A flow-injection spectrophotometric method for nitrate and nitrite determination through nitric oxide generation

A flow injection method with novel spectrophotometric detection for the determination of nitrite and nitrate in foodstuffs is presented. The method is based on the reduction of nitrite and nitrate to nitric oxide, with subsequent reaction with iron (II) and thiocyanate in an acid medium, forming FeSCNNO⁺. The absorbance of the complex, with a maximum at 460 nm, is proportional to the nitrite and nitrate concentrations. The NO is generated in two stages: (1) reduction of nitrate to nitrite in a cadmium copper reductor column and (2) reduction of the nitrite to NO in a sulfuric acid medium. The influence of reagent concentrations and manifold parameters were evaluated. Nitrite and nitrate can be determined in the range of 0.30–3.00 and 1.00–10.00 mg l⁻¹, respectively. The sampling rate of analyses was 30–40 h⁻¹ and, considering a sample of 5.0 g, the determination limit of the method was 20 and 13 mg kg⁻¹ of nitrate and nitrite, respectively. Nitrite and nitrate were determined in vegetables and meat products by the proposed method. The precision and accuracy of the proposed method were comparable to those of the reference spectrophotometric method (official AOAC reference method for the determination of nitrate in foodstuffs).     

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.     

Modification of the ion exchange HPLC procedure for the detection of nitrate and nitrite in dairy products

Existing ion exchange HPLC methodology for nitrate and nitrite analysis in cured meat products suffers from high analyte variability at low concentrations and also chromatographic interference by artifacts in some other foods, such as dairy products. An investigation into the sources of variability has shown that both the cyclohexyl solid phase extraction cartridge and the glass fibre filter used in the original method can introduce artifacts which interfere with the determination of the nitrate in foodstuffs. We have also found that the use of a graphitized solid phase extraction cartridge used in tandem with the cyclohexyl solid phase extraction cartridge removed the artifacts from the chromatograph of dairy products that co-eluted with nitrite and nitrate. Values for the nitrite and nitrate content of dairy products were obtained by the HPLC procedure using these two solid phase extraction cartridges and the values obtained were in close agreement with those obtained by cadmium column reduction and colorimetry.     

Nitrate and nitrite in vegetables on the Danish market: content and intake

The contents of nitrate and nitrite in lettuce, leek, potato, beetroot, Chinese cabbage and white cabbage on the Danish market were determined for 3 years in the period 1993-1997 as part of the Danish food monitoring programme. These vegetables are supposed to provide the major contribution to the intake of nitrate from the diet. Results for nitrate and nitrite in fresh and frozen spinach are also shown. The highest content of nitrate was found in lettuce followed by beetroot, Chinese cabbage, fresh spinach, leek, frozen spinach, white cabbage and potatoes. For all the products a great variation in the content of nitrate was found. For lettuce a characteristic variation throughout the year is clearly seen with the highest content in the winter period the lowest content in the summer period. Generally, the content of nitrite was low but in spinach high contents were found, probably due to improper storage conditions during transportation. The intake of nitrate and nitrite from these vegetables is calculated on the basis of two different consumption surveys. For both surveys the average intake of nitrate from the vegetables included in the monitoring programme is estimated to be approximately 40mg day^-1, whereas for nitrite the average intake is approximately 0.09mg day^-1. The total intake of nitrate and nitrite is estimated to be respectively 61mg day^-1 and 0.5mg day^-1.     

Nitrate and nitrite in vegetables from north China: content and intake

The contents of nitrate and nitrite in potato, cabbage, Chinese cabbage, scallion (shallot), celery, cucumber, tomato, eggplant and wax gourd taken from the north China market from 1998 to 1999 were determined. These vegetables provide the major contribution to the nitrate intake from the diet. The highest content of nitrate was found in celery followed by Chinese cabbage, cabbage, scallion, wax gourd and eggplant. For all the products, a great variation in the content of nitrate was found. Generally, the nitrite content was low. The average intake of nitrate and nitrite from these vegetables was estimated as approximately 422.8 and 0.68 mg day -1 , respectively.     

Residue analysis of nitric oxide fumigation in nine stored grain and nut products

Nitric oxide (NO) is a recently discovered fumigant for postharvest pest control on fresh and stored products. Nitric oxide fumigation also does not leave residues on fresh fruit and vegetables when conducted properly. In this study, we analyzed nitrate (NO₃⁻) and nitrite (NO₂⁻) levels in liquid extracts and nitrogen dioxide (NO₂) desorption rates as residues of NO fumigation at various times after fumigation on nine stored grain and nut products. Each product was fumigated separately with 3.0% NO for 24 h in two treatments: one treatment (NO–N₂) was terminated with nitrogen gas (N₂) flush and the other (NO-Air) was terminated with normal air flush. For NO–N₂, NO₃⁻ concentrations of all fumigated products were not significantly higher than those of untreated controls at 1, 7, and 14 d after fumigation. NO₂⁻ concentrations of all fumigated products from N₂ gas flush were not significantly higher than those of control products at 14 d after fumigation. NO₂ desorption rates for most products from NO–N₂ treatment showed no significant difference from those for the controls 1 d after NO fumigation, except for beans and wheat, which showed no significant difference at ≥7 d after fumigation. All products from NO-Air treatment, however, had significant higher NO₃⁻ and NO₂⁻ ion concentrations in liquid extracts at 14 d after fumigation than those from NO–N₂ treatment and the control. NO₂ desorption rates in all products from NO-Air treatment were also significantly higher than those from NO–N₂ treatment and the control at 21 d after fumigation. Therefore, when terminated properly with N₂ flush, NO fumigation did not result in significant increases of NO₃⁻, NO₂⁻, or NO₂ as residue in nut and grain products.     

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：     

Dietary exposures to nitrate in the UK

Dietary exposures to nitrate have been estimated for two different groups of UK consumers. The daily dietary exposure of the general population was estimated from the UK 1997 Total Diet study to be 52mg/ day. Vegetables contributed approximately 70% to this total dietary exposure. Dietary exposure estimates for adult consumers of vegetables commonly eaten in the UK were 93mg/day and 140mg/day for mean and 97.5th percentile consumers, respectively. Dietary exposures to nitrate of UK consumers are similar to those in other European countries and are less than the Acceptable Daily Intake (ADI) for nitrate set by the European Commission's Scientific Committee for Food.     

Nutrition and sarcopenia: A review of the evidence of nutritional influences

Prevention of age related decline in muscle mass and strength is a key strategy to keeping physical capacity in older age and allowing independent living. To emerge preventive strategies, a better understanding is required of life style factors that impacts on sarcopenia. However, since muscle mass and strength in later life depend on both the rate of muscle loss and the peak achieved in early life, attempts to prevent sarcopenia also require considering diet through the life course and the potential benefits of early interventions. Optimizing diet and nutrition status during the life may be an important strategy to preventing sarcopenia and enhancing physical ability in older age.     

Metabolism of nitrate in fermented meats: the characteristic feature of a specific group of fermented foods

Within the universe of food fermentation processes the multi-purpose use of nitrate and/or nitrite is a unique characteristic of meat fermentations. These curing agents play a decisive role in obtaining the specific sensory properties, stability and hygienic safety of products such as fermented sausages, ham and, more recently, emulsion type of sausages. The use of nitrate is the traditional method in curing processes and requires its reduction to reactive nitrite. Thus, nitrate reduction is the key event that is exclusively performed by microorganisms. Under controlled fermentation conditions starter cultures are used that contain staphylococci and/or Kocuria varians, which in addition to strongly affecting sensory properties exhibit efficient nitrate reductase activity. To obtain clean label products some plant sources of nitrate have been in use. When producing thermally treated sausages (e.g. of emulsion type), starter cultures are used that form nitrite before cooking takes place. Staphylococci reduce nitrite to ammonia after nitrate has been consumed. K. varians is devoid of nitrite reductase activity. Nitrate and nitrite reductases are also present in certain strains of lactobacilli. It was shown that their application as starter cultures warrants efficient activity in sausages made with either nitrate or nitrite. NO is formed from nitrite in numerous chemical reactions among which disproportionation and reaction with reductants either added or endogenous in meat are of practical importance. Numerous nitrosation and nitrosylation reactions take place in the meat matrix among which the formation of nitrosomyoglobin is of major sensory importance.Safety considerations in meat fermentation relate to the safe nature of the starter organisms and to the use of nitrate/nitrite. Staphylococci (“micrococci”) in fermented meat have a long tradition in food use but have not received the QPS status from the EFSA. They require, therefore, thorough assessment with regard to toxigenicity and pathogenicity determinants as well as presence of transferable antibiotic resistance. Nitrate and nitrite are still considered basically undesired in food. The main objections are based on their potential to form nitrosamines with carcinogenic potential. In view of new results from intensive research of NO, potential risks are opposed by positive effects on human health.     

试论中餐餐馆饮食存在的营养问题与对策

本文综述了中餐餐馆饮食存在的营养方面的问题,并对如何改进中餐餐馆饮食,做到合理营养,保证就餐者的身体健康提出了建议。     

Changes in nitrate and nitrite concentrations over 24 h for sweet basil and scallions

Nitrate and nitrite concentrations were determined for sweet basil and scallions over 24 h to determine if time of sampling or harvest impacts concentrations in raw vegetables. Also, nitrate and nitrite concentrations were determined separately for various edible parts of these plants. Basil had significant changes in nitrate and nitrite concentrations over a 24 h period. Nitrate was correlated to changes in light intensity with a 3 h lag time. The highest nitrate concentrations in basil (2777 ppm) occurred around 3 h after the light intensity peaked and had low values (165–574 ppm) during the dark period. The scallion nitrate and nitrite concentrations were always low but nitrate showed a peak a few hours before sunrise. Nitrate and nitrite concentrations in some raw vegetables may be reduced by harvesting at the best time of day for each type of plant. Nitrate concentrations were different in the edible plant parts tested.     

Effect of nitrate and nitrite curing salts on the generation and oxidation of fatty acids in non-fermented sausages

 The purpose of this work was to study the effect of nitrate and nitrite curing salts on FFA generation and oxidation during an industrial process of country-style dry sausages (35/40 mm casing diameter) to determine their role in flavour development. The pH values of the sausages are around 6.0 during the dry processing. FFA liberation is two or three times greater in the samples with nitrite than in those with nitrate. However, the intensity of the oxidative phenomena during the drying process does not depend on the quantity of FFA. In the samples with nitrate more intense oxidative phenomena take place, which lead to a stronger typical dry-cured flavour than in samples with nitrite. Received: 14 July 2000 / Revised version: 15 September 2000     

Evolution of nitrate and nitrite during the processing of dry-cured ham with partial replacement of NaCl by other chloride salts

Nitrate and nitrite are commonly added to dry-cured ham to provide protection against pathogen microorganisms, especially Clostridium botulinum. Both nitrate and nitrite were monitored with ion chromatography in dry-cured hams salted with different NaCl formulations (NaCl partially replaced by KCl and/or CaCl(2), and MgCl(2)). Nitrate, that is more stable than nitrite, diffuses into the ham and acts as a reservoir for nitrite generation. A correct nitrate and nitrite penetration was detected from the surface to the inner zones of the hams throughout its processing, independently of the salt formulation. Nitrate and nitrite achieved similar concentrations, around 37 and 2.2 ppm, respectively in the inner zones of the ham for the three assayed salt formulations at the end of the process, which are in compliance with European regulations.     

Arginine and nitric oxide synthase: Regulatory mechanisms and cardiovascular aspects

l ‐Arginine (l ‐Arg) is a conditionally essential amino acid in the human diet. The most common dietary sources of l ‐Arg are meat, poultry and fish. l ‐Arg is the precursor for the synthesis of nitric oxide (NO); a key signaling molecule via NO synthase (NOS). Endogenous NOS inhibitors such as asymmetric‐dimethyl‐l ‐Arg inhibit NO synthesis in vivo by competing with l ‐Arg at the active site of NOS. In addition, NOS possesses the ability to be “uncoupled” to produce superoxide anion instead of NO. Reduced NO bioavailability may play an essential role in cardiovascular pathologies and metabolic diseases. l ‐Arg deficiency syndromes in humans involve endothelial inflammation and immune dysfunctions. Exogenous administration of l ‐Arg restores NO bioavailability, but it has not been possible to demonstrate, that l ‐Arg supplementation improved endothelial function in cardiovascular disease such as heart failure or hypertension. l ‐Arg supplementation may be a novel therapy for obesity and metabolic syndrome. The utility of l ‐Arg supplementation in the treatment of l ‐Arg deficiency syndromes remains to be established. Clinical trials need to continue to determine the optimal concentrations and combinations of l ‐Arg, with other protective compounds such as tetrahydrobiopterin (BH₄), and antioxidants to combat oxidative stress that drives down NO production in humans.     

Ascorbic acid,nitrate, and nitrite concentration relationship to the 24 hour light/dark cycle for spinach grown in different conditions

Nitrate, nitrite and ascorbic acid (vitamin C) concentrations were determined for spinach (Spinacia oleracea L.) over a 24 h period to determine if light intensity (including dark periods) at time of harvest impacts concentrations in raw vegetables. Nitrate, nitrate and ascorbic acid concentrations varied significantly over the 24 h period and appeared to be related to changes in light intensity. Light intensity at the time an experimental sample is collected may affect the concentration of some constituents that a researcher is studying. Also, nitrate and nitrite concentrations in raw spinach can be reduced by harvesting at the best time of day. The highest nitrate concentrations in spinach occurred in the dark just prior to an increase in light intensity. Ascorbic acid was near its highest level for the 24 h period when the light intensity initially increased, then decreased to its lowest level around 3–6 h later.     

Survival of Listeria innocua in dry fermented sausages and changes in the typical microbiota and volatile profile as affected by the concentration of nitrate and nitrite

The involvement of nitrate and nitrite in the formation of N-nitrosamines in foods is a matter of great concern. This situation has led to revise the real amount of nitrate and nitrite needed in meat products to exert proper technological and safety activities, and also to extensive research to find alternatives to their use. The present study addresses the possibility of reducing the ingoing amounts of these additives below the legal limits established by the current European regulations. Different concentrations of nitrate and nitrite were tested on Spanish salchichón-type dry fermented sausages concerning their role in the microbiota and volatile profile. Sausages were manufactured with the maximum ingoing amounts established by the EU regulations (150 ppm NaNO₃ and 150 ppm NaNO₂), a 25% reduction and a 50% reduction; control sausages with no nitrate/nitrite addition were also prepared. The mixtures were inoculated with 5 log cfu/g of Listeria innocua as a surrogate for Listeria monocytogenes. L. innocua numbers in the final product were approximately 1.5 log cfu/g lower in the batch with the maximum nitrate/nitrite concentration when compared to 25 and 50% reduced batches, and about 2 log cfu/g in comparison to the control sausages. The final numbers of catalase-positive cocci were 1 log cfu/g higher in the 50% nitrate/nitrite reduced batch and 2 log cfu/g higher in the control sausages, compared to products manufactured with the maximum nitrate/nitrite concentration. This increase was related to a higher amount of volatile compounds derived from carbohydrate fermentation and amino acid degradation. Sausages with no addition of nitrate/nitrite showed higher amount of volatiles from lipid oxidation. Enterobacteriaceae counts reached detectable values (1-2 log cfu/g) in both nitrate/nitrite reduced sausages and in the control batch, while these organisms were not detected in the batch with the maximum ingoing amount. Nitrate and nitrite exerted a significant effect on the typical microbiota of dry fermented sausages and effectively contributed to control Listeria. These considerations should be taken into account in view of a future restriction in the use of these curing additives.     

不同时间一氧化氮(NO)熏蒸处理对番茄采后品质的影响

以跃变性果蔬番茄为试材,观测了20μL/L NO气体处理0.5、1、1.5、2 h对番茄采后失质量、呼吸强度、果肉硬度及可溶性固形物的影响.结果表明:在室温条件下,20μL/L的外源NO处理番茄果实1～2 h能够有效控制果实的采后质量损失、果肉硬度下降.调控果实呼吸变化,控制果实可溶性固形物的变化,说明NO处理能够延缓番茄果实的采后成熟及衰老过程;仅处理0.5 h的处理对番茄的采后品质无显著作用.