Combined assessment of lysine and N-acetyl cadaverine levels assist as a potential biomarker of the smoker periodontitis.

Periodontitis is an inflammatory condition of supporting structures of teeth leading to attachment and bone loss. Cigarette smoking is the single most important and modifiable risk factor with 5 to 20-fold susceptibility for periodontal diseases. Reverse smoking is a peculiar habit of smoking where the lit end is kept inside the mouth, which is predominant in the northern coastal districts of Andhra Pradesh. Polyamines are biologically active amines involved in tissue regeneration and modulation of inflammation. The study aimed to evaluate polyamines and check their utility as a marker in detection of periodontitis among different groups. Total polyamine levels showed significant increase in reverse smokers with periodontitis when compared to the other groups. Qualitative analysis by thin layer chromatography showed three polyamine bands with varying intensity among the different groups. Mass spectrometric and NMR analyses of the three bands identified them as N1, N8-diacetyl spermidine, N-acetyl cadaverine and lysine. Most significantly elevated levels of lysine was observed in the smoker and reverse smoker periodontitis groups when compared to healthy and non-smoker periodontitis groups. The significantly elevated levels of N-acetyl cadaverine could be responsible for the more destruction of periodontium in the reverse smoker group. Antioxidant potential decreased significantly in different smoker periodontitis groups. The present study suggests that the quantitative analysis of salivary polyamines, lysine and N-acetyl cadaverine can aid as an easy noninvasive diagnostic method for assessing the periodontal status, especially in smokers.

Effect of collagen casing on the quality characteristics of fermented sausage.

OBJECTIVE: Fermented sausage is popular all over the world for its rich nutrition and unique flavor. Sausage casing is one of the key factors affecting the quality of fermented sausage. However, there is little information involved in this field. METHODS: In this study, collagen casings were used as a wrapping material, and natural casings (pig casings) were used as a control. The effects of the two types of casings on biogenic amine content and other quality characteristics of fermented sausage were analyzed with increasing the storage time. RESULTS: The results showed that with storage time increasing, the hardness and gumminess of fermented sausage in collagen casing (CC) group were higher than those in pig casing (PC) group (P<0.05), while the elasticity in CC group was lower than that in PC group (P<0.05). In the processing and storage period, there was no significant difference in the type and content of flavor substances between the two groups. More importantly, the contents of tryptamine, putrescine, cadaverine, histamine, tyramine and phenyethylamine in fermented sausage of CC group were lower than those in PC group (P<0.05). CONCLUSION: In conclusion, this study revealed that CC could improve the quality characteristics of fermented sausage and reduce the content of biogenic amines in fermented sausage, which provides a theoretical basis for the choice of casings in industrial production in the future.

The content of biogenic amines in Rondo and Zweigelt wines and correlations between selected wine parameters.

The purpose of this study was to evaluate the content of biogenic amines (BAs) in wines using dispersive liquid-liquid microextraction-gas chromatography-mass spectrometry (DLLME-GC-MS). An additional objective was to assess the correlations between selected parameters characterizing the samples such as the content of BAs, sugars, and organic acids, pH, and total acidity. Wines produced from the same grape variety in which alcoholic fermentation (AF) was carried out by different yeast strains and in which malolactic fermentation (MLF) was spontaneous, differed in the content of biogenic amines. The concentrations of putrescine, cadaverine and tryptamine were higher in the Rondo wines (237-405, 34.04-61.11,

Changes in the microbiological quality and content of biogenic amines in chicken fillets packed using various techniques and stored under different conditions.

The aim of this study was to determine the biogenic amines (BAs) formed in chicken breast meat packaged using different techniques (AP, Hi-O2-MAP or VP) during the storage under different conditions (cold room or display case), to correlate the microbiological quality (TPC, LAB, Pseudomonas spp. and Enterobacteriaceae) of chicken meat with BAs formation and to assess the suitability of selected biogenic amines as indicators of chicken meat spoilage. The initial TPC of chicken fillets was 2.57-3.04 log cfu/g. Over time a systematic significant (p ≤ 0.05) increase in TPC was observed to >7.5 log cfu/g (AP and VP; display case) determined on day 9. It was found that cadaverine and tyramine dominated in quantitative terms in chicken fillets, regardless of packaging technique and storage conditions (166.00 mg/kg in AP meat in cold room on day 9 and 175.03 mg/kg on day 9 in MAP meat in display case, respectively). Taking into account the BAI, high and significant (p ≤ 0.05) correlation coefficients (from 0.51 to 0.95) were obtained with all analyzed indicators of microbiological quality. The concentration of cadaverine, putrescine contents or BAI can potentially serve as chemical quality indicator for freshness of chicken meat.

Natural Contaminants in Wines: Determination of Biogenic Amines by Chromatographic Techniques.

Biogenic amines (BAs) are natural contaminants of wine that originate from decarboxylase microorganisms involved in fermentation processes. The primary relevance of biogenic amines in food could have both toxic effects on consumers' health (i.e., allergic reactions, nausea, tremors, etc.), if present at high concentrations, and concurrently it can be considered as a remarkable indicator of quality and/or freshness. Therefore, the presence of nine biogenic amines [Tryptamine (TRP), ß-phenylethylamine (ß-PEA), putrescine (PUT), cadaverine (CAD), histamine (HIS), serotonin (SER), tyramine (TYR), spermidine (SPD), and spermine (SPM)] was investigated in red and white wine samples, which differed in the winemaking processes. The qualitative-quantitative determination of BAs was carried out by chromatographic methods (HPLC-UV/Vis and LC-ESI-MS). The analysis showed that both winemaking processes had all the nine BAs considered in the study at different amounts. Data showed that red wines had a higher concentration of PUT (10.52 mg L-1), TYR (7.57 mg L-1), and HIS (6.5 mg L-1), the BAs most involved in food poisoning, compared to white wines, probably related to the different type of fermentation (alcoholic and malolactic).

Determination of 8 biogenic amines in aquatic products and their derived products by high-performance liquid chromatography-tandem mass spectrometry without derivatization.

A method for the determination of 8 biogenic amines in aquatic products and their derived products was established by HPLC-MS/MS without derivatization. The samples were extracted by 5% perchloric acid solution. N-hexane was used to clean the extract. The analytes were separated by a column of ACQUITY UPLC HSS T3 (100 mm × 2.1 mm, 1.8 µm), and gradient eluted with a mixed solution of (0.5% formic acid) and acetonitrile. Good linearity was obtained with correlation coefficients (R2) >0.99. This method achieved higher sensitivity (from 0.1 mg/kg for tyramine, 2-phenylethylamine and tryptamine to 1.0 mg/kg for spermidine, spermine, cadaverin, histamine and putrescine). The average recoveries were demonstrated in the range of 70.9%-113.1%, with relative standard deviations (RSDs) from 0.33% to 10.81%. This method was suitable for the detection of BAs in aquatic products and their products.

Three-in-one via syringe needle-based device: sampling, microextraction and peroxidase-like catalysis for colorimetric detection of the change of biogenic amines levels with time in meat.

In this work, a syringe needle-based integrated method was designed for the detection of biogenic amines (BAs) in raw meat samples. Based on a sequential process, the needle-based sampling, micro liquid-phase extraction and peroxidase-like catalysis were adopted for the sample collection, target analytes extraction and colorimetric analysis, respectively. The proposed method exhibited high selectivity towards BAs (the total amount of histamine, putrescine and cadaverine was utilized to present the level of BAs), where the linear range is 5-50 µM and 50-1000 µM, and the limit of detection is 1.52 µM. Specifically, the whole process could be completed in a single syringe needle. In addition, due to the minimized sampling, the change of BAs levels with time in different area of real samples (fish) can be conveniently investigated. This method has the advantages of simplicity, low cost, high sensitivity and selectivity, endowing it a promising candidate for food analysis.

Electrified liquid-liquid interface as an electrochemical tool for the sensing of putrescine and cadaverine.

Putrescine and cadaverine are biogenic amines that serve as potential biomarkers for several types of cancers and monitoring food quality. Electrochemical sensing of putrescine and cadaverine by non-enzymatic routes remains a challenge because of their inertness at unmodified electrode surfaces and hence a liquid-liquid interface strategy has been employed for their detection. In the present study, electrochemical sensing of cadaverine and putrescine has been demonstrated by simple and facilitated ion-transfer processes using a liquid-liquid microinterface supported by a microcapillary. A microinterface was constructed in different configurations by varying the aqueous phase composition in the absence and presence of dibenzo-18-crown-6, and the ion-transfer ability of putrescine and cadaverine was studied in these configurations. A peak shaped voltammogram was observed in the backward scan, due to the linear diffusion of putrescine and cadaverine from the organic to the aqueous phase. The detection ability in the presence of dibenzo-18-crown-6 was observed in the concentration ranges of 0.25-25 µM and 0.25-40 µM for putrescine and cadaverine with detection limits of 0.11 and 0.17 µM respectively. In the presence of dibenzo-18-crown-6, the electrochemical sensing of putrescine and cadaverine was more pronounced compared to the simple ion-transfer process.

Simple and rapid determination of biogenic amines in fish and fish products by liquid chromatography-tandem mass spectrometry using 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate as a derivatization reagent.

A simple and rapid analytical method was developed for determination of four biogenic amines [histamine (Him), cadaverine (Cad), tyramine (Tym), 2-phenylethylamine (Pea)] in fish and fish products. This method uses a new derivatization reagent, 2,4,6-triethyl-3,5-dimethyl pyrylium trifluoromethanesulfonate (Py-Tag). The four biogenic amines in the samples were extracted with trichloroacetic acid. The diluted extract was derivatized with Py-Tag (15 min at 50°C) and then subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The limits of quantification for the method were 2 mg/kg for Him, Tym, and Pea and 10 mg/kg for Cad. The matrix effects derived from the tested fish and fish products were negligible in the LC-MS/MS analysis. The impact of the sample matrices on the Py-Tag derivatization was also negligible. The trueness and repeatability of the method were assessed by performing replicate analyses (n = 5) of five samples of fish and fish products, each spiked with the four biogenic amines at three different concentration levels. Analysis of the samples found 87%-104% of the spiked concentrations and the relative standard deviations were <6.1%. A reference sample and quality control canned fish samples were analyzed by the method, and the concentrations of the Him were within acceptable limits. The developed method was successfully used to determine concentrations of the four biogenic amines in 48 fish and fish products on the Japanese market. The developed method does not require cleanup using a solid-phase extraction column or similar, and the derivatization reaction time was only 15 min. The results suggested that the present method is reliable and suitable for rapid analysis of the four biogenic amines in fish and fish products.

Determination of six underivatized biogenic amines by LC-MS/MS and study of biogenic amine production during trout (Salmo trutta) storage in ice.

Biogenic amines (BAs) are natural components of food produced mainly during metabolism in animals and plants. The determination of BAs is important because of their potential toxicity and their potential use as food spoilage indicators. In the present study, a method for the determination of six BAs (putrescine, cadaverine, histamine, ß-phenylethylamine, tyramine, and tryptamine) by Liquid Chromatography - Tandem Mass Spectrometry (LC-MS/MS) with Atmospheric Pressure Chemical Ionisation (APCI) source has been used on trout samples (Salmo trutta) stored in ice for 15 days. The results showed that on day 15 quite large amounts of putrescine (76.530 mg/kg), cadaverine (85.530 mg/kg), tryptamine (25.210 mg/kg), and histamine (15.975mg/kg) were detected, while the other BAs remained low (ß-phenylethylamine: 3.230 mg/kg, tyramine: 0.165mg/kg). Furthermore, microbiological data (Total Vial Count- TVC, Pseudomonas spp, and Shewanella putrefaciens) showed that trout samples became organoleptically unacceptable on day 12, while volatile compound analysis showed a significant increase in total amounts of alcohols, aldehydes, and ketones on days 12 and 15.

Silver Nanoparticles for a Colorimetric Determination of Putrescine and Cadaverine in Biological Samples.

A convenient and uncomplicated scheme has been projected for the quantitative determination of essential diamines putrescine (PUT) and cadaverine (CAD) via sodium dodecyl sulfate protected silver nanoparticles (SDS-AgNPs). This scheme is based on the chemical interaction of a SDS-AgNPs probe with PUT and CAD, leading to a color change from yellow to red or reddish brown. The interaction was investigated through different techniques such as using a UV-visible spectrophotometer, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), dynamic light scattering spectroscopy (DLS) and the zeta potential. Both amines possess a close resemblance in structure (except for the addition of one more methylene group in CAD), and no any distinguishable color change was noted. However, the maximum absorption band at 580 and 600 nm was demonstrated for PUT and CAD correspondingly. The methodical response was observed at absorption ratios of 580/410 and 600/410 nm, with the linear regression within 4 - 12 and 6 - 14 µg/mL for PUT and CAD. The detection limits calculated for both the diamines PUT and CAD were 0.333 and 1.638 µg/mL. The scheme was successfully applied for determinations in biological samples, including spiked blood plasma and urine. Putrescine exhibited % recovery within 95.717 - 105.200%, while cadaverine was within 95.940 - 105.109%, respectively. The scheme was reproducible and precise with inter-day RSD (n = 5) within 1.126, 0.018% and the intraday RSD (n = 5) was within 0.005, 0.002% for PUT and CAD, respectively.

Exclusive enteral nutrition mediates gut microbial and metabolic changes that are associated with remission in children with Crohn's disease.

A nutritional intervention, exclusive enteral nutrition (EEN) can induce remission in patients with pediatric Crohn's disease (CD). We characterized changes in the fecal microbiota and metabolome to identify the mechanism of EEN. Feces of 43 children were collected prior, during and after EEN. Microbiota and metabolites were analyzed by 16S rRNA gene amplicon sequencing and NMR. Selected metabolites were evaluated in relevant model systems. Microbiota and metabolome of patients with CD and controls were different at all time points. Amino acids, primary bile salts, trimethylamine and cadaverine were elevated in patients with CD. Microbiota and metabolome differed between responders and non-responders prior to EEN. EEN decreased microbiota diversity and reduced amino acids, trimethylamine and cadaverine towards control levels. Patients with CD had reduced microbial metabolism of bile acids that partially normalized during EEN. Trimethylamine and cadaverine inhibited intestinal cell growth. TMA and cadaverine inhibited LPS-stimulated TNF-alpha and IL-6 secretion by primary human monocytes. A diet rich in free amino acids worsened inflammation in the DSS model of intestinal inflammation. Trimethylamine, cadaverine, bile salts and amino acids could play a role in the mechanism by which EEN induces remission. Prior to EEN, microbiota and metabolome are different between responders and non-responders.

Biogenic amine content during the production and ripening of Sremski kulen, Serbian traditional dry fermented sausage.

Sremski kulen is a wide diameter dry fermented sausage, produced from pork, seasoned with red spicy paprika, stuffed into pork cecum, and preserved by smoking, fermentation and drying. Due to specific ripening process, Sremski kulen is suitable for the accumulation of biogenic amines. Therefore, the aminogenesis was studied in traditionally produced Sremski kulen, taking into account the physicochemical parameters and microbial counts. The content of six biogenic amines (tryptamine, phenylethylamine, tyramine, histamine, putrescine, and cadaverine) was analyzed using high-performance liquid chromatography. The ripening process of Sremski kulen was slow followed by changes in aw and pH value as well as expressed proteolysis. The autochthonous microbiota showed pronounced decarboxylase activity. Tryptamine and phenylethylamine were detected at each examined ripening stage while histamine was not detected until the end of ripening (16.55 ± 2.33 mg/kg). Tyramine, cadaverine, and putrescine content significantly increased during the ripening period (p < .05). In the final product, cadaverine was the dominant biogenic amine (132.40 ± 5.05 mg/kg), followed by tyramine (115.80 ± 15.46 mg/kg) and putrescine (68.55 ± 2.39 mg/kg). Although the long ripening period greatly contributed to the accumulation of biogenic amines in final product, their content are not of concern from product safety aspects, but requires improvement in hygiene of production process.

Colorimetric sensor and LDI-MS detection of biogenic amines in food spoilage based on porous PLA and graphene oxide.

Biogenic amines are the important markers for food spoilage, thus, an on-package sensor for biogenic amine detection is crucial for food quality control. A dual detection platform including colorimetry and LDI-MS was developed for screening and quantitative determining of biogenic amines. Porous PLA film, was fabricated using calcium carbonate nanoparticles to enhance film porosity leading to increased surface area of colorimetric sensor. The color intensity significantly increases depending upon the enhanced analyte concentration with a linear range of 2.0-10.0 mg/mL for putrescine, and 0.1-6.0 mg/mL for cadaverine. On another layer, graphene oxide paper was applied as an LDI-MS substrate for sensitive quantification of biogenic amines. LOD values measured on graphene oxide coated side by LDI-MS were found to be 0.07 pM and 0.02 pM for putrescine and cadaverine, respectively. This platform was successfully applied for the detection of biogenic amines in pork samples with satisfactory results.

Process improvement to prevent the formation of biogenic amines during soy sauce brewing.

Biogenic amines (BAs) are a class of bioactive organics produced during the fermentation of soy sauce. A high concentration of BAs may bring about serious physiological and toxicological effects on the human body. In this study, we reported an optimized process to produce soy sauce with lower BA concentration and found the contents of putrescine, cadaverine and histamine increased with the increase of fermentation temperature but decreased with the increase of NaCl concentration. The final content of total BAs with improved fermentation was 105.56 ± 0.13 mg/L, which was reduced by 89.11% compared to traditional brewing. Besides, the pilot production test was performed to verify the optimized conditions and physicochemical indexes were measured to better understand the change principle of the chemical compounds. Taken together, we present an effective process to inhibit the formation of BAs while ensuring that characteristic nutrients are not lost.

Smartphone-interrogated test supports for the enzymatic determination of putrescine and cadaverine in food.

Diamino-oxidase (DAO), horseradish peroxidase (HRP), and tetramethylbenzidine (TMB) have been immobilized into cellulose to obtain circular cellulose test supports (CCTSs) for the determination of cadaverine (Cad) and putrescine (Put). During the enzymatic reaction, TMB is oxidized and a blue spot is obtained. This color (RGB coordinates) is measured with a smartphone and a commercial application. The highest sensitivity is provided by the component R and a linear response is observed for low biogenic amine (BA) concentrations, but a second-order polynomial response better fits the experimental results for a wider concentration range. This has been successfully explained with a model developed to explain the RGB values obtained in this type of analytical system. Optimization studies enable CCTSs to be obtained for Put and Cad determination, which could be used (kept at 4 °C) for at least 45 days if a stabilizer (StabilCoat™ or StabilGuard™) is added during its synthesis. In these conditions, the R coordinate follows the model up to at least 4 × 10-4 M Put and/or Cad (both analytes give the same response). The method permits the Put and Cad determination from 5 × 10-5 M up to 4 × 10-4 M (RSD = 3%, n = 3). The CCTSs have been applied to Put + Cad determination in a tuna sample without any interference by other biogenic amines. The concentration found statistically agrees with that obtained using a HPLC-MS-validated method. Graphical abstract.

Determination of biogenic amines in Chub Mackerel from different storage methods.

The eight biogenic amines (BA), which are histamine (HIS), tryptamine, putrescine, 2-phenylethylamine, cadaverine, tyramine, spermidine, and spermine, were determined in Chub Mackerel under different storage conditions after being freshly caught. The storage time and temperature were varied and the guts either present or removed. This study describes a reverse-phase high-performance liquid chromatography (HPLC) with a fluorescence detector following precolumn derivation with dansyl chloride method for the determination of BA in in Chub Mackerel samples. HIS represented the highest content of BA regardless of the storage temperature and time. During a 24-hr period, the content of HIS reached its highest level of 6,466.63 mg/kg at the storage temperature of 30 °C, whereas it only reached 28.73 mg/kg in 24 hr when the storage temperature was 0 °C, which is way below the standard acceptable threshold level (400 mg/kg). The storage times for the content of HIS to exceed the standard threshold level for HIS at different storage temperatures of 0, 4, 10, 15, 20, and 25 °C were 12, 5, 4 days, 48, 36, 15, and 14 hr, respectively. However, these times were significantly shortened when the viscera was removed from freshly-caught fish before storage. This study not only provides data for monitoring the content of BA, especially HIS, to ensure the safe storage and consumption of freshly-caught Chub Mackerel, but also presents a cost-effective method to extend the storage time of freshly-caught Chub Mackerel. PRACTICAL APPLICATION: This study not only provides data for monitoring the content of BA, especially HIS, to ensure the safe storage and consumption of freshly-caught Chub Mackerel, but also presents a cost-effective method, which is gut-removal during storage, to extend the storage time of freshly-caught Chub Mackerel.

Analytical possibilities of Putrescine and Cadaverine enzymatic colorimetric determination in tuna based on diamine oxidase: A critical study of the use of ABTS.

The joint determination of putrescine (Put) and cadaverine (Cad) in the presence of other biogenic amines is studied using their enzymatic reaction with diamine oxidase (DAO). Three alternative methods are studied based on the intrinsic colorimetric properties of DAO or horseradish peroxidase (HRP), and the use of 2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) colorimetric reagent, respectively. In this last case an in-depth study is carried out in order to explain and solve some drawbacks usually associated with the use of this reagent (especially interferences, interaction with enzymes and instability), and to propose new analytical methodologies which this reagent allows to achieve (transient signal and the use of the violet species). Finally, the method has been applied to Put + Cad determination in a tuna sample without interference of other biogenic amines. The result has been compared with that obtained using a method based on HPLC-MS, which has allowed the new methodology to be validated.

Ultrasensitive, Selective, and Highly Stable Bioelectronic Nose That Detects the Liquid and Gaseous Cadaverine.

Field-effect transistor (FET) devices based on conductive nanomaterials have been used to develop biosensors. However, development of FET-based biosensors that allow efficient stability, especially in the gas phase, for obtaining reliable and reproducible responses remains a challenge. In this study, we developed a nanodisc (ND)-functionalized bioelectronic nose (NBN) based on a nickel (Ni)-decorated carboxylated polypyrrole nanoparticle (cPPyNP)-FET that offers the detection of liquid and gaseous cadaverine (CV). The TAAR13c, specifically binding to CV, which is an indicator of food spoilage, was successfully constructed in NDs. The NBN was fabricated by the oriented assembly of TAAR13c-embedded NDs (T13NDs) onto the transistor with Ni/cPPyNPs. The NBN showed high performance in selectivity and sensitivity for the detection of CV, with excellent stability in both aqueous and gas phases. Moreover, the NBN allowed efficient measurement of corrupted real-food samples. It demonstrates the ND-based device can allow the practical biosensor that provides high stability in the gas phase.

Freshness Evaluation in Chub Mackerel (Scomber japonicus) Using Near-Infrared Spectroscopy Determination of the Cadaverine Content.

We analyzed the volatile basic nitrogen content, pH, total viable cell count, and biogenic amine contents in chub mackerel (Scomber japonicus) stored at 5 and 25°C to examine changes in freshness. Among the various parameters, we found the volatile basic nitrogen content had the highest correlation with cadaverine content (r2 = 0.72 to 0.88). We also tried to measure cadaverine contents at different times during storage by using near-infrared (NIR) spectroscopy. However, because of the high water content in the fish, we could not obtain meaningful results. Next, we prepared samples for NIR spectroscopy by dilution with 0.1 N HCl, ultrafiltration (3 or 10 kDa) with a glass filter, and dehydration. The samples prepared with the 3-kDa filter had peaks in the NIR spectra between 1,379.3 and 1,388.9 nm, and those prepared with the 10-kDa filter had peaks in the spectra between 1,897.3 and 1,898.6 nm. The correlation coefficient (r2) between the NIR spectroscopy and high-performance liquid chromatography with cadaverine results was 0.98 to 0.99. We concluded that the biogenic amine content could be used to evaluate freshness in fish products, and that NIR measurements could be used to rapidly and accurately determine freshness.