凝胶净化/超高效液相色谱串联质谱法测定调味酱中32种工业染料

建立了调味酱中32种工业染料的超高效液相色谱串联质谱分析方法。样品用乙腈均质提取,经凝胶色谱净化,浓缩后,以甲醇定容,超高效液相色谱分离,串联四极杆质谱多反应监测方式监测。结果表明,32种工业染料在各自的线性范围内线性关系良好,相关系数(r)均大于0.994,定量下限为0.2～5.0μg/kg,加标回收率为53%～115%,相对标准偏差为3.9%～22.4%。该方法具有前处理简单、净化效果好、灵敏度高的优点,适用于调味酱中多组分工业染料的快速确认和定量检测。     

液相色谱/四极杆-飞行时间质谱法筛查奶酪中29种禁用和限用合成色素

建立了奶酪样品中29种禁用和限用合成色素的液相色谱/四极杆-飞行时间质谱(LC/Q-TOF MS)筛查方法。样品经正己烷-水(3:1, v/v)振荡提取,得到正己烷层、水层和残渣3部分。正己烷层经旋转蒸发浓缩后,用乙酸乙酯-环己烷(1:1, v/v)溶解,经过凝胶渗透色谱(GPC)净化去除油脂。水层经乙腈振荡提取,得到乙腈-水提取液。残渣经氨水-甲醇(1:99, v/v)溶液振荡提取,得到氨水-甲醇提取液。乙腈-水提取液和氨水-甲醇提取液不需净化直接分析。结果表明: 29种不同极性范围的合成色素化合物分别得到了有效的提取,提取回收率为70%～95%。而Q-TOF MS提供的精确质量数定性功能可以将不同种类的合成色素化合物筛查出来,各化合物与精确质量质谱库中化合物的匹配度为59.66～99.47。通过Target MS/MS扫描方式进行定量,得到8种苏丹类化合物的方法检出限为0.4～2.5 μg/kg, 21种水溶性合成色素及染料化合物的检出限为20～80 μg/kg。该方法对禁用和限用合成色素的筛查范围广泛,对含有蛋白质、脂肪等基质的食品具有较好的适用性。     

固相萃取-高效液相色谱法检测食品中的非食用色素

采用超声波提取,固相萃取柱净化,液相色谱定量检测可能在食品中非法添加的5种色素。方法对样品溶液的pH、洗脱溶剂以及洗脱体积等条件进行了优化。方法对5种色素的检出限低于81ng/mL,回收率均在87%以上,相对标准偏差(n=6)在0.65%~5.1%范围内,在0.1~100μg/mL范围内有良好的线性关系,相关系数大于0.9998。本方法可以用于食品中非食用色素的同时快速检测。     

蟹制品中合成色素的多组分同时测定方法研究

建立了高效液相色谱-二极管阵列检测器多组分同时测定蟹肉棒和蟹糊等蟹制品中柠檬黄、日落黄、苋菜红、赤藓红、偶氮玉红、胭脂红、诱惑红和红色2G等8种合成色素的方法。样品中的合成色素以甲醇-4 mol/L尿素(1+1,V/V)溶液作提取剂进行提取,提取液经聚酰胺柱净化后,在XDB-C18色谱柱上,由0.01 mol/L乙酸钠和甲醇-乙腈(1+1,V/V)组成流动相梯度洗脱下实现了8种合成色素的分离,并在各色素最大吸收波长下进行了定量检测和紫外光谱确证。应用于实际样品的检测,加标浓度分别为5 mg/kg和15 mg/kg时,回收率分别在82.5%~102.8%之间,相对标准偏差在1.0%~6.6%范围内,方法可用于蟹制品中合成色素的检测。     

液相色谱/线性离子阱-静电场轨道阱高分辨质谱法快速筛查葡萄酒中的合成色素

基于液相色谱/线性离子阱-静电场轨道阱高分辨质谱(LC/LTQ-orbitrap MS)技术构建了合成色素的质谱数据库,利用精确质量数和该数据库中二级质谱图的匹配度分析筛查,建立了葡萄酒中15种水溶性合成色素的快速筛查方法。葡萄酒样品通过弱阴离子交换柱进行净化后经苯基色谱柱分离,然后用LTQ-orbitrap MS对样品中的色素进行筛查和定量检测。结果显示,葡萄酒中15种色素的检出限在0.00040~0.18 mg/L之间,3个加标水平的回收率在43.1%~127%之间,平均相对标准偏差均小于10%。加标样品的二级质谱图与数据库中标准样品二级质谱图的匹配度均达到98%以上。该方法可在无标准物质的情况下对葡萄酒中的15种合成色素进行筛查检测。     

在线固相萃取-高效液相色谱检测饮料中5种食用合成色素

建立了同时测定饮料中5种食用合成色素的在线固相萃取-高效液相色谱-紫外检测(Online-SPE-HPLC-UV)方法。饮料样品用水稀释并经0.45μm微孔滤膜过滤后,在线固相萃取柱(Acclaim 120 C18)净化,反相C18分析柱分离,梯度洗脱,紫外检测器检测。结果表明:5种食用合成色素在0.5~20 mg/L浓度范围内线性关系良好,相关系数(r2)均大于0.999 9,检出限为0.02~0.08 mg/kg,3个水平下的加标回收率为94.5%~103.0%,相对标准偏差为0.4%~2.7%。该研究为快速准确地分离测定饮料中的食用合成色素提供了有效方法。     

鱼子酱中的合成色素反相高效液相色谱法同时测定

建立了反相高效液相色谱法同时测定鱼子酱中的柠檬黄、日落黄、偶氮玉红、苋菜红、胭脂红、赤藓红、红色2G和诱惑红8种合成色素的方法.样品中的合成色素经甲醇-4 mol/L尿素溶液(体积比1 ∶ 1)提取,固相萃取聚酰胺柱净化后,采用XDB-C18色谱柱(4.6 mm×250 mm×5 μm),以甲醇-乙腈(体积比1 ∶ 1)和0.01 mol/L乙酸钠溶液为流动相,在最佳梯度洗脱条件下,8种合成色素在17 min内实现分离.各色素在0.05 ～50 mg/L质量浓度范围内具有良好的线性关系.将该法用于实际样品的检测,8种合成色素的加标回收率为89% ～110%,相对标准偏差为1.4% ～4.7%.方法简单、高效,可用于鱼子酱中合成色素的日常监督检测.     

液相色谱-串联质谱法测定浓缩樱桃李汁中罗丹明B和苏丹红染料

建立了液相色谱-串联质谱(HPLC-MS/MS)法同时检测浓缩樱桃李汁中罗丹明B和苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ5种染料的分析方法。样品经水稀释、乙腈提取,凝胶渗透色谱(GPC)净化,采用Zorbax Eclipse Plus C18色谱柱进行分离。在电喷雾正离子模式下(ESI+),用多重反应监测(MRM)方式进行离子监测,基质匹配外标法定量。5种染料在各自线性范围内,相关系数(r2)均大于0.998,检出限(信噪比=3)为0.2~3.0μg/kg,定量限(信噪比=10)为0.5~10.0μg/kg,回收率在74.4%~95.3%之间,相对标准偏差为3.6%~7.9%。     

反相液相色谱法同时检测虾仁中合成色素的研究

建立了食品虾仁中合成色素(酸性大红GR、金黄粉、橙黄G、酸性红1号、酸性红26)的反相液相色谱同时检测方法。并且对提取剂、NH4Ac浓度和离子对种类等因素进行优化,使得5种色素完全分离,并且在色谱图上能与国标内的6种色素完全分离。金黄粉、酸性大红GR、橙黄G、酸性红1号、酸性红26的回收率均在80%以上,在0.05~10μg/mL范围内有良好的线性关系,相关系数大于0.9998。本法可用于食品虾仁中合成着色剂的快速检测。     

超高压液相色谱-飞行时间质谱法测定食品中19种非法染料

建立了同时测定食品中19种非法染料的方法。样品经丙酮/正己烷提取后,采用氧化铝柱层析净化,超高压液相色谱C18短柱分离,流动相用乙腈:0.1%甲酸溶液梯度淋洗,采用电喷雾离子源,在正离子模式下以飞行时间质谱检测,质量偏差小于1毫道尔顿。19种非法染料加标回收率在76%～108%,之间,方法的检测限为0.34～14μg/kg,精密度RSD在0.3%～12%之间。     

Establishment of an immunoaffinity chromatography for simultaneously selective extraction of Sudan I, II, III and IV from food samples

The establishment of an immunoaffinity chromatography (IAC) for simultaneously selective extraction of four illegal colorants Sudan dyes (Sudan I, II, II and IV) from food samples was described. The IAC column was constructed by covalently coupling monoclonal antibody (mAb) against Sudan I to CNBr-activated Sepharose 4B and packed into a common solid phase extraction (SPE) cartridge. It was observed that IAC column was able to separately capture Sudan I, II, III and IV with maximum capacity of 295, 156, 184 and 173ng, respectively. The extraction conditions including loading, washing and eluting solutions were carefully optimized. Under optimal conditions, the extraction recoveries of the IAC column for Sudan I-IV at two different spiked concentrations were within 95.3-106.9%. After 50 times repeated usage, 64% of the maximum capacity was still remained. Six food samples randomly collected from local supermarket without spiking Sudan dyes were extracted with IAC column and detected by high performance liquid chromatography (HPLC). It was found that there was no detectable Sudan II, III and IV in all six food samples, but Sudan I with the content of 2.7-134.5ngg(-1) was detected in three food samples. To further verify the extraction efficiency, other three negative samples were spiked with Sudan I-IV at the concentrations of 20ngg(-1) and 50ngg(-1), which were then extracted with IAC column. The extraction recoveries and relative standard deviation (RSD) were 68.6-96.0% and 4.8-15.2%, respectively, demonstrating the feasibility of the prepared IAC column for Sudan dyes extraction.     

Fast and simple method for the detection and quantification of 15 synthetic dyes in sauce,cotton candy,and pickle by liquid chromatography/tandem mass spectrometry

A simple and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the simultaneous determination of 15 illegal dyes (Sudan I, Sudan II, Sudan III, Sudan IV, Sudan Red G, Sudan Orange G, Sudan Red 7B, Para Red, Dimethyl Yellow, Rahodamine B, Sudan Black B, Sudan Red B, Auramine O, Toluidine Red and Orange II) was developed and validated in sauce, cotton candy, and pickle. The samples were extracted with acetonitrile without the use of solid-phase extraction cartridges. Chromatographic separation was achieved on a Zorbax Eclipse Plus C18 column with a flow rate of 500 µL/min at 45 °C, using a gradient elution with A (10 mM ammonium formate in water with 0.1% formic acid) and B (10 mM ammonium formate in acetonitrile (ACN) with 0.1% formic acid) as the mobile phase. The detection was performed on a AB Sciex 6500 Qtrap mass analyzer under multiple reaction monitoring mode. Limit of detection, quantification, linearity, and precision were determined during the validation process. Recoveries ranged from 82% to 119% for all synthetic dyes, in exception to Orange II in cotton candy and pickle, where signal was suppressed due to high matrix interference and poor ionization. This method offers a simple and rapid approach to detect and quantify prohibited dyes in foodstuff that can be utilized in food contaminant laboratories.     

Ionic liquids extraction of Para Red and Sudan dyes from chilli powder, chilli oil and food additive combined with high performance liquid chromatography

A simple analytical method, based on the coupling of ionic liquid-based extraction with high performance liquid chromatography (HPLC), is developed for the determination of Sudan dyes (I, II, III and IV) and Para Red in chilli powder, chilli oil and food additive samples. Two ionic liquids (ILs), 1-butyl-3-methylimidazolium hexafluorophosphate ([C₄mim][PF₆]) and 1-octyl-3-methylimidazolium hexafluorophosphate ([C₈mim][PF₆]), were compared as extraction solvents; experiments indicated that the latter possesses higher recoveries for each analyte. Parameters related to extraction of Sudan dyes and Para Red were also optimized. Under the optimal conditions, good reproducibility of extraction performance was obtained, with the relative standard deviation (RSD) values ranging from 2.0% to 3.5%. The detection limits of Sudan dyes and Para Red (LOD, S/N = 3) were in the range of 7.0-8.2 μg kg⁻¹ for chilli powder and 11.2-13.2 μg L⁻¹ for chilli oil and food additive. The recoveries were in the range of 76.8-109.5% for chilli powder samples and 70.7-107.8% for chilli oil and food additive samples.     

凝胶净化/超高效液相色谱电喷雾质谱法检测调味油中11种禁用偶氮染料及罗丹明B

建立了凝胶渗透色谱(GPC)净化/超高效液相色谱-电喷雾串联四极杆质谱(UPLC-MS/MS)同时检测调味油中11种脂溶性偶氮类工业染料(苏丹红Ⅰ、苏丹红Ⅱ、苏丹红Ⅲ、苏丹红Ⅳ、苏丹红7B、苏丹红G、苏丹黄、苏丹橙G、苏丹蓝Ⅱ、甲苯胺红、对位红)和罗丹明B的分析方法。样品经乙酸乙酯-环己烷(1∶1)提取,采用凝胶渗透色谱(GPC)去除大分子油脂、天然色素等干扰物质;以乙腈-0.1%甲酸为流动相,目标化合物在梯度洗脱条件下经C18柱分离后采用多离子反应监测(MRM)正离子模式进行检测。结果表明:调味油中12种工业染料的线性范围为1～50μg/L,定量下限(LOQ)为0.2～2.5μg/kg,在5、20、40μg/L 3个加标水平下的回收率为54%～125%,相对标准偏差为2.5%～17.2%。随机抽取市售35份样品进行检测,其中两份样品检出罗丹明B。该方法操作简便、灵敏度高,实现了11种禁用偶氮染料和罗丹明B的同时提取和净化,适合于调味油中非法添加脂溶性偶氮工业染料和罗丹明B的筛查与确证。     

Analysis of illegal dyes in food by LC/TOF-MS

A sensitive and accurate methodology was developed for the analysis of seven illegal dyes (Sudan I, Sudan II, Sudan III, Sudan IV, Sudan Orange G, Sudan R and Para Red,) used as additives in food products, such as chilli powder and steak sauces. The analytical methodology consisted of solvent extraction with acetonitrile followed by liquid chromatography time-of-flight mass spectrometry detection. Accurate mass measurements were crucial in order to achieve a high degree of specificity for the target analytes in such complex samples. The dyes were effectively extracted from spice and sauce matrices achieving recoveries higher than 75%. Because of the excellent mass accuracy obtained for the target analytes (better than 2?ppm), no cleanup of the samples was required using this methodology, thus leading to a better precision and reproducibility of the results from the quantitative point of view. Calibration curves were linear and covered two orders of magnitude (from 0.01 to 1?mg?L^?1) for all the compounds studied with the exception of Para Red. A detailed study of matrix effects is also included in this work, showing a clear improvement when dilution of the extracts was carried out. Method detection limits were in the low mg?kg^?1 range, and the precision, calculated as the relative standard deviation, ranged from 5 to 15%. The methodology was successfully applied to market samples in a survey performed as part of a regional research programme organized by the Andalusian Health Service in Spain, and a positive confirmation for Sudan I was obtained in a chilli powder sample.     

Determination of synthetic dyes in bean and meat products by liquid chromatography with tandem mass spectrometry

A sensitive and efficient method was developed for the simultaneous determination of eight synthetic dyes (Chrysoidin, Auramine O, Sudan(I–IV), Para Red, and Rhodamine B) in bean and meat products using high‐performance liquid chromatography with tandem mass spectrometry. A simple extraction procedure using acetonitrile has been applied for the extraction of these dyes from spiked bean and meat samples. Chromatographic separation was achieved on a Waters XTerra C₁₈ column (2.1 × 150 mm, 5 μm) with a multistep gradient elution. Detection and quantification were performed using mass spectrometry in multiple reaction monitoring mode. Linear calibrations were obtained with correlation coefficients R² > 0.99. The limits of detection and quantification for the eight dyes were in the ranges of 0.03–0.75 and 0.1–2.0 μg/kg depending on matrices, respectively. The recoveries of these dyes in different food matrices were between 71.2 and 116.9% with relative standard deviations <15.2%, suggesting that the developed method is promising for the accurate quantification of the eight dyes at trace levels in bean and meat products.     

Simultaneous Determination of Four Sudan Dyes in Egg Yolks by Molecularly Imprinted SPE Coupled with LC-UV Detection

A highly selective molecularly imprinted solid-phase extraction (MISPE) combined with liquid chromatography-ultraviolet detection was developed for the simultaneous isolation and determination of four Sudan dyes (I, II, III and IV) in egg-yolk products. The imprinted microspheres synthesized by suspension polymerization using phenylamine?Cnaphthol as mimic template show high selectivity and affinity to the four kinds of Sudan dyes and were successfully applied as selective sorbents of solid-phase extraction for the simultaneous determination of the four Sudans from egg-yolk samples. Good linearity was obtained in a range of 0.062?C10 ??g g^?1 and the average recoveries of the four Sudans at three spiked levels ranged from 94.1 to 102.5% with the relative standard deviations less than 5.8%. The developed extraction protocol eliminated the effect of template leakage on quantitative analysis and could be applied for the determination of Sudans in complicated food samples.     

腐殖酸键合硅胶固相萃取-液相色谱法测定辣椒粉及辣椒油中的苏丹红

以腐殖酸键合硅胶作为固相萃取介质,建立了固相萃取柱净化、高效液相色谱同时     

固相萃取-高效液相色谱法同时测定调味品中15种工业合成染料

建立了测定调味品中15种工业合成染料的固相萃取-高效液相色谱法(SPE-HPLC)。样品经甲醇-水(1:1, v/v)超声提取、SPE柱净化后用HPLC进行分析,流动相为10 mmol/L乙酸铵溶液(含1%乙酸)和乙腈。实验结果表明15种工业合成染料的分离效果良好,回收率为84.6%～114.2%,相对标准偏差为0.9%～10.3%;检出限为0.05～0.18 mg/kg。该方法操作简单,结果准确,重现性好,可用于同时测定调味品中非法添加的15种工业合成染料。