Analysis of four tobacco-specific nitrosamines in mainstream cigarette smoke of Virginia cigarettes by LC-MS/MS

An improved method was developed for the determination of the four major tobacco-specific nitrosamines（TSNAs） in mainstream cigarette smoke. The new method offers decreased sample preparation and analysis time as compared to traditional methods. This method uses isotope dilution liquid chromatography coupled with a tandem mass spectrometer with electrospray ionization and is significantly more sensitive than traditional methods. It also shows no evidence of artifactual formation of TSNAs. Sample concentrations were determined for four TSNAs in mainstream smoke using two isotopically labeled TSNAs analogues as internal standards. Mainstream smoke was collected on an industry standard 44-ram Cambridge filter pad, extracted with 0.1 mol/L ammonium acetate, purified by solid-phase extraction, and analyzed without further sample cleanup. The analytical column is a 3.9 mm×150 mm Waters Symmertry Shield RP18 column and volume fraction of the mobile phase is 50% methanol, 50% water containing 0.1% acetic acid. The results show that the linear range is 0.5-100.0 mg/L except for N-nitrosoanabasine （NAB） from 0.25 to 50.0 mg/L. The limits of detection are 0.1 mg/L for N-nitrosonornicotine （NNN）, 0.08 mg/L for 4-（methylnitrosamino）-1- （3-py-ridyl）-1-butanone （NNK）, 0.05 mg/L for N-nitrosoanatabine （NAT） and 0.06 mg/L for NAB. The recoveries of the four TSNAs are from 90.2% to 105.7%.

Hemoglobin Adducts as Biomarkers of Exposure to and metabolic Activation of Carcinogenic Tobacco-Specific Nitrosamines

The carcinogenic tobacco-specific nitrosamines N-nitrosonornicotine (NNN) and 4-(methylnitrosamino)-l-(3-pyridyl)-l-butanone (NNK) form hemoglobin adducts in laboratory animals and humans. These adducts release 4-hydroxy-l-(3-pyridyl)-l-butanone (HPB) upon mild base hydrolysis. HPB released from human hemoglobin can be quantified by gas chromatography-mass spectrometry. It is the only available biochemical marker for determination of exposure to, and metabolic activation of, carcinogens present only in tobacco. Levels of HPB were highest in snuff-dippers, followed by smokers and nonsmokers. Large interindividual variations in adduct levels were observed. The relationship between HPB levels in globin and DNA of rats treated with NNK has been investigated in order to aid in interpretation of the data from humans. These studies have provided the initial database for understanding the metabolic activation of tobacco-specific nitrosamines in humans.

Nitrosamines crisis in pharmaceuticals-Insights on toxicological implications,root causes and risk assessment:A systematic review

The presence of N-nitroso compounds,particularly N-nitrosamines,in pharmaceutical products has raised global safety concerns due to their significant genotoxic and mutagenic effects.This systematic review investigates their toxicity in active pharmaceutical ingredients(APIs),drug products,and pharmaceutical excipients,along with novel analytical strategies for detection,root cause analysis,reformulation strategies,and regulatory guidelines for nitrosamines.This review emphasizes the molecular toxicity of N-nitroso compounds,focusing on genotoxic,mutagenic,carcinogenic,and other physiological effects.Additionally,it addresses the ongoing nitrosamine crisis,the development of nitrosamine-free products,and the importance of sensitive detection methods and precise risk evaluation.This comprehensive overview will aid molecular biologists,analytical scientists,formulation scientists in research and development sector,and researchers involved in management of nitrosamine-induced toxicity and promoting safer pharmaceutical products.

Metabolism Study on Tobacco-specific N-Nitrosamines in Rabbit by Solid-phase Extraction and Liquid Chromatography-electrospray Ionization-mass Spectrometry Method

Metabolism of four tobacco-specific N-nitrosamines （TSNAs）, N＇-nitrosonornicotine （NNN）, N＇-nitrosoanatabine （NAT）, N＇-nitrosoanabasine （NAB） and 4-（methyl nitrosamino）-1- （3-pyridyl）-1-butanone （NNK） was studied by solid-phase extraction （SPE） and liquid chromato- graphy-electrospray ionization-mass spectrometry （LC-ESI-MS）. SPE and LC-ESI-MS method was evaluated to be rapid, simple, sensitive and selective for analysis of TSNAs in rabbit serum. The doses of TSNAs administrated were 4.666 μg/kg and 11.665 μg/kg according to different levels in cigarettes. Metabolic curves of four TSNAs and 4-（methyl nitrosamino）-1-（3-pyridyl）- 1-butanone （NNAL）, the metabolite of NNK, were obtained.

Association of a Tobacco-specific Nitrosamine Carcinogen with Urinary Cotinine,Urinary Sodium Excretion,and Total Energy Intake in Adolescents and Children

This study investigated the association of a tobacco-specific nitrosamine carcinogen,4-(methylnitrosamino)-1-(3-pyridy1)-I-butanol(NNAL)with urinary cotinine(uCot),urinary sodium(uNa)excretion,systolic blood pressure(sBP),and total energy intake in adolescents and children in relation to the subjects'age.A total of 790 subjects aged 6-19 years were evaluated.NNAL,uCot,corrected NNAL(cNNAL),the NNAL/uCot ratio,uNa,sBP,and nutrient intake were measured.A strong association between uCot and cNNAL was observed in children who were 11 years of age(r=0.881,P<0.001);however,no significant association was noted in adolescents who were 19 years of age.The uNa level was significantly higher(133.9 mmol/L vs.107.8 mmol/L,P<0.001)and sBP was significantly lower(105.3 mmHg vs.110.6 mmHg,P=0.012)in adolescents with elevated NNAL than in those without elevated NNAL.NNAL was significantly higher in subjects with increased uNa excretion than in those without increased uNa excretion.NNAL was positively correlated with uNa(r=0.183,P<0.001)and negatively correlated with sBP(r=0.142,P<0.001).Non-smokers with elevated NNAL/uCot ratios had significantly lower total energy intake than those without elevated NNAL/uCot ratios(1729.0 kcal/day vs.1911.0 kcal/day,P=0.008).The relationship between NNAL and uCot varied according to the subjects'age.NNAL seems to play a role in decreasing sBP by enhancing uNa excretion.Insufficient nutrient intake may contribute to endogenous formation of NNAL in non-smoking adolescents and children.

Intake of Volatile N-nitrosamines and Their Ability to Exogenously Synthesize in the Diet of Inhabitants from High-risk Area of Esophageal Cancer in Southern China

Objective Nan'ao County in Guandong Province is a high-risk area of esophageal cancer in Southern China. Of the suspected etiological factors in the environment, N-nitrosamines and their precursors have received the greatest attention. Methods Sixty samples of the diet ingested by the inhabitants were collected and detected for volatile N-nitrosamines and their precursors. Five N-nitrosamines detected by Gas Chromatography-Thermal Energy Analyzer were N-nitrosodimethylamine, N-nitrosodiethylamine, N-nitrosopyrrolidine, N-nitrosopiperidine and N-nitrosomethyl-benzylamine. Results The average content of 5 volatile N-nitrosamines in the diet was 312.0 μg/kg (median). The daily intake of the nitrosamines was 286.5 μg/head/day. Only the ability to exogenously synthesize N-nitrosopiperidine was powerful among 5 volatile N-nitrosamines. By a computerized stepwise regression analysis and curve fitting, we studied the correlation among the nitrosamines, the precursors and the major food items in the samples. Conclusion It demonstrated that a relatively high content of volatile N-nitrosamines was present in the diet collected in the area.

Nitrate-nitrite-nitrosamines exposure and the risk of type 1 diabetes: A review of current data

The potential toxic effects of nitrate-nitrite-nitrosamine on pancreatic β cell have remained a controversial issue over the past two decades. In this study, we reviewed epidemiological studies investigated the associations between nitrate-nitrite-nitrosamines exposure, from both diet and drinking water to ascertain whether these compounds may contribute to development of type 1 diabetes. To identify relevant studies, a systematic search strategy of Pub Med, Scopus, and Science Direct was conducted using queries including the key words "nitrate", "nitrite", "nitrosamine" with "type 1 diabetes" or "insulin dependent diabetes mellitus". All searches were limited to studies published in English. Ecologic surveys, case-control and cohort studies have indicated conflicting results in relation to nitrate-nitrite exposure from drinking water and the risk of type 1 diabetes. A null, sometimes even negative association has been mainly reported in regions with a mean nitrate levels < 25 mg/L in drinking water, while increased risk of type 1 diabetes was observed in those with a maximum nitrate levels > 40-80 mg/L. Limited data are available regarding the potential diabetogenic effect of nitrite from drinking water, although there is evidence indicating dietary nitrite could be a risk factor for development of type 1 diabetes, an effect however that seems to be significant in a higher range of acceptable limit for nitrate/nitrite. Current data regarding dietary exposure of nitrosamine and development of type 1 diabetes is also inconsistent. Considering to an increasing trend of type 1 diabetes mellitus(T1DM) along with an elevated nitrate-nitrite exposure, additional research is critical to clarify potential harmful effects of nitrate-nitritenitrosamine exposure on β-cell autoimmunity and the risk of T1DM.

Modifying NaY Zeolite with Metal Oxide by Microwave Irradiation:Influence on the Adsorption and Decomposition of N-nitrosamines

Coating MgO.br ZrO2 on zeolite NaY accelerated decomposition of N-nitrosodimethylamine and N-nitrosopyrrolidine, resulting from variation of the surface state such as generation of basic sites.

A New Automated Method and Sample Data Flow for Analysis of Volatile Nitrosamines in Human Urine

Volatile nitrosamines (VNAs) are a group of compounds classified as probable (group 2A) and possible (group 2B) carcinogens in humans. Along with certain foods and contaminated drinking water, VNAs are detected at high levels in tobacco products and in both mainstream and side-stream smoke. Our laboratory monitors six urinary VNAs—N-nitrosodimethylamine (NDMA), N-nitrosomethylethylamine (NMEA), N-nitrosodiethylamine (NDEA), N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), and N-nitrosomorpholine (NMOR)—using isotope dilution GC-MS/ MS (QQQ) for large population studies such as the National Health and Nutrition Examination Survey (NHANES). In this paper, we report for the first time a new automated sample preparation method to more efficiently quantitate these VNAs. Automation is done using Hamilton STARTM and Caliper StaccatoTM workstations. This new automated method reduces sample preparation time from 4 hours to 2.5 hours while maintaining precision (inter-run CV < 10%) and accuracy (85% - 111%). More importantly this method increases sample throughput while maintaining a low limit of detection (<10 pg/mL) for all analytes. A streamlined sample data flow was created in parallel to the automated method, in which samples can be tracked from receiving to final LIMs output with minimal human intervention, further minimizing human error in the sample preparation process. This new automated method and the sample data flow are currently applied in bio-monitoring of VNAs in the US non-institutionalized population NHANES 2013-2014 cycle.

Rapid and Simple Extraction Method for Volatile N-Nitrosamines in Meat Products

A new methodology for extraction, pre-concentration and analysis of volatile nitrosamines in meat-derived products was developed and compared with conventional methods (Distillation and two-step solid-phase extraction). The samples (canned sausages, cured meat, luncheon and smoked meat) were treated with an aqueous sodium hydroxide (NaOH) by autoclaving at 121°C for 10 min and extracted by liquid-liquid extraction with dichloromethane, then the nitrosamines were pre-concentrated using activated silica. Then, gas chromatography coupled with flame ionization detector was used for the separation and determination of the different nitrosamines contained in a real sample and gas chromatography with mass spectrometry detection was used as the confirmation technique. The newly invented autoclaving method allowed the determination of nitrosamine compounds at trace levels with limit of detection ranged from 0.077 to 0.18 ppb and quantitation limits were from 0.26 to 0.6 ppb for all nitrosamines, and found to be superior to the conventional ones, yielding approximately about 10%-20% increasing in the recovery compared with the mean recovery obtained when applying conventional methods.

Analysis of Nitrosamines in Organic Foods

Nitrosamines are classified by IARC as Group 2B carcinogens. Usually they might be present in organic foods as products of reaction between secondary amines and nitrosation system. The aim of the study was to test the concentration of nitrosamines in Bulgarian products. High performance liquid chromatography with UV detector was used for identification and quantitation. A standard solution of N-nitrosodiethanolamine was used as a reference substance and in the validation procedure of samples. The limit of detection of the method was determined to 14× 10^-9 g/mL. The results of the testing showed that analyzed organic foods produced in Bulgaria did not contain nitrosamines above the limit of detection of the method.

MECHANISM OF PROMOTING EFFECTS OF RIBOFLAVIN DEFICIENCY ON CARCINOGENESIS OF NITROSAMINES (EFFECTS ON RAT LIVER GLUTATHIONE CONTENT)

The effects of riboflavin deficiency and simultaneously nitrosodimethylamine (NDMA) given by gastric intubation on the hepatic glutathione (GSH) content were examined in rats. On different days of the experiment, hepatic GSH content of the riboflavin deficient rats decreased to 55-61% of the control rats. When NDMA was given 6 mg kg by gastric intubation to riboflavin deficient rats, hepatic GSH content decreased markedly to 39-43% of the control rats. After supplying riboflavin, hepatie GSH content of the deficient rats recovered to the level of the control rats. These results suggest that alterations of rat hepatic GSH content during riboflavin deficiency may imply as one of the promoting effects of riboflavin deficiency on the carcinogenesis of nitrosamines.

Carcinogenesis by Nitrosamines and Azoxyalkanes by Different Routes of Administration to Rats

The effects of administering similar doses of some simple alkylating agents to rats by different regimens have been compared.Two of the compounds were methylating agents,nitrosodimeth- ylamine and azoxymethane;two were ethylating agents,nitrosodiethylamine and azoxyethane and nitrosomethylethylamine was both a methylating and an ethylating agent.The treatments gave rise to tumors in almost all treated rats.The results indicate the importance ofpharmacoki- netics in determining which organs are the targets of the alkylating carcinogens.1989 Academic Press.Inc.

Application of Sargassum fusiforme(Harv.)Setch.Extract in Cigarettes

[Objectives]This study was conducted to investigate the application of Sargassum fusiforme(Harv.)Setch.in cigarettes.[Methods]Tobacco-specific nitrosamines in the smoke of cigarettes added with ethanol extract of S.fusiforme were determined,and the compounds related to the aroma of S.fusiforme were identified by flavor-smelling experiment.[Results]With the addition of the ethanol extract of S.fusiforme,the decrease in the total amount of four tobacco-specific nitrosamines in mainstream cigarette smoke reached 16.42%.The results of the flavor-smelling experiment showed that the aroma of S.fusiforme might be related to(R)-5,6,7,7A-tetrahydro-4,4,7A-trimethyl-2(4H)-benzofuranone,glycerol,ethyl palmitate,methyl palmitate,ethyl linoleate,methyl(Z,Z,Z)-9,12,15-octadecatrienoate,ethyl(Z,Z,Z)-9,12,15-octadecatrienoate,phytol,and tetradecanoic acid.[Conclusions]The ethanol extract of S.fusiforme has the potential function of reducing the content of tobacco-specific nitrosamines in smoke and improving the taste of cigarettes.

苦荞叶提取物清除亚硝酸钠及阻断亚硝胺合成作用研究

采用超声波辅助,响应面优化苦荞叶清除亚硝酸钠物质提取工艺,并以Vc和甘草提取物为对照,探究苦荞叶提取物对亚硝胺合成的阻断作用。结果表明,苦荞叶清除亚硝酸钠物质提取最佳工艺条件为:乙醇浓度44%,超声功率260 W,料液比21 mL/g,目数为80目,超声时间为30 min,提取温度50℃,此条件下苦荞叶提取物对亚硝酸钠的清除率为(83.97±0.12)%;在20~100 mg/L溶度范围内,Vc、苦荞叶提取物和甘草提取物对亚硝胺合成的阻断率线性关系良好,其对应的半阻断浓度IC50分别为61.26、80.09和90.20 mg/L。质量浓度继续增加时,苦荞叶提取物和甘草提取物对亚硝胺合成的阻断率随之下降,但同浓度情况下苦荞叶提取物对亚硝胺合成的阻断率高于甘草提取物,苦荞叶提取物对亚硝胺合成的阻断作用弱于Vc,但强于甘草提取物。

模拟胃液结合液质联用研究紫苏对内源性亚硝胺的阻断作用

为研究蔬菜对亚硝胺的阻断效果,在模拟胃液环境下结合液质联用,研究紫苏提取液对内源性亚硝胺的阻断作用。结果表明,在模拟胃液环境下加入紫苏液与亚硝酸钠,亚硝酸钠清除率可达10.4%。加入紫苏液、亚硝酸钠和二乙胺进行液相色谱质谱检测,空白样品和添加紫苏的样品中二乙基亚硝胺生成量分别为0.2、0.1 ng/mL,抑制率达到50%。添加紫苏的膳食中N-亚硝基二甲基(NDMA)、N-亚硝基二乙基(NDEA)、N-亚硝基二丁基(NDBA)、N-亚硝基吡咯烷(NPYR)含量显著降低。NDMA、NDEA在空白膳食中含量均为0.2 ng/g,紫苏试验组均未检出。NDBA空白对照为0.4 ng/g,紫苏试验组为0.2 ng/g,抑制率为50%;NPYR空白对照为8.6 ng/g,紫苏试验组为4.9 ng/g,抑制率为43%。因此,紫苏对内源性亚硝胺有明显的阻断作用。

DIANP合成及其在发射药中的应用研究进展

DIANP发射药作为一种新型发射药,具有能量高,爆温低,燃烧洁净性好,烟焰残渣少等特点。研究介绍了DIANP合成与材料特性,DIANP发射药配方及性能,DIANP发射药制备工艺、表面处理、装药应用和测试方法等方面的研究进展。指出了DIANP发射药在燃烧分解机理和相互作用机制等基础理论研究方面存在的不足;提出了应开展适用于DIANP发射药的温度系数控制技术研究、环境适应性研究和DIANP合成的绿色工艺研究;并认为DIANP发射药的改进需将理论研究和实际应用相结合,以此推进高能低烧蚀特征材料在行业内的发展。

水中N-亚硝胺的污染状况、来源及风险

N-亚硝胺类化合物是一类被国际癌症研究机构列为高毒性和强致癌性的有机污染物,是世界公认的除黄曲霉素、二噁英之外的第三大致癌物.N-亚硝胺水溶性强,广泛存在于环境水体(地表水、地下水)和饮用水当中.局部地区(特别是城市地区)环境水体及饮用水中的高浓度亚硝胺已对人体和生态健康构成较大威胁.除工业、农业和生活的一次排放外,前体物存在情况下的二次生成也是水体中亚硝胺持续存在的关键原因,某些情况下可能比一次来源更为重要.本文综述了环境水体及饮用水中N-亚硝胺的污染水平、组成和空间分布,给出水体中亚硝胺污染的重点化合物和重点区域;总结了导致亚硝胺二次生成的重要前体物,并归纳了N-亚硝胺在环境水体及饮用水系统中的一次来源及二次生成的机理过程,探索降低水体中亚硝胺污染的可行措施,为保障饮水安全和生态健康提供关键支撑.

基于网站服务器构建亚硝胺类基因毒性化合物数据库

目的:旨在构建一个全面的亚硝胺类杂质信息库,以助力研究人员方便地获取该类杂质的基本信息、毒性预测数据以及高分辨质谱数据。方法:收集可能存在于药物中的亚硝胺类化合物,利用PubChem等数据库对化合物的基本信息进行补充与完善。采用Derek和Sarah软件对化合物的基因毒性进行预测评估。同时,基于QTOF采集化合物的高分辨质谱数据。在完成数据的收集与整理后,运用Java编程语言搭建网站服务器,遵循MTV模式开发数据分析平台,实现数据信息的互联网共享。结果:成功构建了用户友好的亚硝胺类化合物信息查询平台。该平台能够清晰展示化合物的基本信息、基因毒性评估结果以及在标准条件下的质谱数据,方便研究人员快速了解化合物相关属性,并为后续的实验和研究提供参考。结论:所构建的亚硝胺类化合物分析平台不仅丰富了药物安全领域的数据资源,而且通过提供标准的分析结果,为药物的安全性和风险控制工作提供了数据支撑。

一步式QuEChERS-气相色谱-三重四极杆质谱法快速测定风干牦牛肉中15种N-亚硝胺

建立了一步式QuEChERS自动提取和净化技术结合气相色谱-串联质谱同时测定风干牦牛肉中15种N-亚硝胺的分析方法。样品水化后,经乙腈提取,加入4.0 g MgSO_(4)和1.0 g NaCl除水,经十八烷基硅烷(C_(18))和N-丙基乙二胺(PSA)填料净化,采用DB-HeavyWAX色谱柱(30 m×0.25 mm×0.25μm)分离15种N-亚硝胺,在多重反应监测(MRM)模式下进行测定,外标法定量。结果表明,15种N-亚硝胺分离性能良好,在0.1~200μg/L范围内线性关系良好,相关系数(r^(2))≥0.999 0;检出限(LOD)为0.05~0.20μg/kg,定量限(LOQ)为0.10~0.50μg/kg;在1倍、2倍和10倍LOQ 3个添加水平下的平均回收率分别为79.4%~102.1%、80.6%~109.5%、83.0%~110.6%,相对标准偏差(RSD)为0.8%~16.0%。应用建立的方法检测2种不同加工工艺的市售样品,其中7种N-亚硝胺类化合物(N-亚硝基二甲胺、N-亚硝基二异丁胺、N-亚硝基二正丁胺、N-亚硝基甲基苯胺、N-亚硝基乙基苯胺、N-亚硝基吡咯烷、N-亚硝基二苯胺)均有不同程度检出,平均含量为0.08~20.18μg/kg,且熟制风干牦牛肉中N-亚硝胺的检出率和平均含量均高于传统生制风干牦牛肉。该方法实现了前处理的自动化,相较于其他传统方法,操作简单,实验效率高,人为影响因素小,检测灵敏度高,适用于风干牦牛肉中15种N-亚硝胺的快速测定,为研究肉制品中N-亚硝胺的测定提供了方法支持。