Plasticizers and BPA in spices and aromatic herbs of Mediterranean areas

Abstract

This research is carried out in order to characterize the actual contamination by two ubiquitous environmental pollutants, plasticizers and Bisphenol A, in spices (black pepper, caraway and coriander) and aromatic herbs (fennel, laurel, mint, oregano, rosemary, thyme and verbena) from Algeria (n?=?26), Tunisia (n?=?65) and Italy (n?=?53). Algerian samples seem to contain fewer residues than Italian and Tunisian samples. Among the Italian samples, only aromatic herbs, precisely mint, oregano, and laurel, were contaminated. In general, all Tunisian samples showed five plasticizers residues: the caraway, among the spices, and the rosemary, among the aromatic herbs, are found to contain more residues. Also, dietary intake of these contaminants by spices and aromatic herbs under analysis seems not to constitute a risk to the consumers.     

Mycotoxins presence in pre- and post-fermented silage from Tunisia

Silage represents a major part of the feed ration for livestock, being a potential cause of mycotoxicosis as it can be contaminated with toxigenic fungi capable of producing mycotoxins in suitable environmental conditions.In the present work, the presence of natural mycotoxins in samples collected of silage from the main producing areas in Tunisia (Ariana, Bizerte, Béjà and Jendouba) was assessed based on different plant material (oat, barley, triticale, artichoke, sulla or raygrass). Mycotoxins were evaluated during three periods in green forage (P1) and subsequently in ensiled products after 60 days (P2) and 100 days (P3) of fermentation. Samples were extracted by a QuEChERS procedure and analyzed by mass spectrometry for the determination of 23 mycotoxins. The results showed the presence of Fusarium mycotoxins, deoxynivalenol (DON), HT-2 toxin, zearalenone (ZEA), enniatins (ENA1, ENB, ENB1) and beauvericine (BEA); as well as one Alternaria mycotoxin tentoxin (TENT). The highest values detected were for DON in the three periods. DON maximum value were 381, 2053 and 916 µg/kg for P1, P2 and P3, respectively. This study demonstrated the presence of fungi and DON, ZEA and ENs contamination in Tunisian silage without a recognized risk on the ruminants or further in humans. However, a continuous enhancement of the silage quality, and management and control of mycotoxins, should be implemented to ensure safe ensilable plant material.     

基于QuEChERS提取的液相色谱-串联质谱法测定干腌火腿中15种真菌毒素

建立了干腌火腿中15种真菌毒素(黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2、赭曲霉毒素A、橘青霉素、T-2毒素、HT-2毒素、蛇形菌素、新茄镰孢菌醇、疣孢青霉原、O-甲基杂色曲霉素、杂色曲霉素、环匹阿尼酸、青霉酸)多残留的液相色谱-线性离子阱质谱(QTrap LC-MS/MS)检测方法。火腿样品经含1%甲酸的乙腈水溶液提取,Qu ECh ERS方法净化后,以含0.1%甲酸和9.9%乙腈的5 mmol/L乙酸铵水溶液(A)与含0.1%甲酸的乙腈(B)为流动相,经Eclipse Plus C18色谱柱(3.0 mm×100mm,1.8μm)分离,以多级反应监测(MRM),通过信息相关扫描方式(IDA)触发增强子离子扫描(EPI),结合建立的15种真菌毒素的标准谱图库检索的多模式进行分析。结果表明,15种真菌毒素在0.05~200μg/L范围呈良好线性,相关系数均大于0.993,定量下限为0.05~2.5μg/kg。样品在1倍、2倍、10倍定量下限3个加标水平下的平均回收率为79.1%~95.5%,相对标准偏差(RSD)为3.2%~12.8%。该方法灵敏、简便、准确,可用于干腌肉类中真菌毒素的检测分析。     

免疫亲和柱净化-液相色谱-串联质谱法测定中药材中的14种真菌毒素

依次采用磷酸盐缓冲液(PBS)和甲醇-PBS溶液提取样品,以多功能免疫亲和柱净化,采用液相色谱-串联四极杆质谱检测,可同时测定中药材中的黄曲霉毒素B1、黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2、伏马毒素B1、伏马毒素B2、T-2毒素、HT-2毒素、赭曲霉毒素A(OTA)、玉米赤霉烯酮等14种真菌毒素。优化条件下,真菌毒素的定量限(LOQ)为1～5 μg/kg, 4种中药材基质(人参、桔梗、板蓝根、麦门冬)中3个不同添加水平的平均回收率(n=6)为71.9%～99.7%,相对标准偏差(RSD)为4.8%～15.8%。该方法的检测速度快,中药材复杂基质的干扰较少,结果准确、可靠,定量限可满足国内外中药材真菌毒素相关限量的要求。     

Mycotoxin profile of Fusarium langsethiae isolated from wheat in Italy: production of type‐A trichothecenes and relevant glucosyl derivatives

Fusarium langsethiae, formally described as a new species over a decade ago, has been identified as the main producer of HT‐2 (HT2) and T‐2 (T2) toxins in Europe in small cereal grains. Mycotoxin contamination caused by this Fusarium species can represent a food safety hazard that deserves further attention. In the present work, the mycotoxin profile in wheat cultures of F. langsethiae is presented with particular reference to the production of major type‐A trichothecenes and their glucosyl derivatives. F. langsethiae isolates, representative of the major Italian wheat cultivation areas, were tested for the production of T2, HT2, diacetoxyscirpenol (DAS) and neosolaniol (NEO), and relevant glucosyl derivatives. Liquid chromatography‐tandem mass spectrometry (LC‐MS/MS) was used for the identification and chemical characterization of these metabolites. F. langsethiae isolates under investigation resulted to be potent producers of T2, HT2 and NEO. Furthermore, a well‐defined set of isolates, all originating from Central Italy, produced also DAS. All isolates were found to be able to produce HT2 glucosyl derivatives, whereas only traces of T2 glucoside were detected in one sample. Furthermore, two mono‐glucosyl derivatives of NEO and one mono‐glucoside derivative of DAS were identified and characterized. The screening for the presence/absence of glucosylated trichothecenes in analyzed fungal extracts revealed a general co‐occurrence of these derivatives with the parent toxin at levels that could be roughly estimated to account up to 37% of the relevant unconjugated toxin. This is the first report of the production of glucosylated trichothecenes by F. langsethiae cultured on small grains. Copyright © 2013 John Wiley & Sons, Ltd.     

基于QuEChERS法提取液相色谱-串联质谱法测定新会陈皮中的9种真菌毒素和农药残留

建立了Qu ECh ERS-改性多壁碳纳米管提取净化结合液相色谱-质谱联用同时检测新会陈皮中6种真菌毒素和3种农药残留的分析方法,并对影响提取、净化、检测效率的因素进行了优化。以乙腈-水(80∶20)提取样品,适量改性多壁碳纳米管净化后,净化液直接用HPLC-MS/MS进行测定,选择多反应监测模式,基质匹配标准溶液外标法定量。在优化实验条件下,9种目标化合物在各自线性范围内均具有良好的线性关系,相关系数为0.983 8~0.998 2,检出限(S/N=3)为0.18~10μg/kg。在低、中、高3个加标水平的平均回收率为72.4%~106%,相对标准偏差为2.2%~7.4%。该法准确、灵敏度高﹑操作简单﹑快速,可满足新会陈皮中上述9种化合物同时测定的要求,应用于真菌毒素和农药残留的快速筛查和确证,结果满意。     

Rapid all-in-one three-step immunoassay for non-instrumental detection of ochratoxin A in high-coloured herbs and spices

A feasible three-step method for ochratoxin A (OTA) rapid detection was developed and applied for OTA screening in high-coloured matrices such as liquorice, ginger, nutmeg, black pepper, white pepper and Capsicum spp. spices at a control level of 10 μg kg⁻¹. The method was based on the clean-up tandem immunoassay column and involved three steps: extract application, washing step and application of chromogenic substrate. A significant simplification of the assay was reached by using an additional frit with conjugate inside the clean-up tandem immunoassay column. The time for analysis was less than 10 min, including 5 min for colour development. Results were visually evaluated as colour development for negative result or no colour development for positive result. The method was coupled with a simple methanol-based extraction. A total of 27 samples were screened for OTA with the proposed method. It was shown that two samples of red pepper and one sample of liquorice, pili-pili, chilli and cayenne were contaminated with OTA above the control level at 10 μg kg⁻¹, but none of tested ginger, nutmeg, black pepper and white pepper.     

红枣中4种链格孢菌毒素的检测及风险评估

建立了超高效液相色谱-三重四极杆质谱(UPLC-MS/MS)快速检测红枣中链格孢菌代谢产生的细交链格孢菌酮酸(TEA)、链格孢酚(AOH)、交链格孢酚单甲醚(AME)和腾毒素(TEN) 4种链格孢菌毒素的方法。采用2. 0%甲酸-乙腈溶液超声提取,辅以0. 2 g Na Cl盐析,使用BEH C18色谱柱在6 min内快速分离毒素,并采用多反应监测模式(MRM)检测红枣中的4种链格孢菌毒素。除了链格孢酚的线性范围为1. 0～100μg/L外,其他3种毒素的线性范围均为10～1 000μg/L,线性关系(r2)均大于0. 990,检出限为0. 5～12. 4μg/kg。4种毒素在空白基质中的加标回收率为92. 8%～116%,相对标准偏差为0. 5%～11. 2%,可满足红枣中链格孢菌毒素的检测要求。运用毒理学软件理论计算得到成年人对4种毒素的允许每日暴露量(PDE)为19. 30～25. 65μg/d。     

Characterization of selected paper documents from the archives of Palazzo Ducale (Venice), Italy using various analytical techniques

The paper manufacturing process remained unchanged for many centuries, until, in the nineteenth century, the crisis of rags led industrialized countries to patent new production processes with different raw materials. This study deals with this period of transition by analyzing four samples stored at the archives of Superintendency Beni Architettonici per il Paesaggio e per il Patrimonio Storico Artistico ed Etnoantropologico (BAPPDAD) of Venice Lagoon, based in the Palazzo Ducale.The samples were analyzed by FT-IR (ATR), SEM-EDS, HPLC-MS/MS and Py-GC/MS. These techniques allow the characterization of various raw materials and adhesives used in various stages of paper production.These analyses have found a gradual introduction of ground wood in the composition of papers from the mid-nineteenth century, and the use of rosin as glue only in more recent documents.     

Rapid Determination of 10 Parabens in Spices by High Performance Liquid Chromatography-Mass Spectrometry

An ultra-high-performance liquid chromatography with electrospray ionization tandem mass spectrometry (UHPLC-ESI-MS/MS) method was developed for determination of ten parabens (methyl-, ethyl-, propyl-, isopropyl-, butyl-, isobutyl-, amyl-, isoamyl-, hexyl-, and heptyl-parabens) in spices. After simple preparation, the extract was injected directly into the UHPLC-MS/MS system. Parabens were separated on an Acquity UPLC BEH C18 column using gradient elution programming and detected by the tandem quadrupole mass spectrometer operating in a negative-ion electrospray-ionization (-ESI) mode. The total run time of liquid chromatography was 13 min. Linear calibration curves of 10 parabens were obtained in the range of 0.10–20.00 ng/mL under optimum conditions (r > 0.9990). The limits of detection (LOD) and the limits of quantification (LOQ) were in the range between 0.01–0.08 ng/g and 0.03–0.20 ng/g, respectively. Intra-day precision and inter-day precision were 1.04%–2.13% and 1.03%–3.16%, respectively. Recoveries were between 89.41% and 99.30%. This proposed method was successfully applied in 11 kinds of spices (38 brands). In these samples, combinations of parabens appeared more frequently than we anticipated, and 63.16% of them contained more than one paraben. As a result, it was necessary to simultaneously test for 10 parabens in spices.     

基于QuEChERS提取的超高效液相色谱-串联质谱法测定小麦粉中5种镰刀菌毒素

建立了小麦粉中恩镰孢菌素A、恩镰孢菌素A1、恩镰孢菌素B、恩镰孢菌素B1、白僵菌毒素5种镰刀菌毒素的超高效液相色谱-串联质谱(HPLC-MS/MS)分析方法。小麦粉样品采用改良的Qu ECh ERS方法进行提取,无需进一步净化,以甲醇-2 mmol/L乙酸铵为流动相梯度洗脱,经Agilent Eclipse Plus C_(18)色谱柱(2.1 mm×50 mm,1.8μm)分离,在电喷雾电离(ESI)正离子模式下采用多反应监测(MRM)进行测定,基质外标法定量。在较宽的线性范围内,5种镰刀菌毒素的相关系数(r2)均不小于0.993,方法检出限为0.3~0.8μg/kg,定量下限为0.8~2.4μg/kg。样品在1倍、2倍、10倍定量下限3个加标浓度下的平均回收率为74.0%~85.4%,相对标准偏差(RSDs)为5.6%~13.1%。采用建立的方法对市售的30批次小麦粉中5种镰刀菌毒素进行筛查,数批产品检出不同含量的镰刀菌毒素。该方法简单快速、准确、灵敏,可用于小麦粉中多种镰刀菌毒素的同时分析。     

麻黄-甘草药对配伍过程中的主要药效成分的分析

利用高效液相色谱串联质谱联用法(HPLC-MS/MS)和气相色谱质谱联用法(GC-MS)分析了麻黄与甘草药对配伍前后水煎液中主要药效成分的变化,并通过小鼠的耳廓肿胀试验考察了甘草、麻黄单煎液及药对共煎液的抗炎活性变化。分别通过HPLC法和GC-MS法对甘草与麻黄中主要化学成分,甘草酸、甘草苷、麻黄碱和甲基麻黄碱进行了定量分析,通过单煎液和药对共煎液的对比,发现麻黄与甘草配伍共煎液中麻黄碱（含伪麻黄碱）的含量增加了14.52%;甲基麻黄碱（含甲基伪麻黄碱）的含量增加了64.0%;甘草酸含量增加了13.50%;而甘草苷含量降低了19.38%。药效实验证明,甘草与麻黄配伍后抗炎作用较甘草麻黄单煎液明显增强。从而在主要成分的变化程度上揭示了甘草与麻黄配伍过程中的增效机理。     

基于分散固相萃取液相色谱-串联质谱法测定大米中8种真菌毒素

建立了大米中HT-2毒素、T-2毒素、伏马毒素B1、伏马毒素B2、玉米赤霉烯酮、赭曲霉毒素A、脱氧雪腐镰刀菌烯醇和黄曲霉毒素B1共8种真菌毒素的快速测定方法。比较了3种基于分散固相萃取原理的样品前处理方法(即Qu ECh ERS方法、EMR-lipid方法以及Dis Qu E方法)对8种真菌毒素的回收率;以提取后加标法考察大米基质中各目标物LC-MS/MS分析的基质效应。结果表明,Qu ECh ERS样品前处理方法不适于伏马毒素B1、伏马毒素B2和赭曲霉毒素A分析;而EMR-lipid样品前处理方法无法消除玉米赤霉烯酮和赭曲霉毒素A在大米中的基质效应。据此采用Dis Qu E方法并优化LC-MS/MS分析参数,一次进样监测16对离子对(每个化合物2对离子对)分析大米中的8种真菌毒素残留量。8种真菌毒素在3个添加水平下的回收率为70.0%~124.1%,相对标准偏差为0.9%~16.9%,检出限(S/N≥3)为1.2~60.0μg/kg。该方法准确、灵敏,适用于大米中多种真菌毒素的快速分析。     

Development and validation of a multiresidue method for the determination of 323 pesticide residues in dry herbs using QuEChERS method and LC-ESI-MS/MS

A multiresidue method was studied for determination of 323 pesticides representing a wide range of physicochemical properties in dry herbs (chamomile and parsley) based on QuEChERS method and LC-ESI-MS/MS analysis. In the current study, three different parameters were optimised for a higher accuracy and a lower matrix effects: extraction procedures, cleanup and matrix effect. Optimum extraction efficiency was obtained at sample hydration of 10 mL water on 2 g dry herbs and soaking time for 10 min. Use of different extraction techniques supported the use of mechanical shaker in comparison with ultrasound and handed shaking. As a way to remove interfering components from final extract, different cleanup techniques were studied: three dispersive solid phase extraction (D-SPE) (C₁₈, primary secondary amine and graphitised carbon black) have been investigated; matrix effect was reduced, but several pesticides were lost during the cleanup process. Unlike the SPE (hydrophilic lipophilic balanced polymer) produced a good recovery with all substances without expected reduction in matrix effect. Moreover, sample dilution and LC-MS/MS injection volume were studied, 3 µL injection volume was found to be the best sensitive condition without sample dilution. The developed method was validated by performing recovery tests at 0.01, 0.05 and 0.1 mg/kg, the average recoveries ranged from 70% to 120%. The reproducibility expressed as relative standard deviation (RSD %) was ≤ 20%.     

液相色谱-飞行时间质谱同时测定粮食中13种真菌毒素

建立了液相色谱-飞行时间质谱(LC-TOF MS)联用技术同时检测小麦和玉米中镰刀菌、曲霉菌和青霉菌产生的13种真菌毒素的分析方法。样品经乙腈-水-乙酸(84∶15∶1,体积比)混合溶剂提取,My-cosep 226多功能净化柱和强阴离子交换柱净化后,采用LC-TOF MS检测。在电喷雾正离子模式下,以保留时间和化合物精确分子离子质量对真菌毒素进行识别,以10 ppm为提取离子窗口进行定量。结果表明,13种真菌毒素在一定的线性范围内线性关系良好,相关系数均大于0.99,质量精确度均小于5 ppm,回收率为70%～113%,相对标准偏差为0.2%～14.5%。该方法可用于粮食中多种真菌毒素的同时测定。     

固相萃取-高效液相色谱-串联质谱法检测香辛料中罗丹明B

建立了香辛料中罗丹明B的固相萃取-高效液相色谱-串联质谱分析方法。样品经乙腈提取后离心,在提取液中加入10 mL 1%三氯乙酸溶液后,用Bond Elut Plexa PCX强阳离子固相萃取柱净化、富集,Pursuit C18色谱柱(100 mm×2.0 mm, 3 μm)分离,以0.1%甲酸水和甲醇为流动相梯度洗脱,电喷雾电离正离子模式下多反应监测(MRM)模式进行定性、定量检测。结果表明,在0.6～6 μg/L范围内的线性相关系数R2＞0.99;方法的定量限为1.2 μg/kg;添加量分别为1.197、2.992及5.985 μg/L时的加标回收率为80%～121%,相对标准偏差＜15%。同时对流动相的洗脱梯度、提取溶剂、固相萃取柱等条件进行了优化。该方法的专属性较强,基质效应较小,可用于固体香辛料产品中罗丹明B的定性、定量分析。     

气相色谱-串联质谱法快速筛查测定中药材中144种农药残留

利用气相色谱-串联质谱(GC-MS/MS)检测技术,采用QuEChERS法作为样品前处理方法,建立了能应用于11种中药材中144种农药残留的检测方法。探究了样品前处理过程中提取溶剂、缓冲盐体系、净化剂组成和用量对样品提取、净化等方面的影响,最终确定了用乙腈提取,甲苯复溶,以混合净化剂净化,过有机膜后经GC-MS/MS测定,外标法定量。144种农药在10~2000 μg/kg之间线性关系良好,相关系数(r²)>0.983;除乙酰甲胺磷、灭虫威、西玛津、克菌丹、异狄氏剂、异菌脲外,其余农药的定量限(LOQ)均低于20 μg/kg;在20、50、200 μg/kg的添加水平下,除乙酰甲胺磷、艾氏剂和双甲脒回收率偏低外,其余141种农药的平均回收率在74.3%~111.8%之间,相对标准偏差(RSD)为0.5%~14.6%。与已有的标准方法对比,此方法不仅检测结果一致,而且高效、快速,准确性好,灵敏度高,适用于中药材中144种农药残留的快速筛查与定量分析。     

超高效液相色谱-串联质谱法测定中药材中三唑类杀菌剂及三嗪类除草剂的残留量

建立了超高效液相色谱-串联质谱(UPLC-MS/MS)同时测定甘草、西洋参、三七、人参、丹参5种中药材中10种三唑类杀菌剂、18种三嗪类除草剂(包括有毒代谢物)残留量的分析方法。采用QuEChERS前处理方法,样品经1%(v/v)醋酸乙腈提取,乙二胺-N-丙基硅烷(PSA)净化处理,以Shim-pack XR-ODSII(75 mm×2.0 mm)为色谱柱,0.05%(v/v)甲酸溶液-乙腈为流动相梯度洗脱,在多反应监测(MRM)模式下测定。25种农药及其3种有毒代谢物的定量限(S/N≥10)为0.20～5.52 μg/kg;检出限(S/N≥3)为0.10～2.57 μg/kg;在各自的考察浓度范围内线性关系良好(r≥0.999);在0.20～55.2 μg/kg添加水平内,平均加标回收率为70.6%～125.7%,RSD为0.7%～14.2%。该方法样品前处理简单、快速、灵敏,可用于中药材中三唑类杀菌剂和三嗪类除草剂农药残留量的快速筛查。     

A surprising method for green extraction of essential oil from dry spices: Microwave dry-diffusion and gravity

Without adding any solvent or water, we proposed a novel and green approach for the extraction of secondary metabolites from dried plant materials. This "solvent, water and vapor free" approach based on a simple principle involves the application of microwave irradiation and earth gravity to extract the essential oil from dried caraway seeds. Microwave dry-diffusion and gravity (MDG) has been compared with a conventional technique, hydrodistillation (HD), for the extraction of essential oil from dried caraway seeds. Essential oils isolated by MDG were quantitatively (yield) and qualitatively (aromatic profile) similar to those obtained by HD, but MDG was better than HD in terms of rapidity (45min versus 300min), energy saving, and cleanliness. The present apparatus permits fast and efficient extraction, reduces waste, avoids water and solvent consumption, and allows substantial energy savings.     

高效液相色谱-质谱/质谱测定血浆中格列苯脲

本文建立了人血浆中格列苯脲的高效液相色谱-质谱/质谱(HPLC-MS/MS)分析方法。血浆中格列苯脲测定的线性范围是1~300 ng/mL;高、中、低三个浓度分析测定的精密度(RSD)<15%;格列苯脲和格列齐特(内标)绝对回收率均>80%。本方法灵敏、准确,重现性好,用于血药浓度的测定获得满意结果。