Evaluation of total aflatoxin,nitrate and nitrite levels in layer feed samples of companies producing their own feed in Edincik and Bandırma province of Turkey

Feed contamination by fungi can lead to nutrient losses and detrimental effects on animal health and production. The presence of nitrates and nitrites in food can be harmful to both people and animals. The aim of this study was to determine total aflatoxin, nitrate and nitrite levels in layer feed samples from companies producing their own feed in Edincik and Band?rma provinces in Turkey and to discuss the potential risk to animal health. The results of the analyses indicated that mean total aflatoxin (AFT) ranged from 0.4 to 36.8?µg?kg^?1 and from 0.45 to 47.0?µg?kg^?1 in the year 2007 and the year 2008 samples, respectively. It was determined that nitrate levels were 2.4–10 and 1.7–13?µg?kg^?1 and that nitrite levels were 0–2.4?µg?kg^?1 and 0–2.6?µg?kg^?1 in these years, respectively. The levels of total aflatoxin, nitrate and nitrite in the layer samples could not be considered a risk to poultry health and productivity.     

Dimethylnitrosamine levels in untreated herring meals

An investigation has been conducted to determine the levels of dimethylnitrosamine in 13 samples of Canadian herring meals taken from the Atlantic and Pacific coasts. The concentration of dimethylnitrosamine found in these meals, which had not been treated with nitrate or nitrite varied between 0.15 and 1.0 parts/10⁶.     

Estimated dietary intake of nitrite and nitrate in Swedish children

This study examines the intake of nitrate and nitrite in Swedish children. Daily intake estimates were based on a nationwide food consumption survey (4-day food diary) and nitrite/nitrate content in various foodstuffs. The mean intake of nitrite from cured meat among 2259 children studied was 0.013, 0.010 and 0.007?mg?kg^?1?body?weight?day^?1 in age groups 4, 8–9 and 11–12 years, respectively. Among these age groups, three individuals (0.1% of the studied children) exceeded the acceptable daily intake (ADI) of 0.07?mg?nitrite?kg^?1 body weight?day^?1. The mean intake of nitrate from vegetables, fruit, cured meat and water was 0.84, 0.68 and 0.45?mg?kg^?1 body weight?day^?1 for children aged 4, 8–9 and 11–12 years, respectively. No individual exceeded the ADI of 3.7?mg?nitrate?kg^?1 body weight?day^?1. However, when the total nitrite intake was estimated, including an estimated 5% endogenous conversion of nitrate to nitrite, approximately 12% of the 4-year-old children exceeded the nitrite ADI. Thus, the intake of nitrite in Swedish children may be a concern for young age groups when endogenous nitrite conversion is included in the intake estimates.     

Nitrate and nitrite levels in commonly consumed vegetables in Hong Kong

Levels of nitrate and nitrite in 73 different vegetables, a total of 708 individual samples grouped into leafy, legumes, root and tuber, and fruiting vegetables, which are traded mainly in Hong Kong, were measured. Where available, five samples of each vegetable type were purchased from different commercial outlets during the winter of 2008 and summer of 2009. Levels of nitrate and nitrite were determined by ion chromatography and flow injection analysis, respectively. Nitrate and nitrite levels of all samples ranged <4–6300 and <0.8–9.0 mg?kg^?1, respectively. Nitrate concentrations for the different groups, in descending order, were leafy?>?root and tuber?>?fruiting and legume vegetables. More than 80% of vegetables had mean nitrate concentrations less than 2000?mg?kg^?1, but mean nitrate concentrations of three types of leafy vegetables, namely Chinese spinach, Shanghai cabbage and Chinese white cabbage, were >3500?mg?kg^?1. On the other hand, nitrite concentrations were generally low –?<1?mg?kg^?1 on average. Nitrate in vegetables (i.e. Chinese flowering cabbage, Chinese spinach and celery) can be reduced significantly (12–31%) after blanching for 1–3?min, but not after soaking.     

Fate of Ingested 15N-Labelled Nitrate and Nitrite in the Rat

The fate of ingested nitrate and nitrite was investigated in rats by carrying out single and multiple dose feeding studies with ¹⁵N-labelled nitrate and nitrite. By 72 hr after discontinuation of administration of the ¹⁵N-labelled compounds, about 60–70% of the dose was excreted in the urine, and 10–20% was eliminated in the feces. Residual ¹⁵N remaining in the bbody carcass was about 10% following single dose experiments and 3–4% following continuous feeding trials. One-half or more of the excreted ¹⁵N was not analyzable chemically as nitrate or nitrite. On the basis of chemical analysis, about 30% of the dose was excreted in urine as nitrate plus nitrite; excretion in the feces was low. Negligible amounts of nitrate-N or nitrite-N were found in blood and organs of the animals. Most of the nonnitrate-¹⁵N and nonnitrite-¹⁵N was excreted, like nitrate-N and nitrite-N, rather rapidly from the body.     

Photochemical attenuation of N-nitrosodimethylamine (NDMA) and other nitrosamines in surface water

The aqueous photolysis of seven alkyl nitrosamines was studied by irradiation in a solar simulator. Nitrosamines included N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosodi-n-propylamine (NDPA), N-nitrosodi-n-butylamine (NDBA), N-nitrosopiperidine (NPip), and N-nitrosopyrrolidine (NPyr). Direct photolysis at irradiations of 765 W/m2, representing Southern California midsummer, midday sun, resulted in half-lives of 16 min for NDMA and 12-15 min for the other nitrosamines. The quantum yield for NDMA was determined to be phi = 0.41 and phi = 0.43-0.61 for the other nitrosamines. Quantified products of NDMA photolysis included methylamine, dimethylamine, nitrite, nitrate, and formate, with nitrogen and carbon balances exceeding 98 and 79%, respectively. Indirect photolysis of nitrosamines in surface water was not observed; increasing dissolved organic carbon (DOC) slowed the NDMA photolysis rate because of light screening. Removal of NDMA measured in tertiary treated effluent flowing in a shallow, sunlit engineered channel agreed with photolysis rates predicted based on the measured quantum yield and system parameters. Because biodegradation is relatively slow, aquatic photolysis of NDMA is generally expected to be more significant even at relatively low levels of solar irradiation (t(1/2) = 8-38 h at 244-855 W/m2, 51 degrees N latitude, 1 m depth).     

EFFECTS OF INITIAL NITRITE LEVEL, HEATING AND STORAGE ON RESIDUAL NITRITE AND SPOILAGE OF CANNED HAM ROLL

Canned ham rolls with initial nitrite levels of 0, 125, 200, 300 and 400 mg/kg were processed to an Fo of 1.07 and 1.49 and stored for 8 months. Ham rolls before processing contained 10⁵–10⁷ cfu/g total bacteria and less than 3 MPN/g of putrefactive anaerobic bacterial spores. The initial nitrite level, the storage time and the heat treatment significantly affected residual nitrite in the product. Samples stored for 1 and 3 days had significantly higher residual nitrite levels than those stored for 6–31 days. Samples processed with 0, 125 and 200 mg/kg nitrites contained significantly lowered residual nitrite levels than those formulated with 300 and 400 mg/kg nitrite. Samples heated to an Fo of 1.07 contained less residual nitrite (P ≤ 0.05) than those heated to Fo 1.49. No swelled cans were observed after 8 months of storage and the nitrosamine levels detected in the samples were lower than the safety limit proposed by Pneussman (1974).     

Nitrate and nitrite content in organically cultivated vegetables

The nitrate and nitrite content of leaf vegetables (Swiss chard, sea beet, spinach and cabbage), “inflorescence” vegetables (cauliflower) and fruit vegetables (eggplant and vegetable marrow) grown with organic fertilizers have been determined by a modified cadmium–Griess method. Samples were purchased from organic food stores as well as collected directly from an organic farm in Madrid (Spain). Nitrate levels were much higher in the leaf vegetables (especially Swiss chard species; average over the different samples and species of 2778.6 ± 1474.7 mg kg^{? 1}) than in inflorescence or fruit products (mean values between 50.2 ± 52.6 and 183.9 ± 233.6 mg kg^{? 1}). Following Swiss chard species, spinach (1349.8 ± 1045.5 mg kg^{? 1}) showed the highest nitrate content, and nitrite was found above the limit of detection in some samples only (spinach, 4.6 ± 1.0 mg kg^{? 1}; sea beet, 4.2 ± 0.7 mg kg^{? 1} and Swiss chard, 1.2 ± 0.4 mg kg^{? 1}). Some vegetables (spinach, cabbage and eggplant) had lower nitrate content in the samples harvested in summer, showing the influence of climatic conditions on the nitrate levels in a plant. The samples taken directly from the organic farm, with the exception of eggplant, had higher or slightly higher average nitrate values than samples purchased in the organic food stores, ranging from 117 to 1077%.     

Nitrate Analysis in Meats; Comparison of Two Methods

A high performance liquid chromatographic method (HPLC) was compared to cadmium reduction-Griess (Cd-Griess). Nitrate and nitrite were measured in fresh and cured meats. Nitrate levels by HPLC ranged from 6.2 to 26.7 nmol/g in fresh meats and from 124 to 3018 nmol/g in cured meats. Nitrate contents by HPLC were significantly higher (p<0.05) than those by Cd-Griess. Small amounts of nitrite (0–7.3 nmol/g) were detected in fresh meat samples by Cd-Griess. Results were similar to those used by the National Academy of Sciences to estimate human exposure to nitrate from fresh meats. Results from HPLC methods may provide more accurate estimates of human exposure to nitrate and nitrite.     

Nitrite and nitrate content in meat products and estimated intake in Denmark from 1998 to 2006

The content of nitrite and nitrate in cured meat products has been monitored in Denmark seven times between 1995 and 2006. The maximum permitted added amounts of sodium nitrite in Denmark (60 mg kg^?1 for most products up to 150 mg kg^?1 for special products) have not been exceeded, except for a few samples back in 2002. The intake, mean and intake distribution of sodium nitrite have been calculated from 1998 to 2006 with data from the Danish dietary survey conducted in 2000–02 on Danes from four to 75 years of age. The amounts used by industry have been relatively stable through the whole period with levels varying between 6 and 20 mg sodium nitrite kg^?1 with sausages, meat for open sandwiches and salami-type sausages being the greatest contributors. The mean intake of sodium nitrate was around 1 mg day^?1, which is very low compared with the total intake of 61 mg day^?1. The mean intake of sodium nitrite was 0.017 and 0.014, 0.009 and 0.008, and 0.007 and 0.003 mg kg^?1 body weight day^?1 for men and women in the age groups 4–5, 6–14 and 15–75 years, respectively, which was much lower than the acceptable daily intake (ADI) of 0.09 mg kg^?1 body weight day^?1. The 99th percentile for the group of 4-year-olds was 0.107 and 0.123 mg kg^?1 body weight day^?1 for boys and girls, respectively, and the 95th percentile was 0.057 and 0.073 mg kg^?1 body weight day^?1 for boys and girls, respectively, highest for the girls. With fewer than 100 boys and girls in the 4–5-year age group, only very few persons were responsible for the high intake. The conversion of nitrate to nitrite in the saliva and the degradation of nitrite during production and storage must also be considered when evaluating the intake of nitrite.     

Naturally occuring nitrate/nitrite in foods

A review of the occurrence of nitrate in unprocessed foods showed that high concentrations are frequently found in vegetables. Spinach and lettuce commonly exceed 1000 parts/10⁶ nitrate while, of the root vegetables, beets, turnips and radishes are similarly rich in nitrate. In general, the pulses contain less nitrate than leaf and root vegetables, and most fruits contain little nitrate. Fresh dairy produce is usually low in nitrate. Well waters in some agricultural areas may contain nitrate concentrations so high as to represent a hazard if used as the sole water source for infants, but municipal supplies in the UK rarely exceed 45 parts/10⁶ although they frequently approach this. It is concluded that vegetables and water supplies make a greater contribution to the mean weekly nitrate intake than do cured meats. Nitrite concentrations are usually very low in fresh, undamaged fruits, vegetables and drinking water but adverse storage conditions can lead to (mainly microbial) reduction of nitrate. In these circumstances vegetables naturally rich in nitrate may accumulate nitrite at concentrations which can be toxic to infants.     

Effects of N-nitrosopyrrolidine, nitrite and pyrrolidine on tumour development in mice as related to ingestion of cured meat

The incidence of tumours in mice fed a standard chow diet and given either N-nitrosopyrrolidine (NPyr), nitrite (NO₂) or nitrite plus pyrrolidine (NO₂ + Pyr) in the drinking water for 12 months at 100mg, 1 g and 1 g plus 100mg/litre, respectively, was compared with that for a control group (C) receiving no additives. Differences between groups in weight gain and feed consumption were not significant. Group 3 (NO₂) drank less water (P < 0·05) than those in groups 1 (C) and 2 (NPyr). Water intake of mice in group 4 (NO₂ + Pyr) did not differ from that of any of the other three groups. Survival rates were: 94% (C), 80% (NPyr), 92% (NO₂) and 83% (NO₂ + Pyr); but the differences were not statistically significant. Gross examination upon autopsy revealed that the incidence of all tumours in group 2 (NPyr) was 10- to 20-fold higher than that in any other group. Histological examination confirmed that NPyr induced more (P < 0.05) malignant tumours in liver and lungs with any other treatment; otherwise there were no significant differences. Results indicated that NO₂ + Pyr (nitrite plus a secondary amine) did not increase the frequency of carcinogenic tumours in mice.     

A Micro-Flow-Batch Analyzer Using an In-line Cadmium Sponge Microcolumn for the Photometric Determination of Nitrate and Nitrite in Dairy Samples

In this study, a micro-flow-batch analyzer (μFBA) using an in-line cadmium reduction microcolumn for the photometric determination of nitrate and nitrite in dairy samples is described. The method is based on the Griess-Llosvay reaction and measuring of the absorbance at 540 nm using a green LED integrated into the μFBA built in the urethane-acrylate resin. Initially, the nitrite content of the dairy sample is analyzed in the mixing chamber, while the nitrate is reduced to nitrite in cadmium sponge microcolumn coupled to the microsystem. Nitrate content was calculated from the difference between nitrate plus nitrite (expressed as nitrite) and nitrite content. The analytical curve for nitrate and nitrite was linear in the work range of 10.0–100.0 μg L^?1 with a correlation coefficient of 0.992 and 0.998, respectively. The limit of detection and relative standard deviation were estimated at 0.39 μg L^?1 and <1.7 % (n?=?5) for nitrite and 0.41 μg L^?1 and <1.3 % (n?=?5) for nitrate. Comparing with the reference methods, no statistically significant differences were observed when applying the paired t test at a 95 % confidence level. The accuracy was assessed through recovery test (97.7 to 102.9 %). The proposed microsystem-employed in-line cadmium sponge microcolumn presented satisfactory portability, robustness, flexibility, low-cost device, and reduced chemicals consumption compared to recent methods. Thus, μFBA is potentially useful as an alternative for other automatic determinations using in-line pretreatment steps.     

Prevalence of Nitrite and Nitrate Contents and Its Effect on Edible Bird Nest's Color

Edible bird nests (EBNs) are important ethnomedicinal commodity in the Chinese community. Recently, But and others showed that the white EBNs could turn red by vapors from sodium nitrite (NaNO₂) in acidic condition or from bird soil, but this color‐changing agent remained elusive. The aim of this study was to determine the prevalence of nitrite and nitrate contents and its affects on EBN's color. EBNs were collected from swiftlet houses or caves in Southeast Asia. White EBNs were exposed to vapor from NaNO₂ in 2% HCl, or bird soil. The levels of nitrite (NO₂^?) and nitrate (NO₃^?) in EBNs were determined through ion chromatography analysis. Vapors from NaNO₂ in 2% HCl or bird soil stained white bird nests to brown/red colors, which correlated with increase nitrite and nitrate levels. Moreover, naturally formed cave‐EBNs (darker in color) also contained higher nitrite and nitrate levels compared to white house‐EBNs, suggesting a relationship between nitrite and nitrate with EBN's color. Of note, we detected no presence of hemoglobin in red “blood” nest. Using infrared spectra analysis, we demonstrated that red/brown cave‐EBNs contained higher intensities of C‐N and N‐O bonds compared to white house‐EBNs. Together, our study suggested that the color of EBNs was associated with the prevalence of the nitrite and nitrate contents.     

Quantification of Nitrite/Nitrate in Food Stuff Samples Using 2-Aminobenzoic Acid as a New Amine in Diazocoupling Reaction

2-Aminobenzoic acid has been used as an amine in diazocoupling reaction to form an azo dye in the quantification of nitrite/nitrate at trace level. The formed azo dye has an absorption maximum at 550 nm in aqueous phase, and the resulted dye can be extracted into organic solvent to lower the detection limit. The method obeys Beer's law in the concentration range 0–10 μg of nitrite in 25 ml of aqueous solution with a molar absorptivity of 3.6 × 10³ L mol⁻¹ cm⁻¹ and 0–2 μg of nitrite in 5 ml of organic phase. The detection limit of the dye has been found to be 0.056 μg ml⁻¹. Nitrate is determined by reducing it to nitrite after passing through a copperized cadmium reductor column. The effect of interfering ions on the determination of nitrite/nitrate has been described. The developed method has been applied to determine the nitrite/nitrate trace level in vegetable, fruit juice, and milk powder samples.     

Nitrate and nitrite quantification from cured meat and vegetables and their estimated dietary intake in Australians

High dietary nitrate and nitrite intake may increase the risk of gastro-intestinal cancers due to the in vivo formation of carcinogenic chemicals known as N-nitroso compounds. Water and leafy vegetables are natural sources of dietary nitrate, whereas cured meats are the major sources of dietary nitrite. This paper describes a simple and fast analytical method for determining nitrate and nitrite contents in vegetables and meat, using reversed-phase HPLC-UV. The linearity R² value was >0.998 for the anions. The limits of quantification for nitrite and nitrate were 5.0 and 2.5 mg/kg, respectively. This method is applicable for both leafy vegetable and meat samples. A range of vegetables was tested, which contained <23 mg/kg nitrite, but as much as 5000 mg/kg of nitrate. In cured and fresh meat samples, nitrate content ranged from 3.7 to 139.5 mg/kg, and nitrite content ranged from 3.7 to 86.7 mg/kg. These were below the regulatory limits set by food standards Australia and New Zealand (FSANZ). Based on the average consumption of these vegetables and cured meat in Australia, the estimated dietary intake for nitrate and nitrite for Australians were 267 and 5.3 mg/adult/day, respectively.     

The oral nitrate and nitrite intake in The Netherlands: evaluation of the results obtained by HPIC analysis of duplicate 24-hour diet samples collected in 1994.

In spring and autumn of 1994 duplicates of 24-h diets were collected from 123 respondents. One of the goals of this study was to determine the amount of nitrite and nitrate in the duplicates of 24-h diets to establish the oral daily intake of these analytes. For this purpose an HPIC/UV method for the determination of nitrate and nitrite in duplicate diets was developed and validated. The sample preparation procedure was derived from the in-house method used for the determination of nitrate and nitrite in human blood plasma. The sample is diluted with water, deproteinized with Carrez reagent, followed by chromatographic clean-up on an SPE C18-column. Both the nitrate and the nitrite results are quantitative. The recovery for nitrite was on average 104% (n = 21, spiking levels: 0.84-95 mg/kg) and for nitrate on average 103% (N = 21, spiking levels: 1.8-404 mg/kg). Samples of duplicates of 24-h diets were analysed according to the method developed. The median intake of nitrite calculated from the samples collected in spring 1994 was 0.6 mg/person day (range < 0.1-6.1 mg/person/day). For the samples collected in autumn 1994 these figures were < 0.2 mg/person/day (range < 0.1-16 mg/person/day). The mean intake of nitrate was 73 mg/person/day (range 7-322 mg/person/day) in spring 1994 and 87 mg/person/day (range 1-310 mg/person/day) in autumn 1994. The overall mean intake of nitrate in 1994 was 80 mg/person/day. The daily intake for nitrate was higher than that found in the duplicate diet study carried out in 1984/1985, when an average daily intake of 52 mg/person was measured. The intake of nitrite was also higher than found in the duplicate diets collected in 1984/1985. The findings of the study are discussed in the context of the ADI for nitrate and nitrite as well as the outcome of other recent European intake studies.     

The influence of nitrite and nitrate on microbial, chemical and sensory parameters of slow dry fermented sausage

Nitrate and/or nitrite are used in the manufacture of dry-fermented sausages. However, research has mainly been focusing on nitrite and its effect on flavour development whereas little attention has been paid to nitrate. The aim of the present work was to study the effect of nitrate and nitrite as curing salts on the quality of a slow fermentation process. Two different batches containing nitrate or nitrite were manufactured. Microbial and chemical parameters were monitored during ripening and after vacuum packed storage, as well as their fatty acid composition and their profile of volatile compounds. The oxidation, measured as TBARS (Thiobarbituric acid reactive substances), was greater in the samples with added nitrite than in the samples with added nitrate. FFA (free fatty acids) release was higher in the samples containing nitrite throughout the process. Volatile compounds arising from amino acid degradation and carbohydrate fermentation were generated at higher levels in the samples with added nitrate, probably due to the higher population of microorganisms in these samples and the effect of nitrate on their metabolism.     

Effect of nitrate/nitrite on the quality of sausage (sucuk) during ripening and storage

The effects of nitrate/nitrite on the microbial and chemical properties and sensory quality of Turkish‐style sausage (sucuk) were investigated during 15 days of ripening and 45 days of storage. Aerobic plate count, mould and yeast count, pH, 2‐thiobarbituric acid value, residual nitrite level, nitrosomyoglobin conversion and sensory characteristics (flavour, colour and cutting scores) were monitored. Aerobic plate count increased (P < 0.05) during the first 8 days of ripening and decreased (P < 0.05) during further ripening and storage. Mould and yeast count increased (P < 0.05) during the first 4 days of ripening and decreased (P < 0.05) during further ripening and storage. Overall sensory quality increased (P < 0.05) during the first 12 days of ripening and decreased (P < 0.05) during further ripening and storage. Increasing the nitrate/nitrite concentration increased (P < 0.05) the overall sensory quality. During the first 4 days of ripening, the pH of all sausages decreased (P < 0.05) from 5.98 to 4.53–4.81, owing to the action of lactic acid bacteria. Residual nitrite level decreased (P < 0.05) sharply during the first 8 days of ripening, from 150 to about 2 mg kg^?1 in sausage samples B3 (prepared with 150 mg kg^?1 nitrite, 300 mg kg^?1 nitrate and starter culture) and from 75 to about 1 mg kg^?1 in samples B2 (prepared with 75 mg kg^?1 nitrite, 150 mg kg^?1 nitrate and starter culture). The conversion of haem pigments to nitrosomyoglobin increased (P < 0.05) during the first 12 days of ripening and decreased (P < 0.05) during further ripening and storage. Copyright © 2004 Society of Chemical Industry     

Nitrate and nitrite in vegetables from areas affected by war-time operations in Croatia

To evaluate nitrate and nitrite contents in cabbage and potatoes, the most consumed vegetables in Croatia, and to examine if war-time operations in Croatia affected nitrate and nitrite levels in vegetables, potatoes and cabbage cultivated in the war region (East Slavonia) and out of war regions were analyzed. Data showed that nitrate contents were higher in the samples from the war region (32% and 24% in potatoes and cabbage, respectively) but differences were not statistically significant. The nitrate levels found in potato samples (196 mg NaNO₃/kg fresh weight (FW) and cabbage samples (911 mg NaNO₃/kg FW) were comparable with levels reported for other countries. The nitrite contents in potatoes (321 μg NaNO₂/kg FW) and cabbage (173 μg NaNO₂/kg FW) were lower than the contents reported for most other countries. Further examinations of nitrate and nitrite concentrations in vegetables in Croatia as well as examinations of the influence of war-time operations on accumulation of these toxic contaminants are necessary.