分子印迹固相萃取-分光光度法检测环境水样中的孔雀石绿

目的建立分子印迹固相萃取-分光光度法检测水样中孔雀石绿的方法。方法以孔雀石绿(MG)为模板分子,甲基丙烯酸(MAA)为功能单体,乙二醇二甲基丙烯酸酯(EGDMA)为交联剂,以本体聚合方法制备的分子印迹聚合物为填料自制固相萃取柱。对固相萃取的上样溶剂、干燥方法、淋洗溶剂等条件进行了优化。采用优化的条件对自来水和河水加标水样中的孔雀石绿进行萃取,再用分光光光度计检测。结果自来水和河水水样加标浓度在8～128μg/L范围内得到的回收率分别为86.3%～92.1%和83.5%～90.3%。结论本方法操作简单、灵敏度高,适用于环境水样中孔雀石绿的痕量检测。     

分子印迹固相萃取柱-分光光度法检测 环境水样中的孔雀石绿

目的 建立分子印迹固相萃取-分光光度计检测水样中孔雀石绿的方法。方法 以孔雀石绿(MG)为模板分子, 甲基丙烯酸(MAA)为功能单体, 乙二醇二甲基丙烯酸酯(EGDMA)为交联剂, 以本体聚合方法制备的分子印迹聚合物为填料自制固相萃取柱。对固相萃取的上样溶剂、干燥方法、淋洗溶剂等条件进行了优化。采用优化的条件对自来水和河水加标水样中的孔雀石绿进行萃取, 再用分光光光度计检测。结果 自来水和河水水样加标浓度在8?128 μg/L范围内得到的回收率分别为86.3%?92.1%和83.5%?90.3%。结论 本方法操作简单、灵敏度高, 适用于环境水样中孔雀石绿的痕量检测。     

分子印迹固相微萃取技术在食品安全分析中的应用研究进展

分子印迹固相微萃取技术因具有富集效率高、选择性好、操作简便、环境友好等优点,已广泛应用于食品安全分析领域中。文章介绍了分子印迹固相微萃取介质的制备方法及包括萃取头式、管内式、整体柱式和搅拌棒式等形式在内的分子印迹固相微萃取固载技术研究进展,并综述了分子印迹固相微萃取技术在农药残留、兽药残留和有机污染物等食品安全分析领域中的应用研究进展。     

分子印迹固相萃取技术在食品农兽药残留检测中的应用进展

分子印迹固相萃取技术(英文简称MISPE),是当前食品安全样品采用的主要预处理检测技术,在实践运用中具备高效、稳定、全面等技术应用特点,受到检测机构的广泛认可和应用。本文从当前分子印迹固相萃取技术应用角度,阐述了分子印迹固相萃取原理,对分子印迹固相萃取技术在食品农兽药残留检测中的应用进行了技术分析。     

分子印迹阵列固相微萃取用于检测果蔬中多种有机磷农药残留

分别以水胺硫磷、二嗪农为模板分子,采用溶胶-凝胶法制备分子印迹固相微萃取头,以2 种萃取头各两根组装成分子印迹阵列固相微萃取装置,建立顶空固相微萃取与气相色谱联用的方法用于检测果蔬中5 种有机磷农药残留量。阵列萃取方法的检出限为0.002 8～0.043 6 μg/kg,低于单根分子印迹固相微萃取法。采用该阵列固相微萃取装置对果蔬样品中的农药残留量进行分析,苹果和马铃薯样品中5 种分析物的加标回收率均在78.7%～122.8%之间。研究表明,分子印迹阵列固相微萃取装置萃取率高、灵敏度高、准确度高、选择性好、分析通量大,适用于复杂体系中多目标物的同时分析测定。     

分子印迹固相萃取-高效液相色谱联用检测饲料中喹乙醇的研究

研究了以本体聚合法制备的喹乙醇分子印迹聚合物作为固相萃取填充材料,建立了分子印迹固相萃取与高效液相色谱联用技术检测饲料中残留痕量喹乙醇的方法。通过优化分子印迹固相萃取条件,确定用0.5ml甲醇-水(90∶10,V/V)洗脱液,在上样流速为1.2ml/min,样品在pH7~8条件下,对喹乙醇有较好富集效果。为了评价分子印迹固相萃取材料对模板分子喹乙醇的选择富集能力,分别以分子印迹聚合物和C18为固相萃取填充柱比较富集效果。在上样流速为1.2ml/min条件下,预富集42min,分子印迹固相萃取材料富集倍数达78,最低检测限(S/N=3)为88.0ng/L,连续在线富集5次的精密度(相对标准偏差)为4.6%。对添加有喹乙醇浓度为0.002 5和0.010 0mg/g的饲料进行检测,其添加回收率为89.5%和93.4%。     

分子印迹固相萃取在线检测技术在食品安全分析中的研究进展

本文介绍了固相萃取技术和分子印迹技术的基本原理,以及结合两者优点来进行分离纯化过程的技术——分子印迹固相萃取(molecularly imprinted solid phase extraction,MISPE)技术。MISPE有两种操作模式,一种为离线模式,而另一种为在线模式。相比离线模式,其在线操作不仅减少了样品前处理的时间,而且提高了分析检测的精密度、灵敏度和准确度,同时更有利于实现绿色化学。本文重点综述了近年来国内外分子印迹固相萃取在线检测技术在食品安全中的研究进展,特别是这些方法在食品中抗生素残留、农药残留、激素及非法添加物分析中的应用。同时,对分子印迹固相萃取在线检测技术所存在的问题及前景进行了展望。     

白藜芦醇分子印迹聚合物的应用及性能研究

采用分子印迹技术以白黎芦醇为模板分子,选择丙烯酰胺作为白藜芦醇印迹聚合物的功能单体,氯仿和四氢呋喃的混合溶液作为印迹聚合物合成溶剂,制备了白黎芦醇分子印迹聚合物,考察了分子印迹聚合物的吸附性能、选择性能以及固相萃取性能。优化分子印迹固相萃取的萃取条件,选择30%(体积分数)的乙醇-水溶液5 m L作为上样溶液,选择40%(体积分数)的乙醇-水溶液作为淋洗液、80%(体积分数)的乙醇-水溶液作为洗脱液。     

分子印迹固相萃取技术在食品的农药和兽药残留检测中的应用进展

分子印迹固相萃取技术是一种高效的样品前处理技术,能从复杂的样品中选择性分离目标物及其结构类似物,现已广泛应用于食品、医药、化工等多个领域。介绍了分子印迹聚合物的合成原理、方法及表征,分子印迹固相萃取的操作模式,并对近几年国内外分子印迹固相萃取技术在食品农药、兽药残留检测中的应用进行了总结。阐述了该技术目前存在的问题,并对其发展趋势进行展望,为其更好地应用于食品的分析检测提供参考。     

食品中农药残留分析前处理技术应用进展

对近年来食品中农药残留分析试样前处理技术中得到迅速发展和广泛应用的固相萃取、超临界流体萃取、基质固相分散萃取进行评述;同时对新兴的分子识别技术-分子印迹聚合物在固相萃取中的应用及影响因素加以讨论。     

超高效液相色谱-串联质谱法检测鱼中孔雀石绿、结晶紫及其代谢物

采用超高效液相色谱-串联质谱法同时测定鱼中孔雀石绿（malachite green,MG）、隐性孔雀石绿（leuco malachite green,LMG）、结晶紫（crystal violet,CV）、隐性结晶紫（leuco crystal violet,LCV）4 种物质残留。样品加入内标物D5-MG、D6-LMG、D5-CV、D6-LCV,经乙腈提取、离心、过中性氧化铝柱,氮气吹至近干,50%乙腈溶液定容,上机测定。MG和LMG线性范围为1～50 ng/mL,CV和LCV线性范围为1～100 ng/mL。4 种物质的检出限均为0.05 μg/kg,定量限均为0.15 μg/kg。在50、100、200 ng/mL 3 个添加水平下,回收率在87.80%～110.82%之间,相对标准偏差均小于等于4.32%（n=6）。该方法灵敏、准确、可靠,可以同时测定鱼中MG、LMG、CV、LCV残留。     

Validation of an LC-MS/MS method for malachite green (MG), leucomalachite green (LMG), crystal violet (CV) and leucocrystal violet (LCV) residues in fish and shrimp

A quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous analyses of malachite green (MG), crystal violet (CV) and its major metabolites, leucomalachite green (LMG) and leucocrystal violet (LCV) residues in fish and shrimp samples has been validated. Fish and shrimp samples were extracted with citrate buffer/acetonitrile, and the extracts were purified on strong cation-exchange (SCX) solid-phase extraction (SPE) cartridge. After conversion of LMG into MG using a post column oxidation reactor containing lead (IV) oxide (PbO(2)), the effluents were analysed. Residues were analysed using positive-ion electrospray ionisation (ESI). Identification and quantification of analytes were based on the ion transitions monitored by multiple reaction monitoring (MRM). Validation of the method was carried out in accordance with the Decision 2002/657/EC, which establishes criteria and procedures for the validation of methods. The following parameters were determined: decision limit (CCα), detection capability (CCβ), linearity, accuracy, precision, selectivity, specificity and matrix effect. The decision limits (CCα) for MG, LMG, CV and LCV were 0.164, 0.161, 0.248 and 0.860 μg kg(-1). The respective detection capabilities (CCβ) were 0.222, 0.218, 0.355 and 1.162 μg kg(-1). Typical recoveries (intermediate precision) in shrimp, for MG, CV, LMG and LCV for 2.0 μg kg(-1) level fortified samples using the optimised procedure were in the range 69%, 97%, 80.3% and 71.8%, respectively. The findings demonstrate the suitability of the method to detect simultaneously MG, CV and its metabolite (LMG and LCV) in fish and shrimp.     

Determination of malachite green and crystal violet in processed fish products

This paper presents analysis of malachite green (MG) and crystal violet (CV) residues in processed fish products. Samples were homogenized and extracted with ammonium acetate buffer and acetonitrile. The extracted residues were partitioned into dichloromethane, in situ oxidized to chromic forms with 2,3-dichloro-5,6-dicyano-1,4-benzoquinone, and cleaned up on neutral alumina and propylsulfonic acid cation-exchange solid-phase extraction (SPE) cartridges. MG and CV were determined at 618 and 588 nm using HPLC with a visible detector (LC–VIS) and confirmed by LC–electrospray ionization tandem mass spectrometry (LC–ESI–MS/MS). The recoveries were as follows: MG (74.8–83.8%), LMG (80.0–88.4%), CV (68.6–73.9%), and LCV (85.5–90.0%). The method modified in this study has been evaluated by application in-house to a survey of 253 processed fish products. As a result of monitoring, MG and CV were positive in one shrimp and one eel sample, respectively. Our results showed that regular monitoring of these antibiotic residues is recommended for protection of public health.     

水产品中孔雀石绿和结晶紫残留检测技术的研究进展

三苯甲烷类染料孔雀石绿和结晶紫因具有抗菌等活性,常被违法用于水产养殖业。但孔雀石绿、结晶紫及其代谢产物隐性孔雀石绿、隐性结晶紫均具有致癌性,所以水产品中这4种碱性染料的残留检测是食品安全分析的重要问题之一。本文就孔雀石绿、结晶紫及其代谢物的化学性质、危害性、使用状况、现有的前处理方法和仪器分析方法等进行了综述。由于水产品基质复杂,样品前处理尤为重要。基于固相萃取技术、Qu ECh ERS技术的高效液相色谱法、高效液相色谱-串联质谱法等方法,适合于水产品中孔雀石绿和结晶紫及其代谢物的快速同时检测,在实际检测中得到广泛应用。同位素稀释质谱法和混合模式色谱法等新型检测技术与新型净化材料将是检测孔雀石绿和结晶紫的发展新方向。     

Development of a sensitive and group‐specific polyclonal antibody‐based enzyme‐linked immunosorbent assay (ELISA) for detection of malachite green and leucomalachite green in water and fish samples

BACKGROUND: Malachite green (MG) is widely used in fishery, since it is easily adsorbed by fish during waterborne exposure and is rapidly metabolised into leucomalachite green (LMG). However, both MG and LMG are potential carcinogens, teratogens and mutagens. In this study the LMG derivative bearing an amino group on the phenyl ring was synthesised and coupled to carrier proteins. An LMG polyclonal antibody‐based enzyme‐linked immunosorbent assay (ELISA) was developed and characterised. RESULTS: The ELISA standard curve was constructed with concentrations of 0.1–100 ng mL^?1. The IC₅₀ value for nine standard curves was in the range 0.9–2.6 ng mL^?1 and the limit of detection at a signal‐to‐noise ratio of 3 was 0.02–0.10 ng mL^?1. The cross‐reactivity values of the LMG antibody with MG, crystal violet and leucocrystal violet were 95.25, 29.07 and 212.38% respectively, while less than 0.2% cross‐reactivity was found with eight other compounds. For LMG‐spiked water and fish samples, recoveries were 76.2–95.0% and the correlation coefficient of ELISA with high‐performance liquid chromatography (HPLC) was 0.9752 (n = 7). For (LMG + MG)‐spiked fish samples the results of ELISA were similar to those of HPLC. CONCLUSION: The proposed ELISA can be utilised as an analytical tool for detecting the sum of MG and LMG in water and fish muscle samples. Copyright © 2009 Society of Chemical Industry     

高分子印记固相萃取-液相色谱质谱法测定水产品中孔雀石绿、结晶紫、亮绿及其代谢产物

目的建立一种高分子印记固相萃取-液相色谱质谱联用(MISPE-HPLC/MS2)测定水产品中孔雀石绿、结晶紫、亮绿及其代谢产物的检测方法。方法样品经乙腈提取后,经中性氧化铝柱和高分子印记固相萃取柱净化,Waters Atlantis T3色谱柱(2.1 mm×150 mm,3μm)分离,乙腈和0.1%甲酸水等度洗脱,采用选择反应监测(SRM)模式进行正离子扫描,内标法定量。结果待测化合物在0.2～10μg/L范围内有很好的相关性,相关系数大于0.99,加标水平为1、2、4μg/kg,孔雀石绿、结晶紫和亮绿的平均回收率分别为93.2%～105.9%、92.7%～107.5%和60.6%～94.4%,相对标准偏差均小于12.5%,孔雀石绿、结晶紫和亮绿的检出限分别为0.02、0.03、0.03μg/kg,定量限分别为0.07、0.10、0.10μg/kg。结论本方法前处理净化效果更好、灵敏度更高,同时采用稳定性同位素稀释技术及基质匹配标准曲线,将基质抑制效应降低到最低,适用于大批量鱼类产品中孔雀石绿、结晶紫和亮绿的检测。     

同位素稀释-超高效液相色谱-串联质谱法测定水产品中孔雀石绿、结晶紫及其代谢物残留量的不确定度评定

建立同位素稀释-超高效液相色谱-串联质谱法同时测定水产品中孔雀石绿、结晶紫、隐性孔雀石绿、隐性结晶紫残留的不确定度评定方法,依据GB/T 19857-2005《水产品中孔雀石绿和结晶紫残留量的测定》,建立数学模型,通过不确定度来源分析建立不确定度评价方法。结果表明:标准溶液的配制和标准曲线拟合引入的不确定度最大,而同位素稀释法可以减小前处理过程和基质效应引入的不确定度。水产品中孔雀石绿残留量为14.26 μg/kg时,其扩展不确定度为1.35 μg/kg(k=2);结晶紫残留量为15.28 μg/kg时,其扩展不确定度为1.29 μg/kg(k=2);隐性孔雀石绿残留量为14.47 μg/kg时,其扩展不确定度为1.21 μg/kg(k=2);隐性结晶紫残留量为14.69 μg/kg时,其扩展不确定度为1.24 μg/kg(k=2)。     

Selectivity of molecularly imprinted solid phase extraction for sterol compounds

MIPs (molecularly imprinted polymers) for 17β-estradiol were prepared using 2-(trifluoromethyl) acrylic acid (TFMAA) as functional monomer and trimethylolpropane trimethacrylate (TRIM) as cross-linker by precipitation polymerisation. Data from HPLC system were calculated, and separation properties of several MIPs were reported. MISPEs (molecularly imprinted solid phase extractions) were performed with MIPs as sorbent, and absorption properties of the MISPEs prepared by UV irradiation initiating polymerisation at a ratio of 1/4 (17β-estradiol/TFMAA) for 17α-estradiol, 17β-estradiol, 4-androstene-3, 17-dione and progesterone investigated by detecting filtrates at each stage of sampling, washing and elution. Elution trials proved that addition acid or alkali reagents might affect retention of the MISPE and CSPE (control/no molecularly imprinted solid phase extraction) for 17β-estradiol. A competition test implicates that the MISPE has the strongest specific retention and enrichment for progesterone. In the actual experiment for 17β-estradiol with extracting solution of milk powder as samples, the MISPE showed better selectivity and enrichment property than C18 and CSPE, and the recovery on the MISPE was up to 85.5%.     

莠去津分子印迹聚合物微球的制备及其吸附性能与在食品检测中的应用

以莠去津为模板分子,甲基丙烯酸（methacrylic acid,MAA）为功能单体,乙腈为致孔剂,采用沉淀聚合法制备莠去津分子印迹聚合物微球（molecularly imprinted polymer microspheres,MIPMs）。对MIPMs制备工艺进行优化,当莠去津与MAA物质的量比1∶4、乙腈用量50 mL、聚合温度60 ℃时,MIPMs的吸附效果最好。通过吸附实验考察MIPMs对目标物的吸附性能,并结合Scatchard分析可知MIPMs对莠去津存在两类吸附位点,且最大表观结合量为282.69 μmol/g。以MIPMs作为固相萃取材料,制备分子印迹固相萃取（molecularly imprinted solid phase extraction,MISPE）柱,用于样品前处理,并建立莠去津-MISPE-高效液相色谱法测定食品中4 种三嗪类农药（西玛津、莠灭净、莠去津、扑草净）残留的方法。结果表明,MISPE柱对4 种三嗪类农药具有特异选择性,4 种农药的线性相关系数为0.999 1～0.999 7,检出限为0.5～5 ng/mL,平均回收率在86.2%～95.7%之间,相对标准偏差为1.98%～4.51%（n=5）。该方法能够简单、准确、高选择性地检测食品中三嗪类农药残留。     

Preparation and Evaluation of Histamine Imprinted Polymer as a Selective Sorbent in Molecularly Imprinted Solid-Phase Extraction Coupled with High Performance Liquid Chromatography Analysis in Canned Fish

A series of molecularly imprinted polymers (MIPs) were prepared against histamine. Different template/monomer ratios were applied to optimize the imprinting condition. Methacrylic acid (MAA) as a functional monomer and Chloroform as a solvent were applied in polymerization process. The binding properties of MIPs were studied in comparison with a blank non-imprinted polymer. The optimized polymer, with a histamine/MAA ratio of 1/4, was selected as a sorbent in molecularly imprinted solid-phase extraction (MISPE) of histamine from canned fish. Scatchard analysis of MIP-histamine interactions revealed two types of binding sites for MIP: high affinity (K_D?=?11.11 μM) and low affinity (K_D?=?333.3 μM). The MISPE procedure was calibrated and a recovery of 76.5–97.6 % was obtained. The intra-and inter-day precision values were less than 5.70 % and 10.1 %, respectively. The selectivity of MISPE for histamine was also studied in comparison with some other structurally similar amines, which could be simultaneously present in canned fish. The performance of the imprinted polymer was examined and the results indicated that its good selectivity and affinity for histamine was very promising. Therefore, the proposed calibrated method could be applied in selective extraction and analysis of histamine in canned fish.