Nitrate and Nitrite Methods of Analysis and Levels in Raw Carrots, Processed Carrots and in Selected Vegetables and Grain Products

Methods for quantitative estimation of nitrate and nitrite were compared. Levels of these ions were measured in vegetables and grain products and effects of processing on nitrate and nitrite levels in carrots were measured. These data allow more accurate estimation of ingestion levels and suggest means to reduce exposure to these ions. High performance liquid chromatography (HPLC) had better precision and recoveries than either a classical Cd-Griess method nitrate or a Griess method for nitrite. Nitrate concentration by HPLC varied greatly within and between vegetables, ranging from 1 μmol/100g in mushrooms to 5000 μmol/100g in celery and averaging 9.7 ± 4.4 μmol/100g in grains. Nitrate levels in vegetables sold as “organic” were not different (p<0.05) from conventional vegetables. No nitrite was detected in either vegetables or grains. Nitrate was unevenly distributed in carrots with the core having the most. Storage of carrots at -18°C for 10 wk did not alter nitrate levels and no nitrite developed. Fifty-seven percent of nitrate was leached into cooking liquid when frozen carrots were boiled. Thirty-two percent of nitrate was lost during canning and 47% of the remainder was in the liquid. No nitrite developed during 10 wk of canned storage.     

Prediction of NaCl,nitrate and nitrite contents in minced meat by using a voltammetric electronic tongue and an impedimetric sensor

A method for predicting levels of sodium chloride, sodium nitrite and potassium nitrate in minced meat by using a combination of two different electrochemical methods; namely an electronic tongue (ET) based on pulse voltammetry and electrochemical impedance spectroscopy measurements is proposed here. The measurements with the voltammetric ET were carried out on both saline solutions (brines) and minced meat, whereas the impedimetric sensor was used only in minced meat. The addition of the salts was performed following an experimental design in which a system of three compounds/three levels was established. Multivariate analysis including Cross validation and Partial Least Square (PLS) techniques were applied for data management and prediction models building. A very good prediction of the concentration of chloride was achieved, whereas the prediction for the concentration of nitrate and nitrite can be considered as moderate.     

冷藏对蔬菜中亚硝酸盐及硝酸盐含量的影响

目的 探索冰箱冷藏(4℃)条件下蔬菜中硝酸盐和亚硝酸盐含量随时间的变化,对冷藏食品的安全性进行评价.方法 分别采用重氮偶合分光光度法和麝香草酚分光光度法对生菜、菠菜、油麦菜及苦苣4种常用蔬菜在冷藏条件下其亚硝酸盐及硝酸盐的含量变化进行测定.结果 1～4d内,生菜、苦苣中的亚硝酸盐含量随着时间的延长逐渐增加;菠菜、油麦菜中亚硝酸盐含量分别在第2天和第3天达到最大,之后逐渐降低,而菠菜中亚硝酸盐含量在第4天又出现上升趋势.4种蔬菜中亚硝酸盐含量4d中最大值仅为0.419 4 mg/kg(生菜),均＜4 mg/kg的限量值.苦苣和油麦菜中硝酸盐含量在1～4d内逐渐上升.菠菜中硝酸盐含量第2天达到418.48 mg/kg,之后又降低至20.83 mg/kg.生菜中硝酸盐含量在第3天达到最大值317.26 mg/kg,之后逐渐下降,但均在安全范围内(432 mg/kg).结论 在4℃下冷藏4d后,4种常见蔬菜均可安全食用.可见,冷藏能有效减缓亚硝酸盐和硝酸盐含量的升高.     

Time-dependent depletion of nitrite in pork/beef and chicken meat products and its effect on nitrite intake estimation

The food additive nitrite (E249, E250) is commonly used in meat curing as a food preservation method. Because of potential negative health effects of nitrite, its use is strictly regulated. In an earlier study we have shown that the calculated intake of nitrite in children can exceed the acceptable daily intake (ADI) when conversion from dietary nitrate to nitrite is included. This study examined time-dependent changes in nitrite levels in four Swedish meat products frequently eaten by children: pork/beef sausage, liver paté and two types of chicken sausage, and how the production process, storage and also boiling (e.g., simmering in salted water) and frying affect the initial added nitrite level. The results showed a steep decrease in nitrite level between the point of addition to the product and the first sampling of the product 24 h later. After this time, residual nitrite levels continued to decrease, but much more slowly, until the recommended use-by date. Interestingly, this continuing decrease in nitrite was much smaller in the chicken products than in the pork/beef products. In a pilot study on pork/beef sausage, we found no effects of boiling on residual nitrite levels, but frying decreased nitrite levels by 50%. In scenarios of time-dependent depletion of nitrite using the data obtained for sausages to represent all cured meat products and including conversion from dietary nitrate, calculated nitrite intake in 4-year-old children generally exceeded the ADI. Moreover, the actual intake of nitrite from cured meat is dependent on the type of meat source, with a higher residual nitrite levels in chicken products compared with pork/beef products. This may result in increased nitrite exposure among consumers shifting their consumption pattern of processed meats from red to white meat products.     

Intake and risk assessment of nitrate and nitrite from New Zealand foods and drinking water

Exposure to excess nitrite is a potential health risk for humans. One hundred meat and processed foods and 100 vegetable samples purchased from New Zealand retail outlets were prepared as for consumption and analysed for nitrite and nitrate concentration using a standard, validated methodology. Nitrate concentrations ranged from less than the limit of detection (LOD = 5 mg kg^-1) in cheddar cheese and cream cheese-based dips to 3420 mg kg^-1 in lettuce. Nitrite was detected in half the processed foods and meats analysed (levels up to 119 mg kg^-1), but detected in only one vegetable sample above the LOD (broccoli at 27 mg kg^-1 nitrite). Concentration data were combined with 24 h dietary recall information to generate 4398 individual adult daily exposure scenarios for exogenous nitrite and nitrate including a contribution from water assessed from 1021 drinking water samples. The mean adult daily intake of exogenous nitrate and nitrite from food and water combined was 16 and 13% of the Acceptable Daily Intake (ADI), respectively, and therefore should not pose a health risk for the average consumer. A maximally exposed New Zealand adult is estimated to have an intake of up to seven times the ADI for nitrate. When the endogenous conversion of nitrate to nitrite is taken into account, approximately 10% of people with an average rate of conversion and half of all people with a high rate of conversion are estimated to exceed the ADI. Either the ADI is inappropriate and needs to be re-evaluated, or those individuals who have a high rate of conversion of nitrate to nitrite are at risk to adverse effects of nitrite exposure.     

Dried sausages fermented with Staphylococcus xylosus at different temperatures and with different ingredient levels - Part I. Chemical and bacteriological data

Sausages with added Staphylococcus xylosus were fermented at different temperatures and with different added levels of salt, glucose, nitrite, nitrate and Pediococcus pentosaceus in accordance with a six factor fractional design. The numbers of surviving Staphylococcus xylosus, lactic acid bacteria, pH, free fatty acids and residual amounts of nitrite and nitrate were measured. The effects of temperature and different ingredients on the chemical and bacterial data were tested using multiple linear regression and analysis of variance. The study showed that numbers of surviving Staphylococcus xylosus were reduced by high fermentation temperature, especially when salt concentration was low and glucose concentration high. High levels of nitrite and addition of Pediococcus pentosaceus had adverse effects as well. pH was lowered by high fermentation temperature, low salt concentration and addition of glucose and Pediococcus pentosaceus. On the other hand p H was increased by addition of nitrate. The pH-lowering effect of glucose was small when temperature was low. The residual levels of nitrite and nitrate were increased by addition of nitrate, but then increased and decreased, respectively, by increasing temperature. Addition of glucose and Pedio coccus pentosaceus lowered the residual amount of nitrite. The level of free fatty acids was reduced by salt and increased by higher temperature. Of the individual acids, the amount of linoleic acid was also increased by addition of Pediococcus pentosaceus and decreased by nitrate.     

离子色谱法测定牛奶或奶粉中亚硝酸盐 硝酸盐及氯化物

牛奶或奶粉中的亚硝酸盐、硝酸盐及氯化物经溶解稀释后可直接进样至离子色谱仪中进行测定；样品中加入亚硝酸盐、硝酸盐及氯化物标准后测定，两者的准确度及精密度均取得满意结果。6份样品采用本方法测定结果与国家标准[GB／T5413．32-1997及GB／T5413．32-1997（第二法）]采用的方法测定结果比较无显著性差异：     

Monitoring nitrite and nitrate residues in frankfurters during processing and storage

Frankfurter-type sausages were prepared in a pilot plant with different concentrations of NaNO(2) (75, 125 or 250 ppm) combined or not with 200 ppm KNO(3). A meat system, free of curing agents, was also used as control. Nitrite and nitrate levels were tested in various processing steps and over 120 days storage at 3 °C of the vacuum-packaged frankfurters. Little influence of the originally added nitrite level on the amount of nitrate formed was observed. Important losses of nitrite and nitrate were due to cooking. Thereafter about 50% of the nitrite added initially remained in this form in all samples (39, 59 and 146 ppm, respectively) and between 10 and 15% as nitrate. When only nitrate was initially added, formation of nitrite after cooking was observed (maximum level 43 ppm NaNO(2)). Formulations prepared with both nitrate and nitrite showed no significant differences (p < 0.01) respect to their nitrite or nitrate counterparts. A good correlation among nitrite and nitrate levels and storage time was showed by multiple linear regression analysis. It is concluded that the use of nitrate in combination with nitrite in cooked meat products seems to have little technological significance and adds to the total body burden of nitrite.     

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.     

Changes in nitrate and nitrite content of four vegetables during storage at refrigerated and ambient temperatures

The nitrate and nitrite contents of four kinds of vegetables (spinach, crown daisy, organic Chinese spinach and organic non-heading Chinese cabbage) in Taiwan were determined during storage at both refrigerated (5 ± 1°C) and ambient temperatures (22 ± 1°C) for 7 days. During storage at ambient temperature, nitrate levels in the vegetables dropped significantly from the third day while nitrite levels increased dramatically from the fourth day of storage. However, refrigerated storage did not lead to changes in nitrate and nitrite levels in the vegetables over 7 days.     

Exposure estimates of nitrite and nitrate from consumption of cured meat products by the U.S. population

The dietary exposures of nitrite and nitrate from consumption of cured meat products were estimated for the U.S. population aged 2 years and older, and children aged 2 to 5 years, using both 2-day food consumption data from the publicly available combined 2009–2012 National Health and Nutrition Examination Survey (NHANES) and 10–14-day food consumption data from the 2009 and 2012 NPD Group, Inc. National Eating Trends-Nutrient Intake database (NPD NET-NID), and residual nitrite and nitrate levels in cured meat products available from the recent American Meat Institute Foundation/National Pork Board (AMIF/NPB) national market survey of the nitrite and nitrate levels in cured meat products in the U.S.A. The dietary exposure for consumers of cured meat products (eaters-only) was estimated at the mean and 90th percentile for three exposure scenarios: low exposure, average exposure, and high exposure, to account for the range in the amount of nitrite and nitrate in a given cured meat product category. In addition, a cumulative exposure that takes into account all cured meat product categories containing nitrite and nitrate was determined, and the relative percent contribution of each cured meat product category to the cumulative exposure was estimated. Cured, cooked sausages and whole-muscle brine-cured products were the two major contributing categories to dietary exposure of nitrite and nitrate for both U.S. population aged 2 years and older and children aged 2–5 years.     

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.     

离子色谱法测定FAPAS比对肉粉样品中亚硝酸盐、硝酸盐含量

为建立离子色谱法测定肉粉样品中亚硝酸盐和硝酸盐含量的方法,将样品经超声和水浴提取后,经阳离子 交换树脂柱净化,以氢氧化钾为淋洗液,经Dionex IonPacTMAS19阴离子柱分离,采用AERS 500（4 mm）阴离子抑 制器、电导检测器进行检测,标准曲线定量。结果表明：亚硝酸根离子在质量浓度0.10～4.00 mg/L、硝酸根离子在 质量浓度0.50～20.00 mg/L范围内线性关系良好,相关系数均为0.999 9;样品中亚硝酸盐和硝酸盐的平均回收率分 别为99.53%（n=9）和99.29%（n=9）,相对标准偏差分别为0.60%和0.25%。本次比对结果为“满意”,表明本方 法科学、可靠,适用于基质复杂的熟肉制品中亚硝酸盐和硝酸盐的测定。     

Development and Validation of a Method for Determination of Residual Nitrite/Nitrate in Foodstuffs and Water After Zinc Reduction

An environmentally friendly and cost-effective spectrophotometric method to analyze nitrate and/or nitrite was developed. The method is based on reduction of nitrate with zinc powder (instead of the cadmium or enzymes used in the standard methods approved by ISO/CEN). The initial nitrite concentration and total nitrite after reduction are determined by the very sensitive and widely used diazotization-coupling Griess reaction. A single-laboratory validation was applied in five different matrices (vegetable, meat product, baby food, dairy product, and surface water). The results show that the new method fulfills the international criteria for precision and recovery. The limit of detection for several matrices, calculated using fortified samples, ranged from 3 to 5 mg/kg for both nitrite and nitrate. Furthermore, the results obtained were in good agreement with those obtained using the CEN method (HPLC) and the ISO method (Cd reduction).     

连续流动分析-盐酸萘乙二胺分光光度法测定酱油中的亚硝酸盐和硝酸盐

建立了一种同时测定酱油中亚硝酸盐和硝酸盐的连续流动分析-盐酸萘乙二胺分光光度法。酱油样品经25 ℃超声15 min预处理,乙酸锌-亚铁氰化钾体系沉淀蛋白质后,自动进样且采用气泡隔断样品,并经透析膜透析,采用双通道同时测量亚硝酸盐和硝酸盐。结果表明,亚硝酸盐在0.025～1.0 mg/L质量浓度范围内线性良好,加标回收率在83.54%～98.56%之间,精密度试验结果相对标准偏差（RSD）＜3%;硝酸盐在0.25～5.0 mg/L质量浓度范围内线性良好,加标回收率在92.95%～102.32%之间,精密度试验结果RSD＜2%。该方法准确、高效、灵敏,可以用于测定酱油中亚硝酸盐和硝酸盐。     

Nitric oxide inhibits the formation of zinc protoporphyrin IX and protoporphyrin IX

The aim of this study was to elucidate the mechanism by which curing agents, especially nitrite, inhibit the formation of zinc protoporphyrin IX (ZPP) in dry-cured hams such as Parma ham. The oxidation–reduction potential of model solutions was increased by the addition of nitrite, but it was not clear whether the formation of ZPP is inhibited by the oxidizing property of nitrite. The effect of nitric oxide (NO) produced from nitrite on the formation of ZPP was examined. The amount of ZPP formed was decreased by the addition of NO donors. The amount of protoporphyrin IX (PPIX), which is the precursor of ZPP, was also decreased by the addition of NO donors. It is concluded that NO produced from nitrite inhibited the formation of PPIX and ZPP was therefore not formed in cured meat products with the addition of nitrite or nitrate.     

Dried Pork as Influenced by Nitrate, Packaging Method and Storage

A new technique was developed to produce high quality, uniform and attractive thin sliced, cured dried pork. The effects of nitrate, packaging method and storage time on residual nitrite, TBA values, sensory properties and microbiological counts were determined. Residual nitrite decreased with increased storage time at 3 ± 1°C. The addition of nitrate plus vacuum packaging caused a greater residual nitrite level and a lower TBA value during storage. Nitrite and/or nitrate acted as an antioxidant to retard oxidative rancidity (TBA value). Dried pork manufactured by the technique described had no major rancidity problem and had an acceptable shelf life. Total aerobic plate counts, lactic acid producing microbial counts and total anaerobic counts were not affected by nitrate or packaging methods. Coliforms, molds and yeasts were not found in this dried pork.     

紫外检测离子色谱法测定酱腌菜中的亚硝酸盐与硝酸盐

建立了紫外检测离子色谱法测定酱腌菜中的亚硝酸盐与硝酸盐的分析方法。样品经超声提取后,以1.8 mmol/L Na2CO3+1.7 mmol/L NaHCO3为流动相,经Metrosep A Supp 4-250阴离子交换色谱柱分离,于210nm处紫外检测。结果表明:此法NO2-、NO3-的检出限分别为1.0 mg/kg和2.0 mg/kg,实际样品的加标回收率分别为80.4%~82.1%和93.1%~96.5%,相对标准偏差≤10%。     

鲜姜、煮姜、泡姜中Vc、硝酸盐、亚硝酸盐含量测定

测定姜经不同处理后Vc、硝酸盐和亚硝酸盐的含量，以评价不同处理方法对营养成分Vc和有害成分硝酸盐、亚硝酸盐的破坏情况。方法：通过紫外吸收分光光度计在特定波长下的吸收值来计算所测成分的舍量。结果：生姜经煮沸15min后，Vc的含量为2．30mg／100g，只有鲜姜中Vc的20．65％，而泡姜中Vc的含量达5．86mg／100g，为鲜姜中的52．60％：鲜姜中硝酸盐和亚硝酸盐的含量分别为39338．5mg／100g和0．3046mg／100g，煮姜和泡姜中硝酸盐和亚硝酸盐的含量都很低。