HPLC-PDA法测定食品中微量苏丹红Ⅲ

建立了利用高效液相色谱(HPLC)-二极管阵列检测器(PDA)法测定食品中苏丹红Ⅲ的方法。采用的色谱条件为:Symmetry C_(18)色谱柱(4.6 mm×250 mm,5μm),用乙腈-丙酮(80-20):水=95∶5作为流动相,检测波长为504 nm,流动相流速为1.0 m L/min,柱温为15℃。利用色谱检测苏丹红Ⅲ的浓度在0.03μg/m L~101.86μg/m L时,组分的峰面积与浓度之间线性关系良好,苏丹红Ⅲ的线性方程为A=26 310ρ-911.78(ρ:μg/m L),R~2=0.999 2。该方法精密度良好,稳定性较强。检测的4种样品的加标回收率在97.33%~100.00%之间,符合测定要求。高效液相色谱法,方法准确,快速,适用于苏丹红Ⅲ的测定。通过质谱,进一步验证了食品中苏丹红Ⅲ的存在。利用质谱检测到,苏丹红Ⅲ的分子离子峰为m/z 353,主要碎裂成m/z 156和m/z 197。     

高效液相色谱法测定食品中苏丹红I~IV

目的 建立准确可靠的高效液相色谱测定食品中苏丹红Ⅰ~Ⅳ的方法。方法 用乙腈提取样品中的苏丹红,经漩涡超声离心后,将乙腈层倾倒出;用乙腈重复提取,合并乙腈层,旋转蒸发至干。用乙腈转移至凝胶色谱进样小瓶,通过凝胶色谱的净化,用液相色谱进行检测。其中凝胶色谱柱为Bio-BeadsTM S-X3 Beads,液相色谱柱为Waters C18 （4.6mm × 150mm × 5μm）,液相色谱流动相为乙腈-水（v/v,90:10）。结果 4种苏丹红染料的检出限均能达到10 μg/kg,满足国标要求。标准加入量为0.16μg/mL浓度时辣椒粉的回收率为92.5~97.5、辣椒酱的回收率为83.8~87.5、香肠的回收率为82.5~98.8,相关系数0.9995以上,且重复性良好。结论 高效液相色谱法可以测定食品中苏丹红Ⅰ~Ⅳ,适用于批量样品的检测。     

凝胶渗透色谱-液相色谱法测定辣椒及其制品中对位红和苏丹红

建立了一种凝胶渗透色谱-高效液相色谱同时检测辣椒及其制品中对位红和五种苏丹红的方法。样品经环己烷-乙酸乙酯(体积比为1:1)提取,凝胶渗透色谱(GPC)净化,采用Diamond柱(250mm×4.6mmi.d.,5μm)分离,以乙腈:水(体积比为95:5)为流动相,流速1.0ml/min;用紫外检测器检测,检测波长500nm。对位红和苏丹红0.01～10.0mg/L时线性关系良好,加标回收率为90%～102%,相对标准偏差为1.6%～5.2%,六种染料的检出限均为0.01mg/kg。     

超高效液相色谱串联质谱法同时检测禽蛋中的角黄素、对位红和苏丹红Ⅰ~Ⅳ

目的 建立超高效液相色谱串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)同时测定禽蛋中角黄素、对位红和苏丹红残留量的方法。方法 禽蛋样品用乙腈超声提取, 加入氯化钠进行盐析脱水, 提取液加等体积水混合后经C18固相萃取小柱净化, 采用Waters C18色谱柱(50 mm×2.1 mm, 1.7 μm)以0.1%甲酸水和乙腈溶液为流动相梯洗脱分离, 电喷雾电离(electrospray ionization, ESI)正离子多反应监测模式进行定量分析。结果 角黄素、对位红和苏丹红质量浓度在0.2~ 100.0 μg/L范围内线性关系良好, 相关系数均在0.999以上; 在低、中、高 3个加标水平的平均回收率为80.5%~106.2%, 相对标准偏差(relative standard deviation, RSD)为0.68%~7.15%; 角黄素、对位红和苏丹红的检出限为0.2 μg/kg。结论 该方法操作简单、准确, 回收率较好, 方法检出限较低, 可应用于禽蛋中角黄素、对位红和苏丹红Ⅰ~Ⅳ的检测。     

高效液相色谱法测定棕榈油中苏丹红、对位红

样品经己烷溶解,再用乙腈萃取浓缩,经乙腈 异丙醇(1∶1)溶解过滤后,用反相高效液相色谱法-DAD检测器进行检测。方法最低检测限:苏丹红Ⅰ、苏丹红Ⅱ、苏丹红Ⅲ、苏丹红Ⅳ和对位红检测限均为10μg/kg,相对标准偏差为1.85%～5.74%,回收率为93.42%～108.38%。该方法简便、快速,稳定可靠。     

固相分散萃取-高效液相色谱法测定郫县豆瓣中四种苏丹红染料的研究

建立了郫县豆瓣中苏丹红Ⅰ～Ⅳ的固相分散萃取(SPDE)-高效液相色谱(HPLC)分析方法。样品用无水硫酸钠作为分散剂,以乙腈提取样品中的苏丹红,提取液用中性氧化铝层析柱进行净化。用Inertsil ODS-sp C18柱(4.6mm×250mm,5μm))分离,流动相A为乙腈,流动相B为0.1%甲酸水溶液(A∶B为90∶10,V/V),等度洗脱,流速1mL/min,柱温40℃。二极管阵列检测器检测,检测波长为517nm,利用保留时间和光谱图定性,外标法定量。4种苏丹红染料在0.10～20.00μg/mL范围内线性关系良好,相关系数均大于0.9999,苏丹红Ⅰ～Ⅳ的检出限(LOD)分别为0.009～0.013mg/kg(信噪比S/N=3)。在添加浓度为0.5～10.0mg/kg范围内平均回收率达81.67%～93.28%,相对标准偏差(RSD)为1.07%～4.61%(n=6)。     

高效液相色谱法测定食品中苏丹红Ⅰ~Ⅳ

目的建立准确可靠的高效液相色谱法测定食品中苏丹红I~IV的方法。方法用乙腈提取样品中的苏丹红,经漩涡超声离心后,将乙腈层倾倒出,用乙腈重复提取,通过凝胶色谱的净化,用液相色谱进行检测。其中凝胶色谱柱为Bio-BeadsTM S-X3 Beads,液相色谱柱为Waters C18(4.6 mm×150 mm,5μm),液相色谱流动相为乙腈-水(V:V,90:10)。结果 4种苏丹红染料的检出限均能达到10μg/kg,满足国标要求。标准加入量为0.16μg/m L浓度时,辣椒粉的回收率为92.5%~97.5%、辣椒酱的回收率为83.8%~87.5%、香肠的回收率为82.5%~98.8%,相关系数0.9995以上,且重复性良好。结论高效液相色谱法可以测定食品中苏丹红I~IV,适用于批量样品的检测。     

凝胶渗透-高效液相色谱法测定棕榈油中的对位红和苏丹色素

采用凝胶渗透色谱(GPC)分离棕榈油中的对位红和苏丹色素,浓缩后用纯乙腈淋洗的高效液相色谱双波长检测法测定其中各待测组分的含量.测定结果表明最低检测限:对位红、苏丹Ⅰ、Ⅱ、Ⅲ、Ⅳ分别为0.03、0.06、0.10、0.08、0.10 mg/kg;相对标准偏差为2.15%～5.23%,平均回收率为83%～95%.操作方法简便,结果准确可靠.     

超高效液相色谱-质谱法测定禽蛋中苏丹红

建立了一种快速测定禽蛋中苏丹红含量的超高效液相色谱-串联四极杆质谱联用(UPLE-MS/MS)分析方法.样品经乙腈提取后,加水反沉淀除去蛋白质和脂肪等杂质,冷冻后取上层清液,经C18>超高效液相色谱柱分离,串联四极杆质谱多反应监测模式检测.苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ方法检出限为5 μg/kg,相关系数是0.9999,在100～300μg/kg范围内,回收率在70.1%～107%之间.     

高效液相色谱法测定红酵母细胞中的β-胡萝卜素

采用反相高效液相色谱法(RP-HPLC)测定红酵母细胞中β-胡萝卜素的含量。发酵法得到的样品经过研磨,以丙酮-石油醚(1∶1 v/v)组成的溶剂提取β-胡萝卜素,经0.22μm(有机相)微孔过滤膜过滤净化,用RP-HPLC进行定量分析。色谱条件:分析柱为Hypersil RP-ODS C18(5μm,250mm×4.6mm i.d.);流动相:V(乙腈):V(四氢呋喃)=60∶40的溶液;检测波长451nm;以峰面积积分法定量。当β-胡萝卜素的质量浓度在2～18μg/mL范围时,其浓度与其峰面积之间有良好的线性关系(r=0.9971)。验证结果表明,该方法测定结果准确,能用来测定红酵母中的β-胡萝卜素。     

UPLC-ESI-MS/MS analysis of Sudan dyes and Para Red in food

An analytical method for the simultaneous determination of Sudan dyes (Sudan Red G, Sudan I, Sudan II, Sudan III, Sudan Red 7B and Sudan IV) and Para Red in food by ultra-performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-ESI-MS/MS) was developed. Samples were extracted with acetonitrile, and water added into the extract. The supernatant was analysed by UPLC-MS/MS after refrigeration and centrifugation. The sample was separated on an Acquity BEH C₁₈ column, and detected by MS/MS with the multiple reaction monitoring mode. Matrix calibration was used for quantitative testing of the method. The linear matrix calibrations of Sudan dyes and Para Red were 2–50 and 10–250 ng g^?1, respectively, and the regression coefficients were >0.9945. The recoveries were 83.4–112.3% with good coefficients of variation of 2.0–10.8%. The limits of detection were between 0.3 and 1.4 ng g^?1 for the six Sudan dyes, and between 3.7 and 6.0 ng g^?1 for Para Red. The limits of quantification were between 0.9 and 4.8 ng g^?1 for the six Sudan dyes, and between 12.2 and 19.8 ng g^?1 for Para Red.     

A rapid HPLC method for determination of Sudan dyes and Para Red in red chilli pepper

A rapid high-performance liquid chromatography (HPLC) system consisting of an ultraviolet-visible (UV–VIS) detector was developed for the separation and determination of Sudan dyes (I, II, III, and IV) and Para Red in red chilli peppers. The chromatographic separation was achieved on a reverse phase C₁₈ column with isocratic elution, using a mobile phase of acetonitrile/methanol (80:20, v/v); detector was set at 506 nm. All four Sudan dyes and Para Red were separated in less than 9 min. Among 80 red chilli peppers screened, only one of them contained 0.10, 0.04, and 0.05 mg/kg Sudans I, III, and IV, respectively. No Sudan II and Para Red were detected in any of the red chilli peppers analysed. The method was ‘in-house’ validated using red chilli peppers based on following criteria: limit of detection (LOD), limit of quantification (LOQ), recovery, repeatability, reproducibility, and linearity in red chilli peppers. Depending on the dye involved, LOD and LOQ were in the range of 1.2–5.4 and 4–18 μg/kg in red chilli, respectively. The recovery, repeatability (expressed as coefficient of variation, CV_r), and reproducibility (CV_R) varied from 89 to 98%, from 0.82 to 4.09%, and from 1.33 to 4.65%, respectively. Linearity obtained for all dyes and Para Red were all r² > 0.9999 (in the range of 0.01–5 mg/l). The applicability of the method to the determination of Sudan dyes and Para Red in red chilli peppers was demonstrated. This method has potential to be used for illegal Sudan dyes and Para Red in red chilli peppers and some foodstuffs due to its simple, reliable, rapid, and excellent precision.     

三种专用固相萃取柱-高效液相色谱法检测食品中苏丹红含量

建立了专用固相萃取柱-高效液相色谱法检测食品中苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ的方法。样品经过正己烷提取,通过苏丹红专用固相萃取柱净化和富集,40℃水浴旋蒸至干后用甲基叔丁基醚-甲醇(体积比4:6)溶解定容,借助Waters Symmetry C₁₈色谱柱(150 mm×4.6 mm,5 μm)分离,以乙腈-水为流动相梯度洗脱,二极管阵列检测器检测,外标法定量。结果表明,四种苏丹红在0.16~2.56 μg/mL内线性相关系数(r)均大于0.9999,不同食品的检出限在2.3~9.7 mg/kg之间。不同品牌的苏丹红专用固相萃取柱去除基质干扰和富集目标物的能力不同,可根据食品种类选择合适的固相萃取柱。ProElut SDH SPE柱普遍适用于不同种类食品的前处理;CNW Poly-sery MIP-SDR SPE柱适用于除辣椒粉以外的大部分食品的前处理;Cleanert Sudan SPE柱适用于浅色、低油脂食品的前处理。对六种食品加标2.0 mg/kg,经过ProElut SDH SPE柱处理后,回收率为83.7%~91.1%,相对标准偏差为2.4%~6.2%(n=6)。该方法净化和富集效果理想,与GB/T 19681-2005相比,具有操作简便、重复性好、准确度高、分析时间短、节省溶剂等特点。     

Determination of dyes sudans in of raw foods

The number of cases using of forbidden dyes (such as Sudan-I and Para Red) in European foodstuff (including foodstuff from Russia) were increased last time. In this case it was required to adapt existing analytical methods for the control of dyes Sudan I-IV and Para Red. It was used for this purpose high performance liquid chromatography with detectings on diode-array and MS detector. Sudan I and Para Red were detected in 3 foodstuffs from examined 49 products. Concentration levels of these dyes were varied from 3 up to 500 mg/kg. Limits of determination of the methods is: HPLC-diode-array--0.1 mg/kg and HPLC-MS--0.5 mg/kg.     

Discovery and structural elucidation of the illegal azo dye Basic Red 46 in sumac spice

An unknown red dye was discovered in a sumac spice sample during routine analysis for Sudan dyes. LC-DAD and LC-MS/MS did not reveal the identity of the red substance. Nevertheless, using LC-high-resolution MS and isotope ratio comparisons the structure was identified as Basic Red 46. The identity of the dye was further confirmed by comparison with a commercial hair-staining product and two textile dye formulations containing Basic Red 46. Analogous to the Sudan dyes, Basic Red 46 is an azo dye. However, some of the sample clean-up methodology utilised for the analysis of Sudan dyes in food prevents its successful detection. In contrast to the Sudan dyes, Basic Red 46 is a cation. Its cationic properties make it bind strongly to gel permeation columns and silica solid-phase extraction cartridges and prevent elution with standard eluents. This is the first report of Basic Red 46 in food. The structure elucidation of this compound as well as the disadvantages of analytical methods focusing on a narrow group of targeted analytes are discussed.     

Discovery and structural elucidation of the illegal azo dye Basic Red 46 in sumac spice

An unknown red dye was discovered in a sumac spice sample during routine analysis for Sudan dyes. LC-DAD and LC-MS/MS did not reveal the identity of the red substance. Nevertheless, using LC-high-resolution MS and isotope ratio comparisons the structure was identified as Basic Red 46. The identity of the dye was further confirmed by comparison with a commercial hair-staining product and two textile dye formulations containing Basic Red 46. Analogous to the Sudan dyes, Basic Red 46 is an azo dye. However, some of the sample clean-up methodology utilised for the analysis of Sudan dyes in food prevents its successful detection. In contrast to the Sudan dyes, Basic Red 46 is a cation. Its cationic properties make it bind strongly to gel permeation columns and silica solid-phase extraction cartridges and prevent elution with standard eluents. This is the first report of Basic Red 46 in food. The structure elucidation of this compound as well as the disadvantages of analytical methods focusing on a narrow group of targeted analytes are discussed.     

Determination of Trace Para Red Residues in Foods through On-Line Molecularly Imprinted Solid Phase Extraction Coupled with High-Performance Liquid Chromatography

ABSTRACT:  In this article, we prepared a novel imprinted polymer by a room temperature ionic liquid-mediated surface molecular imprinting technique in combination with a sol–gel process. This polymer was characterized by static and kinetic adsorption experiments and exhibited good recognition ability and offered fast kinetics for the adsorption of Para Red. A simple and sensitive analytical method, based on the coupling of molecularly imprinted solid phase extraction with high-performance liquid chromatography (HPLC), had been developed for determination of trace Para Red. With a loading flow rate of 0.42 mL min⁻¹ for 25 mL, an enrichment factor of 1061 was achieved. Under the selected experimental condition, the detection limit (S/N = 3) of Para Red was 6.6 ng L⁻¹, and the peak area precision (RSD) for 5 replicate detections of 0.15 μg L⁻¹ Para Red was 4.1%. The applicability of this method for determination of the blank chili sauce sample, spiked with Para Red at 5 to 25 ng g⁻¹ levels, was demonstrated, with recoveries ranging from 86% to 95%.
Practical Application: In this paper, a simple and sensitive analytical method, based on the coupling of molecularly imprinted solid phase extraction with high performance liquid chromatography, had been developed for determination of trace Para Red. It was applied to the analysis of spiking Para Red in chili sauce sample with satisfactory recovery and repeatability. This proposed method has the potential to be used for monitoring the illegal addition of Para Red in foods in the future due to its simple, reliable, rapid, and excellent precision.     

构建氯化血红素/G-四链体DNA酶比色生物传感器检测食品中苏丹红

以苏丹红III核酸适配体为识别元件,以氯化血红素/G-四链体DNA酶为信号探针,结合杂交链反应信号放大策略,构建一种简单、新颖、高灵敏的生物传感器,用于苏丹红比色检测。在优化条件下,对该方法的灵敏度、准确性、特异性进行评估,最后将其应用食品中苏丹红III快速检测,并与GB/T 19681—2005法对比验证。结果显示,在优化条件下,苏丹红III质量浓度在0.5～250 ng/mL范围与450 nm波长处的吸光度呈良好线性关系（相关系数为0.995）,方法检测限为0.09 ng/mL。特异性分析显示,本方法可用于苏丹红I～IV的检测。实际样品分析中表明,食品中苏丹红III加标回收率为84.3%～101.6%,相对标准偏差为4.13%～8.36%,本方法的加标回收率与GB 19681—2005法相比差异不显著（P＞0.05）。该方法操作简便、准确可靠、灵敏度高,适用于批量食品中苏丹红的快速检测。     

高效液相色谱法测定食品中可能掺杂的16 种工业染料

建立食品中16 种工业染料的高效液相色谱检测法,包括分散红11、分散黄9、分散橙3、分散橙11、分散红1、分散红9、分散黄54、分散橙37、分散橙61、苏丹红1、分散黄7、分散橙149、苏丹红2、苏丹红3、苏丹红7B、苏丹红4。选择糖果、巧克力、番茄酱与番茄沙司为基质,使用Agilent ZORBAX SB-C18色谱柱进行分离,流速为0.8 mL/min,流动相为乙腈和水,验证过程中确定检出限为0.26～0.88 mg/kg、定量限为0.82～2.52 mg/kg,16 种染料的回收率在81.0%～110.3%之间,相对标准偏差在0.3%～7.5%之间。表明方法稳定可靠,可用于日常的非法添加染料的测定和筛查。     

基质固相分散-高效液相色谱法测定川味香肠中的苏丹红染料

建立了川味香肠中苏丹红I～IV的基质固相分散(MSPD)-高效液相色谱(HPLC)分析方法。样品以无水硫酸钠作为分散基体,研磨均匀后与中性氧化铝同时装柱,正己烷淋洗净化,以丙酮-正己烷(5∶95,V/V)溶液洗脱。用InertsilODS-spC18柱(4.6mm×250mm,5μm)进行分离,流动相A为含0.1%甲酸的甲醇,流动相B为0.02mol/L的乙酸铵溶液(A∶B=85∶15,V/V),等度洗脱,柱温40℃。二极管阵列检测器检测,检测波长为490nm,利用保留时间和光谱图定性,外标法定量。4种苏丹红染料在0.10～50.00μg/mL范围内线性关系良好,相关系数均大于0.9999,苏丹红I、II、III、IV的检出限(LOD)分别为0.008～0.011mg/kg(信噪比S/N=3)。在添加浓度为5.0～25.0mg/kg范围内平均回收率达85.54%～94.66%,相对标准偏差(RSD)为0.87%～4.23%(n=6)。