Determination of benzidine in the food colours tartrazine and sunset yellow FCF, by reduction and derivatization followed by high-performance liquid chromatography.

Free and bound benzidine, a non-sulphonated aromatic amine (NSAA), were determined in the food colours tartrazine and sunset yellow FCF. Bound amines were released by reducing with sodium dithionite, then total NSAAs were extracted into chloroform, transferred to aqueous acid solution and diazotized with sodium nitrite before coupling with 2-naphthol-3,6-disulphonic acid, disodium salt (R-salt). Coloured benzidine and aniline derivatives (BZDRS and ANRS) were analysed using reversed-phase ion pair high-performance liquid chromatography (HPLC) and an absorbance detector set at 548 nm. Levels of total benzidine were similar to those reported in studies conducted in the USA, and ranged from < 5 to 270 ng/g. Total aniline was also determined (0.2-188 micrograms/g). Recoveries of benzidine with this method were found to be lower than expected (average ca 46%), but were reproducible. Detection limits were 15-20 ng BZDRS/g (3-4 ng benzidine/g). Mass spectrometry (LC-MS) was evaluated for identifying and determining purity of the standards, but difficulties were encountered when the methodology was applied to commercial samples.     

Determination of total non-sulphonated aromatic amines in soft drinks and hard candies by reduction and derivatization followed by high-performance liquid chromatography.

Utilizing elements of methodology developed previously for food colours, total free and bound non-sulphonated aromatic amines (NSAA) were determined in commercial samples of soft drink beverages and hard candies. Bound amines in the samples were reduced using sodium dithionite, then total NSAA were extracted into chlorofom, transferred to aqueous acid solution and diazotized with sodium nitrite before coupling with 2-naphthol-3,6-disulphonic acid, disodium salt (R-salt). The coloured derivatives were analysed using reversed-phase ion pair high-performance liquid chromatography (HPLC) and an absorbance detector set at 512 nm. Solid phase extraction cartridges were utilized for extraction and clean-up of the food colours present in the sample, and the concentration of each dye was determined quantitatively using HPLC and absorbance detector wavelengths of 426, 516 or 625 nm. Levels of total NSAA were compatible with those observed previously in food colours. Commercial soft drinks were found to contain (expressed in terms of total free plus bound NSAA in the beverage) 0.19-12.6 ng/ml of aniline, 0.83-8.25 ng/ml 1-naphthylamine and 0.62-1.12 ng/ml 2-naphthylamine. Levels of 0.66-9.15 ng/g of aniline and 2.48-10.6 ng/g 1-naphthylamine were found in commercial samples of hard candies. Bound NSAA in hard candies appeared to survive the manufacturing process. Recoveries averaged 96.9% for tartrazine and 89.6-97.2% for the bound amines when hard candies were prepared in the laboratory.     

常用色素的护色剂选择研究

在分别研究三聚磷酸钠、焦磷酸钠、六偏磷酸钠、柠檬酸钠、EDTA-2Na、植酸、异抗坏血酸钠、乳酸钙等护色剂对胭脂红、苋菜红、诱惑红、柠檬黄、日落黄、亮蓝、β-胡萝卜紊等单一护色作用的基础上，选择护色效果较好的护色剂．通过正交设计，优选复配护色剂的配方。结果表明，这些护色剂的护色作用明显，但EDTA-2Na对日落黄、异抗坏血酸钠对大部分色素有负作用；各色素较好的复配护色剂组成，胭脂红为植酸2／万、三聚磷酸钠2／万、EDTA-2Na1／万、六偏磷酸钠1／万，苋菜红为EDTA-2Na2／万、焦磷酸钠1／万、三聚磷酸钠2／万及植酸2／万，诱惑红为三聚磷酸钠2／万、六偏磷酸酸2／万、EDTA-2Na2／万及焦磷酸钠2／万，日落黄为植酸1／万、焦磷酸钠2／万、柠檬酸钠2／万及六偏磷酸酸1／万，β-胡萝卜素为异抗坏血酸钠1／万、EDTA-2Na2，万、植酸1／万及焦磷酸钠1／万。     

2012~2015年贵州省餐饮食品中色素检测结果分析

目的 对贵州省餐饮服务食品中柠檬黄、诱惑红、日落黄、胭脂红和苏丹红5种色素进行检测与分析, 掌握我省餐饮食品安全状况。方法 根据GB/T 5009.35-2003《食品中合成着色剂的测定》, SNT 1743-2006《食品中诱惑红、酸性红、亮蓝、日落黄的含量检测 高效液相色谱法》和GB/T 19681-2005《食品中苏丹红染料的检测方法 高效液相色谱法》, 对2012~2015年贵州省餐饮服务食品中5种色素进行检测, 按GB/2760-2011《食品添加剂使用卫生标准》和《食品中可能违法添加非食用物质名单》判断是否合格。结果 3054批食品中, 柠檬黄、诱惑红、日落黄、胭脂红和苏丹红的合格率分别为95.6%、99.1%、98.5%、99.3%和100%。结论 贵州省4年内餐饮服务食品中色素的总体安全状况较好。     

超高效液相色谱法同时测定包衣糖果中5种铝色淀

目的 建立包衣糖果中柠檬黄、日落黄、胭脂红、诱惑红和亮蓝5种铝色淀的超高效液相色谱测定方法.方法 用NaOH溶液浸泡提取包衣糖果中的色淀,以乙腈-乙酸铵(10 mmol/L)为流动相,梯度洗脱,多波长检测定量.结果 5种目标化合物在0.25～50 mg/L范围内呈良好的线性,相关系数R2 ＞0.999.该方法中5种目标化合物的定量限(LOQ)均为1.0 mg/kg.在1、5、25 mg/kg三个添加浓度水平下,5种目标化合物的加标回收率为79.5％ ～116.4％,RSD为0.92％ ～6.48％.结论 该方法简单、快速、准确,适于包衣糖果中5种铝色淀的同时检测.     

调味品中合成着色剂和糖精钠的测定

目的 建立了一种同时测定调味品中合成着色剂（诱惑红、胭脂红、日落黄）和糖精钠的高效液相色谱（HPLC）分析方法.方法 采用C18色谱柱,以甲醇和0.02 mol/L乙酸铵溶液为流动相,梯度洗脱,光电二极管阵列（PDA）检测器检测,采用外标法定量分析.结果 4种成分的线性范围为0.5～20.0 μg/mL （r＞0.999）,回收率为94.3％～101.6％,RSD为0.42％～2.44％（n=6）.结论 此法准确、灵敏,前处理简单,适用于调味品中诱惑红、胭脂红、日落黄和糖精钠的定性、定量分析.     

Stability of sunset yellow FCF and tartrazine in liquid model food systems

Effect of various preservatives, sugar, ascorbic acid and light on the stability of sunset yellow and tartrazine was studied. While sulphur dioxide enhanced the rate of degradation of sunset yellow, it stabilized tartrazine in aqueous model systems. At 30 and 45% sugar levels, the rate of degradation of both sunset yellow and tartrazine was practically same in the presence of benzoic acid and sorbic acid. At 15% sugar level, the rate of degradation of tartrazine was slightly higher in the presence of sorbic acid. Incorporation of ascorbic acid enhanced the degradation of both sunset yellow and tartrazine and its effect was considerably higher in the presence of light. Increase in sugar concentration enhanced the stability of both the dyes, but light enhanced their degradation.     

高胶质食品中合成色素检测方法的改进

采用用聚酰胺吸附法、碱液直接提取法、碱性醇液提取法、饱和硼砂沸腾水浴提取法、海砂研磨法5种提取方法分别提取凤爪中的柠檬黄、苋菜红、胭脂红、日落黄、诱惑红和亮蓝6种色素,并用超高效液相色谱法测定。结果表明:5种提取方法中,海砂研磨法对凤爪中6种色素的提取效率最高;用选定的方法对超市中抽检的25个凤爪样品进行检测,发现样品中主要含有柠檬黄、日落黄和诱惑红,含少量胭脂红,不含苋菜红和亮蓝。     

高效液相色谱法测定明胶胶囊中11种合成色素的方法研究

建立一种用高效液相色谱(high performance liquid chromatography,HPLC)同时测定明胶胶囊中的11种合成色素的分析方法。样品经木瓜蛋白酶60℃水解30 min后,以CAPCELL PAK C₁₈柱(4.6 mm×250 mm,5 μm)为分析柱,甲醇-乙腈(1:1)和15 mmol/L乙酸铵为流动相,梯度洗脱进行分离,以二极管阵列检测器检测,检测波长450(柠檬黄、喹啉黄)、520(新红、苋菜红、胭脂红、日落黄、诱惑红、偶氮玉红、赤藓红)和620 nm(靛蓝、亮蓝)。结果表明:11种合成色素在0~100 μg/mL浓度范围内线性关系良好,相关系数均大于0.999,方法检出限为0.3~2.5 mg/kg,回收率在79.1%~119.3%之间。该方法操作简便,灵敏度高,分离效果好,能实现11种色素的同步测定,适用于明胶胶囊中合成色素的检测分析。     

Determination of Amaranth in Beverage by Indirect Competitive Enzyme-linked Immunosorbent Assay (ELISA) Based on Anti-amaranth Monoclonal Antibody

Amaranth (E 123) is a member of azo dyes, and it is allowed to use in various foods. The acceptable maximum addition of amaranth is strictly fixed because of its potential risk to physical health. The objective of this study was to prepare a specific anti-amaranth monoclonal antibody and develop an indirect competitive ELISA for amaranth quantification analysis. The immunogen and the coating antigen were designed by introducing a carboxyl group into amaranth for the conjugation with carrier proteins. Based on the immunogen, the monoclonal antibody exhibits satisfactory performances and the proposed ELISA shows an IC₅₀ of 20.33 ng mL^?1. The limit of detection is as low as 3.35 ng mL^?1, and the linear standard curve of the method ranges from 3.0 to 243.0 ng mL^?1. Additionally, the antibody reflects minimal cross-reactivity (<1 %) with six related food dyes (erythrosine, ponceau 4R, allura red, tartrazine, sunset yellow FCF, and brilliant blue). The recoveries of amaranth spiked beverage samples are in the range of 85.8–100.7 % with low coefficient of variation values (<11.5 %). The data shows that the developed ELISA provides a simple, sensitive, specific, and accurate alternative for amaranth determination and monitoring. Furthermore, it is the first time that icELISA of amaranth is developed based on monoclonal antibodies.     

基于酸度控制的吸收光谱法测定饮料中共存的柠檬黄和日落黄

建立快速、准确测定饮料中共存着色剂柠檬黄与日落黄的吸收光谱检测方法。在一定酸度的Tris-HCl介质中，乙基紫与柠檬黄（pH 5.69）和日落黄（pH 8.68）反应生成离子缔合物，在波长506 nm处产生能定量分析共存色素中柠檬黄的特征吸收峰，在波长646 nm处产生能定量分析共存色素中日落黄的特征吸收峰，它们的表观摩尔吸光系数分别为1.37×105 L/（mol·cm）（柠檬黄体系）和7.68×104 L/（mol·cm）（日落黄体系）。柠檬黄在0.04～9.6 mg/L范围内与吸光度绝对值呈线性关系，检出限为0.024 mg/L，定量限为1.29 mg/kg；日落黄在0.04～6.3 mg/L范围内与吸光度呈线性关系，检出限为0.031 mg/L，定量限为1.68 mg/kg。该法用于实际样品分析，加标回收率为97.0%～104%（柠檬黄体系）和97.3%～103%（日落黄体系），相对标准偏差（n=5）为1.5%～2.6%（柠檬黄体系）和1.2%～2.6%（日落黄体系）。该法简便、快速，用于饮料中共存柠檬黄和日落黄的测定，结果满意。     

甲基绿吸收光谱法同时测定饮料中的酒石黄和日落黄

研究建立了一种简便、快速、能同时测定饮料中共存着色剂——酒石黄和日落黄的吸收光谱新方法。在pH 9.43 Tris-HCl溶液中,甲基绿与酒石黄和日落黄发生显色反应,生成离子缔合物,在400~750 nm范围内,酒石黄体系在574 nm波长处产生1个能进行定量分析的特征负吸收峰,共存色素日落黄不干扰测定,酒石黄的质量浓度在0.04~16.0 mg/L范围内与缔合物的吸光度绝对值(A)有很好的线性关系,表观摩尔吸光系数为4.90×10⁴ L/(mol·cm),检出限为0.033 mg/L,饮料的定量限为1.80 mg/kg;日落黄体系在488 nm波长处产生1个能用于定量分析的特征正吸收峰,共存色素酒石黄不干扰测定,日落黄的质量浓度在0.04~13.6 mg/L范围内与缔合物的吸光度有很好的线性关系,表观摩尔吸光系数为4.19×10⁴ L/(mol·cm),检出限为0.028 mg/L,饮料的定量限为1.51 mg/kg。该方法简便、快速,通过控制溶液酸度便可在不经分离的情况下同时测定同一饮料样品中共存色素——酒石黄和日落黄的含量。     

高效液相色谱法测定饮料中12种水溶性合成着色剂

目的建立高效液相色谱法同时测定饮料中柠檬黄、喹啉黄、苋菜红、胭脂红、日落黄、诱惑红、酸性红、赤藓红、靛蓝、酸性绿S、亮蓝、专利蓝V 12种水溶性合成着色剂的方法。方法饮料样品经离心、过滤后直接进样,以甲醇-乙酸铵(0.02 mol/L)为流动相进行梯度洗脱,采用二极管阵列检测器多波长分析,外标法定量。结果 12种合成着色剂在0.5～100mg/L范围内线性关系良好,相关系数均大于0.999;检出限为0.05～0.3 mg/kg;平均回收率为91.0%～101.3%,相对标准偏差值为0.2%～2.3%。结论该方法简便、快速、准确,前处理简单,适合于基质成分较为简单的液体样品中多种着色剂的同时测定。     

双波长比值光谱法测定饮料中的色素

应用双波长比值光谱法测定了果味汽水、水果泡泡中胭脂红、日落黄、酒石黄的含量。依据该3 种色素的比值光谱特征,以日落黄为干扰组分,分别选择431、456nm 和509、525nm 为测定酒石黄和胭脂红的波长;以胭脂红为干扰组分时,选择514、535nm 为测定日落黄的波长。结果显示,酒石黄在0.1～60mg/L、胭脂红在0.2～80mg/L、日落黄在0.8～90mg/L 的质量浓度范围内线性关系良好,平均回收率分别是100.5%、101.1%和102.4%。本方法操作简便、易于推广,可应用于实际样品的检测。     

离心辅助提取果冻中合成着色剂

建建立了一种快速分离、提取果冻中人工合成着色剂的方法。将聚酰胺粉与果冻加热混匀、离心,聚酰胺粉能吸附合成着色剂并沉降于底部,与果胶能很好的分离。采用国标方法进行洗脱、解析,可有效的提取出果冻中的合成着色剂。结果表明,柠檬黄与日落黄在2.5μg/mL～50μg/mL范围内有良好的线性关系,标准曲线相关系数均达到0.9999以上;加标回收率分别为96.62%和94.10%;RSD分别为2.21%和2.26%。     

HPLC法检测蜜饯中20种合成着色剂含量

建立蜜饯中柠檬黄、新红、苋菜红、靛蓝、丽春红S、胭脂红、喹啉黄、日落黄、诱惑红、亮蓝、酸性红44、酸性红、食品红1、橙黄1、酸性红50、专利蓝V、赤藓红、酸性橙2、酸性橙8、亮蓝G共20种合成着色剂的高效液相色谱测定方法。试样用甲醇/氨水溶液提取,混合型弱阴离子反相固相萃取柱净化,甲醇/20 mmol/L乙酸铵为流动相梯度洗脱,二极管阵列检测器多波长检测,外标法定量。结果表明,该方法在0.1~20 μg/mL的线性范围内相关系数均大于0.999,加标回收率范围为84.0%~104.5%,相对标准偏差（RSD）为1.3%~5.3%。该方法操作便捷、稳定性好、回收率高,适用于蜜饯中20种合成着色剂的同时分析检测。该方法可以扩展应用于其他同结构性质的色素分析,对其他食品类别中色素的检测也具有参考意义。     

Estimates of dietary exposure of children to artificial food colours in Kuwait

To assess the intake of artificial food colour additives by 5-14-year-old children in the State of Kuwait, a 24-h dietary recall was conducted twice on 3141 male and female Kuwaiti and non-Kuwaiti children from 58 schools. The determination of colour additives in 344 foods items consumed was performed using high-performance liquid chromatography with diode array detector. A comparison with the Food and Agriculture Organization and World Health Organization acceptable daily intakes (ADIs) was undertaken to evaluate the potential risk associated with the consumption of artificial colour additives by children in Kuwait. The results indicated that out of nine permitted colours, four exceeded their ADIs by factors of 2-8: tartrazine, sunset yellow, carmoisine and allura red. Further, follow-up studies to provide insight into potential adverse health effects associated with the high intakes of these artificial colour additives on the test population are warranted.     

Electrochemical sensor based on graphene and mesoporous TiO2 for the simultaneous determination of trace colourants in food

Currently, synthetic colourants draw much attention as food additives. This paper investigated the simultaneous electrocatalytic oxidation of sunset yellow and tartrazine, two yellow colourants commonly present in food together, with a novel voltammetric sensor based on graphene and mesoporous TiO₂ modified carbon paste electrode. Due to the high accumulation effect and great catalytic capability of graphene and mesoporous TiO₂, the developed sensor exhibited well-defined and separate square wave voltammetric peaks (i.e., 272 mV) for sunset yellow tartrazine. The peak currents of sunset yellow and tartrazine increased linearly with their concentration in the ranges of 0.02–2.05 μM and 0.02–1.18 μM, respectively. And the detection limit was 6.0 and 8.0 nM for sunset yellow and tartrazine, respectively. This new sensor was applied to determine sunset yellow and tartrazine in several food sample extracts. Results suggested that the proposed sensor was sensitive, rapid and reliable.     

The structural identification of a secondary dye produced from the reaction between sunset yellow and sodium metabisulphite

The stability of sunset yellow FCF (E110) was investigated in a model soft drinks system, with and without the addition of sodium metabisulphite. The rate of dye degradation was monitored by spectrophotometry and HPLC. In samples containing both sunset yellow and sodium metabisulphite, the formation of a lemon-yellow-coloured compound was clearly demonstrated. Isolation and subsequent analysis by NMR and FAB-MS showed the structure of the compound to be that resulting from sulphonation at carbon 4 of the sunset yellow molecule. Examination of a stored commercial soft drinks sample indicated the presence of the same lemon-yellow compound.     

分光光度法测定食品色素含量的研究

采用单波长法对饮料中胭脂红和亮蓝含量进行测定,结果显示,胭脂红在0.2～80mg/L,亮蓝在0.1～20mg/L浓度范围具有良好线性关系,回收率在98%～108%。采用单波长法和双波长法相结合对饮料中酒石黄和日落黄含量进行测定,结果显示,酒石黄在0.1～70mg/L,日落黄在0.2～60mg/L浓度范围具有良好线性关系,回收率在95%～110%。两种方法对样品检测快速、准确,且操作简便。利用双波长比值光谱法对果味汽水中酒石黄、日落黄、胭脂红含量进行测定,结果显示,酒石黄在0.1～60mg/L,日落黄在0.8～90mg/L,胭脂红在0.2～80mg/L浓度范围内具有良好的线性关系,平均回收率分别为100.5%、102.4%和101.8%。