amount of ß-CHAs present compared to the control group. In contrast, strainers with larger pore sizes facilitated excessive accumulation of ß-CHAs. The presence of aldehydes such as 1 H-pyrrole-2-carboxaldehyde, 5-methylfurfural, benzaldehyde, furfural, and nonanal exhibited a positive correlation with the accumulation of ß-CHAs. Conversely, phenolic compounds, including 2-methoxy-4-vinylphenol, 2-methoxy-5-methylphenol, p-cresol, phenol, 2-methoxy-4-(1-propenyl)-, (Z)-, phenol, 3-ethyl-, and phenol, 4-ethyl-2-methoxy-, showed a negative correlation. Thus, filters made from chelated carbonyl trap materials both chemically and physically disrupt the buildup of ß-CHAs in smoked meats. The use of this approach will not only improve the quality of these products but will also contribute to decreasing the amount of inhalation pollutants released into the environment.

Effect of Chinese quince proanthocyanidins on the inhibition of heterocyclic amines and quality of fried chicken meatballs and tofu.

Heterocyclic amines (HCAs) have potential carcinogenic and mutagenic activity and are generated in cooked protein-rich foods. Adding proanthocyanidins (PAs) to these foods before frying is an effective way to reduce HCAs. In this study, polymeric PAs (PPA) and ultrasound-assisted acid-catalyzed/catechin nucleophilic depolymerized PAs (UAPA, a type of oligomeric PA) were prepared from Chinese quince fruits (CQF). Different levels of PPA and UAPA (0.05%, 0.1%, and 0.15%) were added to chicken meatballs and tofu; then these foods were fried, and the content of HCAs in them after frying was investigated. The results showed that PPA and, particularly, UAPA significantly inhibited the formation of HCAs in fried meatballs and tofu, and this inhibition was dose-dependent. The inhibition of HCAs by both PPA and UAPA was stronger in the chicken meatballs than in fried tofu. The level of total HCAs was significantly reduced by 57.84% (from 11.93 to 5.03 ng/g) after treatment of meatballs with 0.15% UAPA, with inhibition rates of 78.94%, 50.37%, and 17.81% for norharman, harman, and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), respectively. Of note, there was a negative correlation between water, lipid, protein, creatine, and glucose content and HCA content in the crust, interior, and whole (crust-plus-interior) measurements of all fried samples. Interestingly, PPA and UAPA were found more effective in inhibiting HCAs in the exterior crust than in the interior of the fried chicken meatballs. These results provide evidence that further studies on the reduction of the formation of harmful HCAs in fried foods by adding CQF PAs could be valuable to the fried food industry. PRACTICAL APPLICATION: Chinese quince proanthocyanidins treatments significantly inhibited the generation of heterocyclic amines (HCAs) in chicken meatballs and tofu when deep-fried. These results suggest that Chinese quince proanthocyanidins can be used as natural food additive for reducing HCAs in fried foods, laying the foundation for using Chinese quince fruit proanthocyanidins for HCA inhibition in the food industry.

Inhibitory and antioxidative capacity of nutmeg extracts on reduction of lipid oxidation and heterocyclic amines in pan-roasted beef patties.

Identification and inhibition of mutagenic and carcinogenic heterocyclic amines (HCAs) from pan-roasted beef patties were performed by adding (0.02%) tertiary butyl hydroquinone (TBHQ) and (0.05%) ethanol-extracted nutmeg (ENE) using HPLC and principal component analysis. Ten HCAs, including six polar and four non-polar, were assessed. The addition of (0.05%) ENE significantly (P < 0.05) reduced the cooking loss and shrinkage of patties during cooking and reduced the total formation HCAs by 73.97%, which proved the significant (P < 0.05) inhibitory effect as a natural antioxidant against lipid oxidation and HCA formation compared to TBHQ. The DPPH radical-scavenging activity, total phenolic content, and available active metabolites of ENE were estimated. Furthermore, a positive correlation was observed between pH, level of thiobarbituric acid reactive substances, and HCA formation in both the groups. TBHQ and ENE were significant HCAs inhibitors (P < 0.001), but ENE showed resilient oxidative stability during refrigeration storage. Therefore, ENE can be used to reduce HCAs formation in pan-roasted beef patties.

Propolis extract reduces heterocyclic aromatic amine formation in chicken thigh meat.

1. The objective of the present study was to examine the effect of propolis extract on reducing the formation of carcinogenic/mutagenic heterocyclic aromatic amines (HAAs), thereby minimising dietary exposure in human consumers.2. Chicken thigh meat samples were marinated with various concentrations (0%, 0.25%, 0.5% and 1%) of propolis extract, and cooked in a pan at 150°C or 200°C. Proximate composition, pH, lipid oxidation, creatine, creatinine content and twelve HAA levels of samples were analysed.3. Varying levels of IQx (≤35.44 ng/g), MeIQx (≤0.58 ng/g), MeIQ (≤1.60 ng/g), 7,8-DiMeIQx (≤0.83 ng/g), 4,8-DiMeIQx (≤0.75 ng/g), Harman (≤5.54 ng/g), Trp-P-2 (≤1.77 ng/g), PhIP (≤1.61 ng/g) and AαC (≤0.93 ng/g) were quantified in control samples. Total HAA levels ranged between 2.83 and 47.26 ng/g across all samples. Propolis extract decreased the levels of total HAAs by 41.2-89.4% and 49.4-91.4% at 150°C and 200°C, respectively.4. The results demonstrated that propolis extract marination might be an effective strategy to reduce the dietary exposure of HAAs via mitigating their formation in chicken thigh meat.

Effect of Different Roasting Conditions and Coreopsis Extract on Heterocyclic Amine Formation in Roast Lamb Products.

ABSTRACT: Heterocyclic amines (HCAs), which are known carcinogens in thermally processed foods, were investigated in roast lamb patties under various time and temperature conditions. HCAs in lamb products roasted at some temperatures increased with roasting time. An exponential model with a time factor fit well for the production of HCAs. The mean pH and cooking loss at various temperatures were also determined. The mean pH decreased as the temperature increased. Coreopsis extract was added to lamb patties roasted at 230°C for 15 min per side. The amount of coreopsis extract added had a significant effect on HCA development. A weak positive relationship was observed between the antioxidant activity of the lamb patty with the coreopsis extract and the inhibitory effect of coreopsis extract on various HCAs, with a correlation coefficient of 0.14 to 0.44 (P > 0.05). Coreopsis extract containing flavonoids can be a beneficial additive for production of barbecue meat.

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.

Assessment of dietary exposure to heterocyclic amines based on the Korean total diet study.

Heterocyclic amines (HCAs) are contaminants in proteinaceous foods produced by cooking at high temperatures. This study was the first assessment of exposure to HCAs using the Korean total diet study. Twelve HCAs were analysed in 1,232 pooled samples using six isotope-labelled internal standards and HPLC-MS/MS. The daily intake of HCAs in the Korean population was estimated based on the concentration of HCAs in the total diet study samples and individual food consumption data from the Korean National Health and Nutrition Examination Survey. Among HCAs, the intake of ß-carbolines, such as harman and norharman, was the highest, followed by the intake of PhIP. The primary sources of HCA intake were meat, fish, shellfish, and beverages, including alcohol. The margin of exposure to PhIP was 2,349,000 at the average level and 373,000 at the 95th percentile in the Korean population. The estimated daily intake of all HCAs in the Korean population was considered safe.

Chlorogenic acid and Epicatechin: An efficient inhibitor of heterocyclic amines in charcoal roasted lamb meats.

There are few studies on controlling the formation of heterocyclic amines (HAs) by adding polyphenols to roasted lamb. The aim of this study was to assess the inhibitory effect of 0.025, 0.125, and 0.625 mmol of chlorogenic acid, epicatechin, rutin, quercetin and quinic acid on the formation of HAs in charcoal roasted lamb, respectively, by UHPLC-MS/MS. The results indicated that Harman, Norharman, PhIP, IQx and 8-MeIQx were detected, but both chlorogenic acid and epicatechin greatly inhibited the formation of IQx, 8-MeIQx, Norharman, Harman and PhIP, and epicatechin had better inhibitory effect than chlorogenic acid. PLSR-VID analysis suggested the key precursors of HAs formation in roasted lamb were glucose, ribose, fructose, isoleucine, valine, and lysine. In addition, this work also implied that the potential mechanism of inhibition of HAs formation by chlorogenic acid and epicatechin in roasted lamb might attribute to competitive chemical reactions between polyphenols and key precursors of HAs.

Mitigation of heterocyclic amines by phenolic compounds in allspice and perilla frutescens seed extract: The correlation between antioxidant capacities and mitigating activities.

The effect of different levels of allspice and perilla frutescens seed extract (ASE and PSE) on the formation of heterocyclic amines (HCAs) in pan-fried chicken meat patties and the bioactive components found in ASE and PSE that contribute to the mitigation of HCAs were investigated in this study. DPPH radical scavenging activity was evaluated and the results indicated that APSE (ASE + PSE) showed the highest capacity to scavenge free radicals, and the most effective inhibition of HCAs formation. Furthermore, Single and mixed phenolic compounds exhibited a positive effect in scavenging free radicals and mitigating HCAs. The radical scavenging activity and HCAs inhibition effect of single phenolic compounds were highly correlated, whereas mixed phenolic compounds exhibited poor correlation. PCA analysis indicated that phenolic compounds had the maximum inhibitory effect on IQ, followed by Norharman and harman and the minimal effect on PhIP and 7,8-DiMeIQx.

Formation and Prediction of PhIP, Harman, and Norharman in Chemical Model Systems Containing Epicatechin under Various Reaction Conditions.

The aim of this study was to investigate the formation rules of 1-methyl-6-phenylimidazo[4,5-b]-pyridine (PhIP), 1-methyl-9H-pyrido[3,4-b]-indole (Harman), and 9H-pyrido[3,4-b]-indole (Norharman) in chemical model systems containing epicatechin under various reaction conditions and establish their prediction models. The results indicated that at 100-200 °C and pH 5.5-7.5 for 10-90 min, epicatechin inhibited the formation of PhIP, Harman, and Norharman by 32-100, 1-93, and 5-98%, respectively. The exponential growth equation in the growth model with R2 of 0.986-0.997, the second-order equation in the polynomial model with R2 of 0.900-0.949, and the trigonometric function equation in the Fourier model with R2 of 0.513-0.926 were well fitted for the formation of PhIP, Harman, and Norharman at various temperatures, times, and pHs, respectively. The calibration set (R2c) and the prediction set (R2p) of the optimal partial least squares regression prediction model was 0.74 and 0.70 for PhIP, 0.75 and 0.70 for Harman, and 0.90 and 0.91 for Norharman, respectively. The results provide theoretical support to develop technologies or equipment for the real-time synchronous monitoring of PhIP, Harman, and Norharman during meat processing.

Food security, obesity, and meat-derived carcinogen exposure in US adults.

Risk for colorectal cancer (CRC) is increased in adults with poor diet quality, low socioeconomic status, and increased body mass index (BMI). Cooked meats contain high contents of mutagenic compounds related to CRC risk. To explore differences in meat-based carcinogen exposure, a 99-item Qualtrics survey was issued to 1648 US adults. Average monthly serving size, degree of doneness, and cooking methods of meat products were obtained. The National Cancer Institute CHARRED database was used to quantify exposure to HCAs, PAHs, and Ames Predicted and Estimated mutagenicity. Questions from validated instruments assessed food security status (FSS) and demographic variables, while height and weight were self-reported to calculate BMI. Sex, FSS and obesity status (BMI > 30 kg/m2) were compared using two-sample t-tests and multivariate regression models to determine differences in meat intake and carcinogen exposure. Statistical significance was set at P < 0.05. Eight hundred fifty-six valid responses (57.4% female and 81.8% white) were obtained. Non-obese males consumed more white meat and were exposed to greater amounts of 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline. Food insecure and obese females consumed more red meat and had higher exposure rates of several carcinogens. Pan-frying and BBQ were the primary sources of meat-derived carcinogens. Greater disparities in carcinogen exposure were observed in females regarding BMI and FSS. Public health messages to modify meat cooking methods may be most effective for CRC prevention.

Heterocyclic Amine Formation and Mitigation in Processed Meat and Meat Products: A Mini-Review.

ABSTRACT: This review provides an assessment of heterocyclic amine (HCA) formation and mitigation in processed meat and meat products. HCAs are formed when amino acids react with creatine during thermal processing of meat and meat products. The formation of HCAs depends on various factors, including the temperature, cooking time, fat contents, and presence of HCA precursors such as water, lipids, and marinades. Additional factors that could affect HCA formation are pH, meat type, and ingredients added during cooking such as antioxidants, amino acids, ions, fat, and sugars, which promote production of HCAs. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline, 2-amino-3-methylimidazo-[4,5-f]quinoline, and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline are HCAs of importance because of their link to cancer in humans. More than 25 HCAs have been identified in processed foods. Of these, nine HCAs are possible human carcinogens (group 2B) and one is a probable human carcinogen (group 2A). To mitigate HCA generation during heat processing, various techniques have been used, including recipe variations, adjustments of thermal processing conditions, addition of flavorings, pretreatments such as microwave heating, and addition of naturally occurring and artificial antioxidants.

Evaluating the effects of temperature and time on heterocyclic aromatic amine profiles in roasted pork using combined UHPLC-MS/MS and multivariate analysis.

In this study, ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) combined with principal component analysis (PCA) were used to investigate the effects of process conditions on the profiles of carcinogenic and mutagenic heterocyclic aromatic amine (HAA) in the pork roasted at 175 °C, 200 °C, 225 °C and 250 °C for 10, 15, 20, 25, 30, 35 and 40 min. Twelve HAAs from four categories, including carboline (Norharman, Harman, and Phe-p-1), imidazopyridine (PhIP, 4'-OH-PhIP, DMIP, and 1,5,6-TMIP), imidazoquinoline (IQ, IQ [4,5-b], and MeIQ), and imidazoquinoxaline (MeIQx and 4,8-DiMeIQx), were detected, quantified and used to compose the HAA profiles in roasted pork. After being Analyzed by PCA, the distributions of HAA profiles from different temperature on the PCA score plot demonstrated that there are significant differences among the HAA profiles from different temperatures. The loading plot also showed that PhIP, 4'-OH-PhIP, IQ[4,5-b], and MeIQ were mainly responsible for the difference. The profiles from higher temperature distribute more scattered than the lower ones, illustrating that the time effects on the HAA profiles from higher temperature are stronger than the lower ones. Comparing the score and loading plots of different heating times, the diversities of the HAA profiles at different temperatures increased under prolonged heating because of the changingpyridines levels. The results of PCA that comparing the HAA from different categories displayed that the formation features of four categories HAAs were significantly differed because of their formation discrepancy under low temperatures and short-term roasting. Using HAA profiles as an entirety, these findings obtained in this study are more close to the real process of HAA formation in roasted pork, and make the complex effects of temperature and time on multiple HAA formations more simply to be concluded.

An accurate, rapid, and sensitive method for simultaneous determination of four typical heterocyclic amines in roasted pork patties: Application in the study of inhibitory effects of astaxanthin.

Heterocyclic aromatic amines, as a group of mutagenic and carcinogenic compounds, have gained worldwide concern. In this study, an accurate, rapid, and sensitive confirmation and quantification method of four major heterocyclic aromatic amines in roasted pork was developed based on Q-Orbitrap along with Quick, Easy, Cheap, Effective, Rugged, and Safe extraction. The limit of detections and limit of quantitations were found to be 0.2-1.2 µg/kg and 0.6-3.5 µg/kg, respectively, revealing the high sensitivity of this method. Obtained results showed recoveries ranging from 78.1 to 97.4%, depending on the different heterocyclic aromatic amines and spiked levels. Precision was in the range of 2.6-4.5% for four heterocyclic aromatic amines at different levels. In addition, the developed method had been applied to investigate the inhibitory effects of astaxanthin on the above-mentioned heterocyclic aromatic amines in roasted pork. The amount of astaxanthin with the best inhibitory effects was 7.5 mg (0.0375%), which led to significant reduction in heterocyclic aromatic amines levels over 50%.

Effects of seasoning on the formation of heterocyclic amines and polycyclic aromatic hydrocarbons in meats: A meta-analysis.

Cooking improves digestibility, palatability, and microbiological profile of meats, but can produce compounds with carcinogenic potential, such as heterocyclic amines (HAs) and polycyclic aromatic hydrocarbons (PAHs). It has been shown that the formation of these compounds in meats can be inhibited by spiced marinades, but there is a complexity to check and compare the results of isolated studies with so many variables involved. Thus, this work aimed to review studies that evaluated the effects of spices on the formation of HAs and PAHs in meats according to cooking techniques and spice type. A meta-analysis with a random effect model was conducted using response ratios (R) to identify and summarize previous results and answer the research question. The use of spices before frying (R* = 0.52), grilling (R* = 0.63), or roasting (R* = 0.74) meat, and spicing with garlic and onion (R* = 0.57), pepper (R* = 0.63), and other spices with phenolic compounds (R* = 0.63), decrease the formation of HAs and PAHs, due to the antioxidant and electron transfer capacity. In this article, we discuss how the improvement of culinary techniques with the dissemination of knowledge about meat preparation conditions is an effective strategy for reducing the formation and ingestion of HAs and PAHs; this is important due to the growing evidence about the association between meat consumption and chronic diseases. This is the first systematic review with meta-analysis about this topic and can guide industry, food services, and population to improve the safety associated with meat consumption.

The Inhibitory Effects of Heterotrigona Itama Honey Marinades on the Formation of Carcinogenic Heterocyclic Amines in Grilled Beef Satay.

Heterocyclic amines (HCAs) are carcinogenic food toxicants formed in cooked meats, which may increase the risk of cancer development in humans. Therefore, in this study, the effect of stingless bee honey from different botanical origins on the formation of HCAs in grilled beef satay was investigated. HCAs concentration in grilled beef satay was determined by using high performance liquid chromatography (HPLC). In total, six of the most toxigenic HCAs representing aminoimidazo-azaarenes (AIAs) (MeIQx, 4,8-DiMeIQx, and PhIP) and amino carbolines (norharman, harman, and AαC) groups were identified in all the beef samples investigated. A significant reduction in HCAs was observed in grilled beef marinated in honey as compared to beef samples marinated in table sugar (control), in which the reduction of 95.14%, 88.45%, 85.65%, and 57.22% was observed in gelam, starfruit, acacia, and Apis honey marinades, respectively. According to the partial least squares regression (PLS) model, the inhibition of HCAs in grilled beef was shown to be significantly correlated to the antioxidant activity (IC50) of the honey samples. Therefore, the results of this study revealed that the addition of stingless bee honey could play an important role in reducing HCAs in grilled beef.

Structure characteristics of flavonoids for heterocyclic aromatic amines inhibition using quantitative structure-activity relationship modeling.

The objective of this study was to investigate the structure characteristics of flavonoids that act as inhibitors for heterocyclic aromatic amines (HAAs) formation. Five quantitative structure-activity relationship models for predicting the inhibitory rates of HAAs (norharman, harman, PhIP, MeIQx, and 4,8-DiMeIQx) were established using selected chemometric parameters (R2 : 0.591-0.920), and indicated that the hydrophobicity, hydroxyl groups, and topological structure of flavonoids played important roles in the inhibition of HAAs formation. The 5,7-dihydroxyls in meta-position of the A-ring and the 4'-hydroxyl in the B-ring of flavonoids were critical for the inhibitory effects of HAAs, whereas the introduction of 3-hydroxyl and 3-O-glucoside in the C-ring reduced the inhibitory effects. Catechin served as the most effective inhibitor of HAAs followed by luteolin and genistein. The study can bring us a broader idea for controlling the formation of HAAs according to the structure of flavonoids. PRACTICAL APPLICATIONS: Heterocyclic aromatic amines (HAAs) are a class of organic substances with carcinogenic and mutagenic effect formed during the heating process of meat products. The formation of HAAs can be inhibited by adding natural antioxidants such as flavonoids to the meat during pretreatment. This inhibition is influenced by the unique structure of flavonoids. Thus, there has been an increasing demand to exploit the effective HAAs inhibitors from flavonoids by structure characteristics. Our study showed that the inhibitory effect of flavonoids on the formation of HAAs was mainly depended on their hydrophobicity, hydroxyl groups, and topological structure using the multiple QSAR models. Thus, effective HAAs inhibitors can be explored from dietary flavonoids according their structure characteristics.

Heterocyclic Amine Formation in Grilled Chicken Depending on Body Parts and Treatment Conditions.

Heterocyclic amines (HCAs) carcinogenicity is known since the 1970's, but the exact way of their formation is still unclear. During these examinations different body parts (breast filet with and without skin, thigh filet without skin and full wing with skin) of chickens from the same Ross 308 strain were analyzed after grilling with the combination of 3-3 temperature and duration levels (150-180-210 °C and 2.5-5-10 min per side). Five different kinds of heterocyclic amines (HAR, NOR, MeIQx, 4,8-DiMeIQx and PhIP) were detected by HLPC-MS/MS. The results obtained from the present study confirm that, in general, the higher the temperature and longer the duration of the grilling the more HCAs will be generated. Grilling of chicken thigh without bones and skin resulted in lower amounts of HCAs generated in comparison to the grilling of chicken breast without skin. The presence of skin on the chicken breast increased the amounts of HCAs formed, especially if grilling was performed at high temperature for longer duration, especially at 210 °C for 10 min. In case of grilling the chicken wings, the amounts of HCAs formed were lower than observed in the breast.

Antimutagenic effect of an Asclepias subulata extract against heterocyclic aromatic amines commonly found in cooked meat and its heat stability.

The antimutagenicity of an extract from the medicinal plant Asclepias subulata (ASE) against heterocyclic aromatic amines (HAAs) commonly found in cooked meat, as well as its stability to heat treatment (HT), was evaluated. HT (180 °C/3 min) had no effect on the content in ASE of the bioactive compound corotoxigenin-3-O-glucopyranoside; conversely, calotropin significantly decreased by 72%. ASE exerted antimutagenicity against PhIP, MelQ, and MelQx in TA98 and TA100 Salmonella strains, and this activity was not affected by heat, with the exception of MelQ (p < 0.05). Since HAAs can induce colorectal cancer, the thermal stability of ASE's antiproliferative effect against colorectal cancer cells was also evaluated. HT decreased (p < 0.05) the antiproliferative activity of ASE; however, the remaining activity was still strong with an IC50 of 16.8 ± 2.03 µg/mL. Therefore, ASE can be used as a food ingredient to reduce the carcinogenic potential of thermally induced HAAs.

A Systematic Study of Coumarin-Tetrazine Light-Up Probes for Bioorthogonal Fluorescence Imaging.

Fluorescent probes that light-up upon reaction with complementary bioorthogonal reagents are superior tools for no-wash fluorogenic bioimaging applications. In this work, a thorough study is presented on a set of seventeen structurally diverse coumarin-tetrazine probes that produce fluorescent dyes with exceptional turn-on ratios when reacted with trans-cyclooctene (TCO) and bicyclononyne (BCN) dienophiles. In general, formation of the fully aromatic pyridazine-containing dyes resulting from the reaction with BCN was found superior in terms of fluorogenicity. However, evaluation of the probes in cellular imaging experiments revealed that other factors, such as reaction kinetics and good cell permeability, prevail over the fluorescence turn-on properties. The best compound identified in this study showed excellent performance in live cell-labeling experiments and enabled no-wash fluorogenic imaging on a timescale of seconds.