金属食品罐内涂层中双酚类物质的迁移及检测研究进展

鉴于双酚A及其他双酚类物质作为食品罐内涂层材料有潜在的毒性与危害,中国、欧盟、美国等国家均已严格限制它们在金属食品罐内涂料中的使用。本文介绍双酚A、双酚A二缩水甘油醚、双酚F、双酚F二缩水甘油醚的结构、应用、危害、检测方法及迁移结果,并简述固相萃取在样品前处理中的应用。     

金属三片罐生产过程中双酚类物质迁移规律的研究

按照金属三片罐的实际生产工艺,跟踪研究了制罐内涂、外涂、彩印、上光、罐内补涂以及全喷工序过程的中间品,各自相应内壁涂膜中9种双酚物向3%(m/v)乙酸和10%(v/v)乙醇模拟液中的迁移情况。结果表明:双酚F二环氧甘油醚及其衍生物在两种模拟液中都未检出;而双酚A-(2,3-二羟丙基)甘油醚和双酚A-双(2,3-二羟丙基)甘油醚在两种模拟液中均有迁出;10%乙醇中还检测到双酚A二环氧甘油醚及其他衍生物。在10%乙醇体系中,迁出的双酚物在全喷后增加显著。3%乙酸体系中,迁出的双酚物在补涂和全喷后增加明显。结果表明补涂与全喷这两个工艺对三片罐的食品安全性造成潜在影响,可在加工过程中作为关键控制点进行控制。     

食品金属罐中14种双酚类物质迁移量的同时测定及杀菌过程对其迁移的影响

为了调查国内食品金属罐中双酚类物质的迁移水平,本文建立了同时测定食品金属罐中14种双酚类物质迁移量的高效液相色谱法,并研究了杀菌过程对双酚类物质迁移的影响及食品金属罐中双酚类物质的迁移规律。结果表明,14种双酚类物质在4种食品模拟物（4%体积乙酸、10%体积乙醇、50%体积乙醇和异辛烷）中线性关系良好,加标回收率为83.67%～107.05%,精密度范围为2.32%～7.67%,该方法的精密度和准确度较好,可用于食品金属罐中14种双酚类物质迁移量的检测分析。对市场上9种罐头和饮料用金属罐中的14种双酚类物质的迁移量进行了同时测定,发现有7种双酚类物质检出,无双酚A及其类似物的检出。基于欧盟限量标准要求,发现午餐肉罐头中的双酚A-（2-3-二羟基丙基）缩水甘油醚（BADGE·H₂O）、牡蛎葛根饮料中的双酚A-二（2-3-二羟基丙基）醚（BADGE·2H₂O）和黑莓罐中的双酚F-（2-3-二羟基丙基醚）（BFDGE·2H₂O）存在一定的迁移风险。此外,杀菌过程影响食品金属罐中双酚类物质的迁移种类和迁移量。迁移试验结束后,经过杀菌的金属罐中仅检测出BADGE·2H₂O,而未经杀菌的金属罐中同时检测到BADGE·2H₂O和双酚A-（3-氯-2羟丙基）（2-3-二羟基丙基）缩水甘油醚（BADGE·H₂O·HCl）。另外,食品模拟物的类型影响着双酚类物质的迁移量和迁移种类,且随着迁移温度的升高和迁移时间的增加,双酚类物质的迁移量逐渐升高直至平衡。     

国内市场上罐装啤酒中双酚类物质迁出量调查

建立并利用固相萃取-高效液相-荧光法对国内市场上31种罐装啤酒中双酚物的含量进行了调查。方法的检测限为0.02～0.09μg/L,在0.02～50.00μg/L的检测范围内相关系数大于0.99,加标回收率和标准差分别为80.67%～101%和1.01%～4.84%。样品中除双酚E(BPE)没有检出外,双酚A(BPA)、双酚A二缩水甘油醚(BADGE)、双酚F二缩水甘油醚(BFDGE)、双酚B(BPB)和双酚F(BPF)的平均检出浓度分别为2.99、0.41、0.18、0.12、0.12μg/L,检出率分别为100%、83.9%、6.4%、12.9%和6.4%。BPA和BADGE是罐装啤酒中存在的最主要的双酚物,其平均每日摄入量分别估计为0.0160、0.0022μg/kg.bw.d,低于欧盟关于BPA50μg/kg.bw.d的规定。样品中BPA没有超过欧盟规定的0.6mg/L,BADGE的迁移也未高于9mg/L,但是BFDGE的检出是不允许的。目前尚无关于BPF、BPE和BPB的迁移限值。     

食品模拟物中双酚A迁移量的间接竞争免疫PCR检测方法

通过抗原-抗体的特异性反应,结合荧光PCR,建立了食品接触材料中双酚A在食品模拟物中迁移量的快速、灵敏的间接竞争免疫PCR检测方法。通过优化包被抗原浓度、探针DNA的量、抗体质量浓度等一系列条件,双酚A的线性质量分数范围在0.01~10 pg/g之间,检测低限为3.0 fg/g,4种食品模拟物中双酚A的添加回收率在82.8%~115.2%,该方法特异性好、灵敏度高,适用于食品接触材料中双酚A及其他小分子化学物的痕量迁移检测,为其迁移行为及迁移模型的建立提供了技术支撑。     

肉类罐头食品中双酚A二缩水甘油醚及其衍生物的迁移

本实验应用高效液相色谱-电喷雾串联质谱法分析检测肉类罐头食品中双酚A二缩水甘油醚(BADGE)及其衍生物的残留,重点研究了肉类罐头中的内容物、存储时间(6个月、9个月、12个月)以及存储温度(4、20、100℃)对双酚A二缩水甘油醚(BADGE)及其衍生物的迁移量的影响,单因素方差分析(One-way ANOVA)检验实验结果表明,不同内容物罐头中从内壁涂层迁移至样品的BADGE及其衍生物存在显著性差异(p<0.05),Student-Newman-Keuls法检验表明存储12个月后目标化合物的迁移量与6个月、9个月存在显著性差异,存储温度为4、20℃时化合物迁移量没有显著差异,但是罐头加热到100℃后目标化合物迁移量是最大的。     

食品包装材料中双酚A迁移量的测定

建立采用高效液相色谱法对塑料食品包装材料中双酚A向食品模拟物迁移量的检测方法。使用4种食品模拟物：水、质量分数为4%乙酸溶液、体积分数30%的乙醇溶液和脂肪类模拟物(正己烷、异辛烷和橄榄油)。结果表明,双酚A在与食品塑料包装接触过程中,无论在何种情况下都会向食品模拟物中迁移,尤其向醇类模拟物中迁移最严重;在温度超过60℃时,双酚A向食品模拟物中的迁移率骤增;在微波加热条件,高火700W功率时双酚A向食品模拟物中的迁移速率最快。该方法检测限为0.3ng/mL,线性范围为0.5～100ng/mL,线性相关系数为0.9997,回收率在92.0%～102.4%之间,相对标准偏差≤2.84%(n=5)。     

石墨烯固相萃取-液相色谱-质谱联用法测定食品接触材料中9种双酚-二环氧甘油醚的迁移量

基于石墨烯固相萃取,以实际样品为研究对象,建立了食品接触材料中内分泌干扰物双酚A-二缩水甘油醚(BADGE)及其衍生物BADGE·H2O、BADGE·HCl、BADGE·2H2O、BADGE·2HCl、BADGE·H2O·HCl和双酚F-二缩水甘油醚(BFDGE)及其衍生物BFDGE·2HCl、BFDGE·2H2O等9种双酚-二环氧甘油醚迁移量的高效液相色谱-串联质谱(HPLC-MS/MS)检测方法。样品以叔丁基甲醚为提取溶剂,超声提取,提取液经石墨烯固相萃取柱富集、净化。质量分数0.1%乙酸的5 mmol/L醋酸铵溶液和甲醇为流动相,经COSMOSIL C18分析柱分离后,采用电喷雾串联四级杆质谱检测。9种双酚-二环氧甘油醚中2种在0.5~50 ng/m L范围内线性关系良好;2种在5.0~100.0 ng/m L范围内线性关系良好;其他5种在2.0~100.0 ng/m L范围内线性关系良好(r20.992 2)。对牛肉、橘片两种罐头食品进行3个水平的添加回收实验,9种目标化合物的回收率分别为:60.0%~97.6%(牛肉)和65.7%~111.3%(橘片),相对标准偏差小于20%。方法的检测低限为0.1~1.0μg/kg。该方法操作简单,灵敏度高,成功应用于食品接触材料中双酚-二环氧甘油醚迁移量的快速检测。     

罐头涂料铁罐双酚A二缩水甘油醚及其衍生物迁移规律研究

研究了罐头涂料铁罐双酚A二缩水甘油醚及其衍生物在特定食品模拟物中的迁移规律。采用50%(v/v)乙醇溶液为食品模拟物,应用高效液相色谱-串联质谱法进行双酚A二缩水甘油醚及其衍生物的定性定量分析,并应用基于Fick扩散第二定律的D.Chung模型和基于概率与数理统计方法的Weibull模型进行了预测。结果表明:贮存条件对迁移总量变化影响显著,温度越高越快达到平衡,迁移平衡值为1.28 mg/kg;D.Chung模型中扩散系数D与贮存温度符合阿伦尼乌斯动力学模型,该模型预测值与实验值的拟合优度(R~2)小于0.23,而Weibull模型拟合优度(R~2)能达到0.90以上。因此,Weibull模型比D.Chung模型更接近实际迁移结果,为评估涂料铁罐的安全性提供参考。     

LC-MS/MS分析罐装食品和饮料中的双酚A-缩水甘油醚(BADGE)、双酚F-缩水甘油醚(BFDGE)及其衍生物

双酚A—缩水甘油醚(BADGE)和双酚F—缩水甘油醚(BFDGE)经常被用于罐装食品和饮料中,但它们及它们的衍生物存在着潜在的致突变性、遗传毒性和抗雄激素作用,欧盟已对其总量作了限制。本文的目的就是开发一种能同时定量测定罐装食品和饮料中BADGE、BFDGE及其衍生物的快速灵敏的LC—MS/MS方法。     

Food and beverage can coatings: A review on chemical analysis,migration, and risk assessment

The internal surface of food and beverage cans is generally covered with polymeric coatings to preserve food and protect metal substrate from corrosion. Coating materials are complex formulations that contain different starting substances (e.g., monomers, prepolymers, additives, etc.) and in addition during the manufacture of the material several compounds can be formed (e.g., reaction products, degradation products, etc.). These substances have the potential to migrate into the food. Many of them have not been identified and only some have been toxicologically evaluated. This article aims to provide a comprehensive review on the analytical methods used for the identification of potential migrants in can coatings. The migration and exposure to chemicals migrating from can coatings are also reviewed and discussed so far, which is essential for risk assessment. Moreover, a brief section on the current status of the legislation on varnishes and coatings for food contact in Europe is also presented. Liquid chromatography coupled to diode array and fluorescence detectors and particularly to mass spectrometry and gas chromatography–tandem mass spectrometry seem to be the techniques of choice for the identification of potential migrants in can coatings. Some studies have reported migration levels of BPA (bisphenol A) and BADGE (bisphenol A diglycidyl ether) and derivatives exceeding the specific migration limits set in the European legislation. On the whole, low dietary exposure to migrants from can coatings has been reported. However, it is interesting to highlight that in these studies the combined exposure to multiple chemicals has not been considered.     

Improved sample extraction and clean-up for the GC-MS determination of BADGE and BFDGE in vegetable oil.

A straightforward method was established for the determination of migration contaminants in olive oil with a special focus on the two can-coating migration compounds bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE). The preferred sample preparation was a single liquid-liquid extraction of compounds from the oil into 20% (v/v) methanol in acetonitrile, followed by clean-up with solid-phase extraction on aminopropyl bonded to silica. This purification procedure selectively removed all free fatty acids from the extracts without removing phenolic compounds of interest. The solid-phase extraction columns were used many times by implementing a procedure of washing out the strongly retained fatty acids with 2% acetic acid in methanol. Gas chromatography coupled with full scan (m/z 33-700) electron ionization mass spectrometry was used for the determination of several model compounds in olive oil samples. BADGE and BFDGE could be determined in the 0.05-2 mg kg(-1) range in oil samples with a relative SD of <6% (six replicates). The method was used in an enforcement campaign for the Norwegian Food Control Authority to analyse vegetable oil samples from canned fish-in-oil.     

快餐包装纸中矿物油向固体食品模拟物的迁移

研究从市场上收集到的24 种快餐包装纸中的矿物油向固体食品模拟物Tenax的迁移规律。在多个不同的迁移条件（40 ℃/0.5、1、2、3 h,40 ℃/10 d和70 ℃/2 h）,探究矿物油的迁移行为及其影响因素,以评价其安全性。选择正己烷-乙醇（1∶1,V/V）混合溶液对Tenax进行过夜萃取,采用质量分数0.3%硝酸银固相萃取柱对饱和烃矿物油（mineral oil saturated hydrocarbons,MOSH）和芳香烃矿物油（mineral oil aromatic hydrocarbons,MOAH）进行分离纯化,最后用气相色谱-氢火焰离子化检测法和气相色谱-质谱法分别进行定量和定性分析。结果表明：随着温度的升高,多种快餐包装纸中矿物油向Tenax的迁移量也随之增加。涂蜡纸中MOSH迁移量均有所检出,其数值为110.49～615.40 mg/kg,而MOAH部分均未检出,这可能是因为涂蜡纸表面涂覆的石蜡层属于MOSH类,导致其MOSH部分的迁移量较高。网购餐盘纸和常规餐盘纸中MOSH迁移量约为其特定限量值（0.6 mg/kg）的10～400 倍,MOAH的迁移量约为其特定限量值（0.5 mg/kg）的10～70 倍,而使用优质胶印油墨的原生纤维餐盘纸均未超过其限量值。最后,通过对印有胶印油墨的原生纤维餐盘纸中矿物油进行溯源分析,发现经过迁移的矿物油一部分可能来源于其所用油墨,其他来源可能来自于回收纤维、黏合剂、添加剂和加工助剂等。     

Migration from can coatings: Part 3. Synthesis, identification and quantification of migrating epoxy-based substances below 1000 Da

Bisphenol A-derived glycidyl ethers as well as its reaction products with other lacquer components can migrate into the packed food from epoxy-based can coatings. A sensitive and selective method is presented using high-performance liquid chromatography coupled with ultraviolet light, fluorescence and electrospray ionization-mass selective detection for the identification and quantification of all migrants with a bisphenol A backbone and a molecular weight below 1000 Da, an estimated boundary for the absorption in the gastrointestinal tract. The identification of migrants was confirmed by microreactions of technical bisphenol A diglycidyl ether with solvents and phenols, which provided the fragmentation pattern of the mass selective detection and relative retentions of 42 different bisphenol A-related substances. It was shown by calibration of different isolated and synthesized bisphenol A derivatives that the fluorescence response relies on the amount of bisphenol A moiety in the respective molecule. Therefore, all migrating bisphenol A-related substances below 1000 Da were determined as bisphenol A diglycidyl ether equivalents using a calibration (fluorescence detection) of the commercially available bisphenol A diglycidyl ether monomer. The limit of quantification was set at 5 μg bisphenol A diglycidyl ether equivalents kg^-1 (or 0.8 μg dm^-2). This method was validated for epoxy coatings (0.1 μg dm^-2 limit of detection and 24 μg bisphenol A-related substances below 1000 Da dm^-2 standard deviation, corresponding to 4.4% relative standard deviation). The quantification could be extended by combining the fluorescence response and structural information gained from the mass spectra, which provides more accurate results for each migrant. The calculation is based on the calibration of the bisphenol A chromophore content of the molecule. According to this method, the amount of migrating bisphenol A-related substances below 1000 Da in the acetonitrile extract (assuming a worst case) varied from about 0.4 to 0.7 mg dm^-2 in the examined coatings. The determined amounts comply with about 50% of the total migrate below 1000 Da.     

三片食品罐外壁涂膜中有害物质向内涂的迁移

对制版印刷后的食品用三片金属罐外壁涂料中三聚氰胺、苯、甲苯3种有害物质在堆码过程中从外壁涂膜向内涂膜的迁移扩散进行研究。采用高效液相色谱法检测3种常用金属罐(番茄酱罐、八宝粥罐、凉茶罐)内壁和外壁涂膜中三聚氰胺、苯和甲苯3种残留物在去离子水、3g/100mL乙酸、体积分数10%乙醇3种水性食品模拟液中的初始迁移量。并对制罐铁片在恒温(30℃)、恒湿(60±2)% RH、恒压34kPa条件下堆码2个月后内壁涂膜中3种残留物的向食品模拟液中的迁移量与堆码前进行比较分析,即外壁涂膜中这3种残留物向内壁的迁移扩散情况。结果表明,3种有害物质均有从外壁向内壁迁移的现象,迁移量达到0.04～1.60μg/mL,对内壁涂膜的安全性造成极大影响,严重影响到罐装食品的安全性。     

A LC-MS/MS method for the determination of BADGE-related and BFDGE-related compounds in canned fish food samples based on the formation of [M+NH(4)](+) aducts

A new and simple liquid chromatography tandem mass-spectrometry method for the determination of different bisphenol A (BPA) derivatives such as bisphenol A diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their reaction products with water and hydrochloric acid in different fish food products was developed. The extraction procedure and the chromatographic conditions were optimised for complex food matrices such as fish products. Food samples were homogenised and extracted with a 1:1 solution of acetonitrile-hexane, the solvent was eliminated in a N(2) stream and the extract was reconstituted with 0.5mL of a 0.01M solution of ammonium formate. The sample solution obtained was directly measured by LC-MS/MS without any further purification under the developed conditions. The use of a mobile phase composed by ammonium formate-methanol in a binary gradient mode produced [M+NH(4)](+) aducts for the different BADGEs and BFDGEs. These aduct's fragmentations were employed for the LC-MS/MS quantification of BPA derivatives in canned fish samples. The results of the validation were appropriate: the method was linear for BADGE and its hydrolysed derivatives up to 1000μgkg(-1), for the remaining compounds linearity achieved up to 100μgkg(-1). Quantification limits were in the range 2-10μgkg(-1). RSD (intra and inter-day) was 6-12% and the recovery was comprised between 89% and 109%. Under the optimised conditions, the chromatographic separation was performed in 8min per sample. The method was applied to the determination of BADGE, BFDGE and their reaction products in different samples of canned fish from Spanish origin. Migration results obtained were in compliance with the EU regulations.     

Stability of bisphenol A diglycidyl ether and bisphenol F diglycidyl ether in water-based food simulants

Varnishes used for the inner coatings of food cans are often based on epoxy resins or vinylic organosols. The epoxy resins can be produced from bisphenol A (BPA) and bisphenol F (BPF), and these also contain bisphenol A diglycidyl ether (BADGE) or bisphenol F diglycidyl ether (BFDGE) as stabilising components. These compounds may break down during storage and also by the influence of food simulants. The stability of BADGE and BFDGE were studied using reverse-phase gradient high-performance liquid chromatography (RP-HPLC) with ultraviolet detection (UV). Three experimental conditions for spiked simulants were compared: (1) the storage at 25 °C (C1), (2) the storage at 40 °C (C2) and (3) the storage at 25 °C after 15 min heating at 120 °C (C3). Distilled water, 3% acetic acid and 10% ethanol were used as food simulants. It was observed that BADGE is more stabile than BFDGE. The loss of BADGE and BFDGE were minimal in 10% ethanol (39 and 46% at 25 °C, 60 and 69% at 40 °C, respectively) and highest in 3% acetic acid (60 and 63% at 25 °C, 76 and 82% at 40 °C, respectively). At experiment (C3), the hardest conditions, significant degradation was not shown in comparison with conditions (C1) and (C2), contrariwise BADGE and BFDGE in 10% ethanol were minimal degradated at conditions (C3) from all these experiments (loss of 5 and 8%, respectively).     

食品接触材料AS/ABS中14种苯类化合物迁移量和风险评估模型的建立

为建立顶空-气相色谱质谱联用仪测定AS/ABS产品在水、4%乙酸、10%乙醇、20%乙醇、50%乙醇和95%乙醇模拟物中,14种苯类化合物特定迁移量(乙醇模拟物为13种苯类化合物迁移量)的定性和定量检测方法,通过优化分流比、载气流速、程序升温速度、平衡温度和时间等色谱条件,采用DB-WAXMS(30 m×0.25μm×0.25 mm)色谱柱,实现了苯、二甲苯、三甲苯以及丙苯等苯类的有效分离,14种苯类化合物迁移量的定量限(10S/N)为0.6~2.1μg/kg,0.20 mg/kg的混合标准溶液回收率为80.2%~118.3%,RSD值为3.6%~9.7%,线性相关系数均大于0.9950,实现了对AS/ABS中14种苯类化合物特定迁移量的快速定性和定量分析。试验还考察不同温度和时间条件下食品接触产品AS/ABS中苯类化合物的迁移量规律,结果显示,温度越高,苯类化合物特定迁移量越大。建立了不同温度下苯类化合物迁移模型,确定了扩散系数DP值、特征参数Ap值、活化能Ea等动力学参数,建立了食品接触材料AS/ABS中苯类化合物风险评估数学模型,为AS/ABS中苯类化合物风险管控提供了依据。