NL2020667B1 - Method for improved detection of dioxins - Google Patents

Method for improved detection of dioxins Download PDF

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NL2020667B1
NL2020667B1 NL2020667A NL2020667A NL2020667B1 NL 2020667 B1 NL2020667 B1 NL 2020667B1 NL 2020667 A NL2020667 A NL 2020667A NL 2020667 A NL2020667 A NL 2020667A NL 2020667 B1 NL2020667 B1 NL 2020667B1
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dioxin
emulsion
polar phase
hexane
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Broer Wilhelmus
Markesteijn Jeroen
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Nofalab B V
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/44—Sample treatment involving radiation, e.g. heat
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/34—Purifying; Cleaning
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4044—Concentrating samples by chemical techniques; Digestion; Chemical decomposition
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/405—Concentrating samples by adsorption or absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4005—Concentrating samples by transferring a selected component through a membrane
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/40—Concentrating samples
    • G01N1/4055—Concentrating samples by solubility techniques
    • G01N2001/4061—Solvent extraction
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/08—Preparation using an enricher
    • G01N2030/085—Preparation using an enricher using absorbing precolumn

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  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Sampling And Sample Adjustment (AREA)

Abstract

De onderhavige uitvinding heeft betrekking op een werkwijze voor het vaststellen van de aanwezigheid van dioxine in een voedselproduct. De werkwijze van de onderhavige uitvinding is in het bijzonder geschikt voor detectie van dioxinen in producten die doorgaans moeilijk te analyseren zijn, onder andere producten met een hoog eiwitgehalte zoals vlees en zuivelproducten en vis- en schaaldieren.

Description

Description
The present invention relates to a method for establishing the presence of dioxin in a food product. The method of present invention is especially suitable for detection of dioxins in product having a high protein content.
Dioxins £ire a group of chemically-related compounds ( 200+ compounds) that are persistent environmental pollutants. Dioxin is a colorless, odorless organic compound containing carbon, hydrogen, oxygen and chlorine and include polychlorinated dibenzo-p-dioxins (PCDDs), derivatives of dibenzo-p-dioxin, the most toxic being 2,3,7,8-Tetrachlorodibenzodioxin (TCDD), Polychlorinated dibenzofurans (PCDFs), or furans and polychlorinated/polybrominated biphenyls (PCBs/PBBs). Under certain conditions PCBs may form dibenzofurans/dioxins through partial oxidation.
Dioxins are unwanted side products of a wide range of manufacturing processes including the manufacturing of some herbicides and pesticides. In the environment, dioxins lend to accumulate in the food chain. The contamination can vary widely depending on the origin of the foodstuff. Meat, eggs, milk, seafood and other food products may be contaminated above background levels by dioxins. The majority of human exposure to dioxins is through food, mainly meal and dairy products, fish and shellfish.
Once dioxins enter the body (e.g. via food), they can last for a long time because of their chemical stability and their ability to be absorbed by fat tissue and then stored in the body. Dioxins are highly toxic and can cause reproductive and developmental problems, damage the immune system, interfere with hormones and also cause cancer. Dioxin have been found at toxic at levels in the part per trillion (ppt) range. Therefore it is essential that dioxins are monitored the food supply, to ensure food safety. Food and feed contamination monitoring systems must be in place to ensure that tolerance levels are not exceeded. The level of detection needed for dioxins make the current methods for the analysis of these compounds difficult to perform and expensive.
Most methods for the detection of dioxins make use of isotopic dilution technique where a known amount of the carbon-13 form (internal standard) of a compound is spiked into the sample, providing a solution with chemically identical forms of the compound that can be separately quantified. The method consist of the extraction (separation/isolalion of the dioxin compounds) of the sample, and the instrumental analysis, being gas chromatograpy (GC) and mass spectrometry (MS).
The major challenges with regard to the analysis of food products lie in the extraction and separation of the dioxins and PCBs from the other compounds found in the matrix. During the process of preparing the samples for GC and MS measurement, proteins, sugars, and fat residues interfere with the detection of dioxins, therefore the sample needs extensive and time consuming clean up. Furthermore, there are significant differences in the approach to extraction and clean-up needed for fruits, hard vegetables, soft vegetables, grains, dairy products, fish and meats. However, the more complex the extraction and clean-up becomes, the greater the potential for loss of materials of interest. For instance, food product that are high in protein content remain a difficult group of food product to be analysed for their dioxin content due to the high eontamination/pollution of the matrix, resulting in a reduced sensitivity and reliability of the measurement at levels as low as at the ppt level. An alternative to the exclusion of dioxins and PCBs from protein rich products is the traditional soxhlet extraction, but this uses a lot of solvent (> 250 ml dichloromethane / n-Hexane) and is very time-consuming (~ 10 hours extration time). Furthermore, foods that combine ingredients into a final product can further complicate the matrix with the chemical additives. Finally, determination of false-negative samples are also a problem, because this includes an increased potential for contaminated products to reach the consumer. This may for instance occur when a sample matrix results in analyte loss during extraction and cleanup, the contaminant will not be detected.
Present techniques and methods for the quantitative analysis of dioxins require expensive, elaborate and complex methods. The analysis costs are high and may also vary according to the type of sample or dioxin. Considering the above, there is a need in the art for a simple level, less expensive but more reliable dioxin screening method for food and feed samples.
It is an object of the present invention, amongst other objects, to address the above need in the art. The object of present invention, amongst other objects, is met by the present invention as outlined in the appended claims.
Specifically, the above object, amongst other objects, is met, according to a first aspect, by the present invention by a method for establishing the presence of dioxin in a food product, comprising the steps of
a) contacting of a sample of the food product having a protein content of at least 15 wt% , based on the total weight of the sample, with a solution comprising sulphuric acid (H2SO4),
b) heating of the sample contacted with the sulphuric acid solution of step a to at least 100 °C for at least 30 min, preferably 1 hour, to obtain an emulsion,
c) allowing the emulsion to cool to a temperature between 15 and 25 °C,
d) contacting the emulsion of step c with an extraction liquid comprised of at least 15 wt% hexane and at least 75 wt% isopropanol, based on the total weight of the extraction liquid,
e) homogenizing of the emulsion of step d to form a suspension.
f) allowing separation of the suspension in at least two phases and subsequently isolating the non-polar phase after extraction,
g) contacting the isolated non-polar phase with hexane,
h) repeating step f) and step g) for at least one more time, to obtain a sample A,
j) subjecting sample A to a purification column comprised of AgNO3 silica gel for purification of the sample A to obtain a sample B,
k) subjecting the sample B to gas chromatography and/or mass spectrometry for establishing the presence of dioxin.
After separation in step f is allowed, two phases are formed, resulting in one layer on top of the other. The upper or top layer is a non-polar layer is comprised of hexane including the non-polar components such as dioxins and PCBs. The lower layer is the polar layer comprising all polar component in a aqueous fraction. The upper layer is continued to step g for further extraction of the dioxins and PCBs.
The method of present invention improves the extraction of dioxins, for example, by means of the speed extractor. Solid products are normally run with the speed extractor to extract the dioxins from the product before it goes to the clean-up system, but for some samples having a high protein content this does not w'ork. Samples with a protein content higher than 1520% (feather meal, fish meal, blood meal etc.) Cannot be processed with the aid of the speed extractor because the extract ensures that the tubing become clogged when they go from the extraction tube to the collection reservoir.
The method of present invention is especially suitable for food products that are high in protein content. Due to the matrix effects of the protein on the detection of dioxins, these type of product is difficult to achieve a robust and accurate detection. The method of present invention solve this problem by a specific pre-treatment of the samples. The sample contacted with the sulphuric acid solution is heated up to at least 100 0 C. The increased temperature results in an improved and accelerated break dowm of protein content present in the food samples. Especially food samples having a high protein content, i.e. a protein content of at least 15 wt% have proven to benefit from the heating step w'ith sulphuric acid. Preferably, in addition to the heating during the extraction, the product and solvent can be under high pressure, at least 100 bar. The proteins will dissolve under this temperature and pressure. However, if the solution is cooled back to room temperature, the proteins will come out of the solution again and will result in clogging of the tubing of the GC/MS. This results in low' recoveries (<50%), or no extract at all. Experiments resulted in that for products with high protein concentrations (feather meal, fishmeal, blood meal, etc.), it is preferred to perform a protein hydrolysis with a sulfuric acid solution. As a result, the proteins are broken down into amino acids and polypeptides. After this step, a liquid / liquid extraction can be performed to extract the dioxins and PCBs from the amino acid / peptide emulsion. The recoveries found by using this method are significantly higher than the first method, namely at least 70% or higher. Furthermore, besides the fact that high recovery percentages are obtained, chlorinated solvents are not used and less organic solvent is used (maximum 100ml nhexane).
According to a preferred embodiment, the present invention relates to the method wherein after step f), in addition to the non-polar phase, a polar phase is obtained and wherein said polar phase is subjected to a purification column comprised of H2SO4 silica gel and eluted with hexane to obtain a sample C. In the event that in the top phase (non-polar phase) no dioxins could have been established, the dioxins may have been ended up in the “waste fraction” i.e. the polar phase after extraction. Therefore the polar phase is to be analysed for dioxins. But before this analysis can be performed, this phase needs to be purified.
According to another preferred embodiment, the present invention relates to the method wherein the polar phase is subjected to a purification column comprised of a glass fibre membrane to obtain sample C, followed by subjecting the sample C to gas chromatography and/or mass spectrometry for establishing the presence of dioxin.
According to yet another preferred embodiment, the present invention relates to the method wherein in step a), the weight ratio between sample and sulphuric acid solution is at least 1:12, preferably at least 1:16, more preferably at least 1: 20, most preferably 1:24.
According to a preferred embodiment, the present invention relates to the method wherein the sulphuric acid solution is at least a 5M solution, more preferably at least a 6M solution.
According to another preferred embodiment, the present invention relates to the method wherein in step d the volume ratio of said extraction liquid in relation to said emulsion is at least 1:1, more preferably at least 5:1.
According to yet another preferred embodiment, the present invention relates to the method wherein in step g the volume ratio of hexane in relation to said non-polar phase is at least 1:1, more preferably at least 5:1.
According to a preferred embodiment, the present invention relates to the method wherein said dioxin is one or more selected from the group consisting of polychlorinated dibenzop-dioxins (PCDDs), derivatives of dibenzo-p-dioxin, 2,3,7,8-Tetrachlorodibenzodioxin (TCDD), polychlorinated dibenzofurans (PCDFs), furans and polychlorinated/polybrominated biphenyls (PCBs/PBBs).
According to another preferred embodiment, the present invention relates to the method wherein said food product is one or more selected from the group consisting of meat and dairy products, fish, shellfish, fruit, vegetables, seeds, grain, herbs, spices, eggs and milk, preferably meat and dairy products, fish and shellfish.
According to yet another preferred embodiment, the present invention relates to the method wherein said detection of dioxin in a food product comprises a maximum residue limits (MRL) of at most 10 pg/kg, preferably of at most 5 pg/kg, most preferably of at most 1 pg/kg.
The present invention will be further detailed in the following examples:
Examples
Clean-up of gelatin / protein containing matrix
Weigh 1.5 to 2.5 g of product into a 50 ml shaker tube, add 20 pi of 13-C labelled internal standard solution, 20 ml of 6M H2SO4 and shake vigorously. Leave the measuring cylinder in a water bath at 100 ° C for 1 hour (shaking occasionally). Allow to cool to room temperature and extract the resulting emulsion with 25 ml of hexane / iso-propanol mixture. Transfer the top layer (non-polar) quantitatively to a 25 ml graduated cylinder. Repeat the extraction with 25 ml hexane two more times, each time transferring the top layer to a round bottom flask. Evaporate the obtained extract to 1 -2 ml.

Claims (10)

ClausesClauses 1. Method for establishing the presence of dioxin in a food product, comprising the steps of1. Method for establishing the presence of dioxin in a food product, including the steps of a) contacting of a sample of the food product having a protein content of at least 15 wl%, based on the total weight of the sample, with a solution comprising sulphuric acid (H2SO4),a) contacting a sample of the food product with a protein content or at least 15 wl%, based on the total weight of the sample, with a solution including sulphuric acid (H 2 SO 4 ), b) heating of the sample contacted with the sulphuric acid solution of step a to at least 100 °C for at least 30 min, preferably 1 hour, to obtain an emulsion,b) heating the sample contacted with the sulphuric acid solution or step at least 100 ° C for at least 30 minutes, preferably 1 hour, to obtain an emulsion, c) allowing the emulsion to cool to a temperature between 15 and 25 °C,c) allowing the emulsion to cool to a temperature between 15 and 25 ° C, d) contacting the emulsion of step c with an extraction liquid comprised of at least 15 wt% hexane and at least 75 wt% isopropanol, based on the total weight of the extraction liquid,d) contacting the emulsion or step c with an extraction liquid comprised or at least 15 wt% hexane and at least 75 wt% isopropanol, based on the total weight of the extraction liquid, e) homogenizing of the emulsion of step d to form a suspension,e) homogenizing of the emulsion or step to form a suspension, f) allowing separation of the suspension in at least two phases and subsequently isolating the non-polar phase after extraction,f) allowing separation of the suspension in at least two phases and subsequently isolating the non-polar phase after extraction, g) contacting the isolated non-polar phase with hexane,g) contacting the isolated non-polar phase with hexane, h) repeating step f) and step g) for at least one more time, to obtain a sample A,h) repeating step f) and step g) for at least one more time, to obtain a sample A, j) subjecting sample A to a purification column comprised of AgNCh silica gel for purification of the sample A to obtain a sample B,j) subjecting sample A to a purification column comprised of AgNCh silica gel for purification or the sample A to obtain a sample B, k) subjecting the sample B to gas chromatography and/or mass spectrometry for establishing the presence of dioxin.k) subjecting the sample B to gas chromatography and / or mass spectrometry for establishing the presence of dioxin. 2. Method according to clause 1, wherein after step f), in addition to the non-polar phase, a polar phase is obtained and wherein said polar phase is subjected to a purification column comprised of H2SO4 silica gel and eluted with hexane to obtain a sample C.2. Method according to clause 1, after step f), in addition to the non-polar phase, a polar phase is obtained and said said polar phase is subject to a purification column comprised of H 2 SO 4 silica gel and eluted with hexane to obtain a sample C. 3. Method according to clause 2, wherein the polar phase is subjected to a purification column comprised of a glass fibre membrane to obtain sample C, followed by subjecting the sample C to gas chromatography and/or mass spectrometry for establishing the presence of dioxin.3. Method according to clause 2, the polar phase is subject to a purification column comprised of a glass fiber membrane to obtain sample C, followed by subjecting the sample C to gas chromatography and / or mass spectrometry for establishing the presence of dioxin. 4. Method according to any of the clauses 1 to 3, wherein in step a), the weight ratio between sample and sulphuric acid solution is at least 1:12, preferably at least 1:16, more preferably at least 1: 20, most preferably 1:24.4. Method according to any of the clauses 1 to 3, in step a), the weight ratio between sample and sulphuric acid solution is at least 1:12, preferably at least 1:16, more preferably at least 1:20, most preferably 1:24. 5. Method according to any of the clauses 1 to 4, wherein the sulphuric acid solution is at least a 5M solution, more preferably at least a 6M solution.5. Method according to any of the clauses 1 to 4, the sulphuric acid solution is at least a 5M solution, more preferably at least a 6M solution. 6. Method according to any of the clauses 1 to 5, wherein in step d the volume ratio of said extraction liquid in relation to said emulsion is at least 1:1, more preferably at least 5:1.6. Method according to any of the clauses 1 to 5, in step by volume ratio or said extraction liquid in relation to said emulsion is at least 1: 1, more preferably at least 5: 1. 7. Method according to any of the clauses 1 to 6, wherein in step g the volume ratio of hexane in relation to said non-polar phase is at least 1:1, more preferably at least 5:1.7. Method according to any of the clauses 1 to 6, in step g the volume ratio or hexane in relation to said non-polar phase is at least 1: 1, more preferably at least 5: 1. 8. Method according to any one of clauses 1 to 7, wherein said dioxin, furan and/or PCB is one or more selected from the group consisting of polychlorinated dibenzo-p-dioxins (PCDDs), derivatives of dibenzo-p-dioxin, 2,3,7,8-Tetrachlorodibenzodioxin (TCDD), polychlorinated dibenzofurans (PCDFs), furans and polychlorinated/polybrominated biphenyls (PCBs/PBBs).8. Method according to any one of clauses 1 to 7, said dioxin, furan and / or PCB is one or more selected from the group consisting of polychlorinated dibenzo-p-dioxins (PCDDs), derivatives or dibenzo-p-dioxin, 2,3,7,8-Tetrachlorodibenzodioxin (TCDD), polychlorinated dibenzofurans (PCDFs), furans and polychlorinated / polybrominated biphenyls (PCBs / PBBs). 9. Method according to any one of clauses 1 to 8, wherein said food product is one or more selected from the group consisting of meat and dairy products, fish, shellfish, fruit, vegetables, seeds, grain, herbs, spices, eggs and milk, preferably meal and dairy products, fish and shellfish.9. Method according to any one of clauses 1 to 8, said said food product is one or more selected from the group consisting of meat and dairy products, fish, shellfish, fruit, vegetables, seeds, grain, herbs, spices, eggs and milk, preferably meal and dairy products, fish and shellfish. 10. Method according to any one of clauses 1 lo 9, wherein said detection of dioxin in a food product comprises a maximum residue limits (MRL) of at most 10 pg/kg, preferably of at most 5 pg/kg, most preferably of at most 1 pg/kg.10. Method according to any one of clauses 1 to 9, said detection of dioxin in a food product comprises a maximum residue limit (MRL) or at most 10 pg / kg, preferably or at most 5 pg / kg, most preferably or at must 1 pg / kg. ConclusiesConclusions 1. Werkwijze voor het vaststellen van de aanwezigheid van dioxine in een voedingsproduct, omvattende de stappen vanA method for determining the presence of dioxin in a food product, comprising the steps of a) het in contact brengen van een monster van het voedingsproduct met een proteïnegehalte van ten minste 15 gew.%, op basis van het totale gewicht van het monster, met een zwavelzuur (H2SO4) bevattende oplossinga) contacting a sample of the food product with a protein content of at least 15% by weight, based on the total weight of the sample, with a solution containing sulfuric acid (H 2 SO 4 ) b) het verwarmen van hel monster dat in contact is geweest met de zwavelzuuroplossing van stap a tol ten minste 100 ° C, gedurende ten minste 30 minuten, bij voorkeur 1 uur, om een emulsie te verkrijgen,b) heating the sample that has been in contact with the sulfuric acid solution of step a at least 100 ° C for at least 30 minutes, preferably 1 hour, to obtain an emulsion, c) de emulsie laten afkoelen tot een temperatuur tussen 15 en 25 °C,c) allowing the emulsion to cool to a temperature between 15 and 25 ° C, d) het in contact brengen van de emulsie van stap c met een extractievloeistof bestaande uit ten minste 15 gew.% hexaan en ten minste 75 gew.% isopropanol, gebaseerd op het totale gewicht van de extractievloeistof,d) contacting the emulsion of step c with an extraction liquid consisting of at least 15% by weight of hexane and at least 75% by weight of isopropanol, based on the total weight of the extraction liquid, e) het homogeniseren van de emulsie van stap d om een suspensie te vormen,e) homogenizing the emulsion of step d to form a suspension, f) het laten scheiden van de suspensie in ten minste twee fasen en vervolgens het isoleren van de apolaire fase na extractie,f) separating the suspension into at least two phases and then isolating the non-polar phase after extraction, g) het in contact brengen van de geïsoleerde apolaire fase met hexaan,g) contacting the isolated apolar phase with hexane, h) herhalen van stap f) en stap g) voor ten minste nog één keer, om een monster A te verkrijgen,h) repeating step f) and step g) for at least one more time to obtain a sample A, j) het onderwerpen van monster A aan een zuiveringskolom bestaande uit AgN03 silicagel voor zuivering van het monster A om een monster B te verkrijgen,j) subjecting sample A to a purification column consisting of AgNO 3 silica gel for purification of sample A to obtain sample B, k) het onderwerpen van monster B aan gaschromatografie en / of massaspectrometrie voor het vaststellen van de aanwezigheid van dioxine.k) subjecting sample B to gas chromatography and / or mass spectrometry to determine the presence of dioxin. 2. Werkwijze volgens conclusie 1, waarbij na stap f), in aanvulling op de apolaire fase, een polaire fase wordt verkregen en waarbij de bodemfase wordt onderworpen aan een zuiveringskolom bestaande uil H2SO4 silicagel en wordt geëlueerd met hexaan om een monster te verkrijgen C.A method according to claim 1, wherein after step f), in addition to the apolar phase, a polar phase is obtained and wherein the bottom phase is subjected to a purification column consisting of owl H 2 SO 4 silica gel and eluted with hexane to obtain a sample C. 3. Werkwijze volgens conclusie 2, waarbij de polaire fase wordt onderworpen aan een zuiveringskolom bestaande uit een glasvezelmembraan om monster C te verkrijgen, gevolgd door het onderwerpen van monster C aan gaschromatografie en / of massaspectrometrie voor het vaststellen van de aanwezigheid van dioxine.The method of claim 2, wherein the polar phase is subjected to a purification column consisting of a glass fiber membrane to obtain sample C, followed by subjecting sample C to gas chromatography and / or mass spectrometry to determine the presence of dioxin. 4. Werkwijze volgens één van de conclusies 1 tot en met 3, waarbij in stap a) de gewichtsverhouding tussen monster en zwavelzuuroplossing ten minste 1:12 is, bij voorkeur ten minste 1:16, met meer voorkeur ten minste 1:20, met de meeste voorkeur 1:24.A method according to any of claims 1 to 3, wherein in step a) the weight ratio between sample and sulfuric acid solution is at least 1:12, preferably at least 1:16, more preferably at least 1:20, with most preferred 1:24. 5. Werkwijze volgens één van de conclusies 1 tot en met 4, waarbij de zwavelzuuroplossing ten minste een 5M oplossing, met meer voorkeur ten minste een 6M oplossing is.The method of any one of claims 1 to 4, wherein the sulfuric acid solution is at least a 5M solution, more preferably at least a 6M solution. 6. Werkwijze volgens één van de conclusies 1 tot en met 5, waarbij in stap d de vol urne verhouding van de genoemde extractievloeistof ten opzichte van de genoemde emulsie ten minste 1:1, met meer voorkeur ten minste 5: 1 is.A method according to any one of claims 1 to 5, wherein in step d, the full-volume ratio of said extraction liquid to said emulsion is at least 1: 1, more preferably at least 5: 1. 7. Werkwijze volgens één van de conclusies 1 tot en met 6, waarbij in stap g de volumeverhouding van hexaan ten opzichte van de apolaire fase ten minste 1:1, met meer voorkeur ten minste 5:1 is.The method according to any of claims 1 to 6, wherein in step g the volume ratio of hexane to the non-polar phase is at least 1: 1, more preferably at least 5: 1. 8. Werkwijze volgens één van de conclusies 1 lot en met 7, waarbij het dioxine, furan en / of PCB één of meer is gekozen uit de groep bestaande uit polychloordibenzo-p-dioxinen (PCDD's), derivaten van dibenzo-p-dioxine, 2,3,7,8-Tetraehlorodibenzodioxin (TCDD), polychloordibenzofuranen (PCDF's), furanen en polygechloreerde / polygebromeerde bifenylen (PCB's / PBB's).A method according to any of claims 1 to 7, wherein the dioxin, furan and / or PCB is one or more selected from the group consisting of polychlorodibenzo-p-dioxins (PCDDs), derivatives of dibenzo-p-dioxin, 2,3,7,8-tetrachlorodibenzodioxin (TCDD), polychlorinated dibenzofurans (PCDFs), furans and polychlorinated / polybrominated biphenyls (PCBs / PBBs). 9. Werkwijze volgens één van de conclusies 1 tot en met 8, waarbij het voedingsproduct éé of meer is gekozen uit de groep bestaande uit vlees en zuivelproducten, vis, schaaldieren, fruit, groenten, zaden, graan, kruiden, specerijen, eieren en melk, bij voorkeur vlees en zuivelproducten, vis en schaaldieren.A method according to any of claims 1 to 8, wherein the food product is one or more selected from the group consisting of meat and dairy products, fish, shellfish, fruit, vegetables, seeds, grain, herbs, spices, eggs and milk , preferably meat and dairy products, fish and shellfish. 10. Werkwijze volgens één van de conclusies 1 tot en met 9, waarbij de detectie van dioxine in een voedingsproduct een maximum residulimiet (MRL) omvat van ten hoogste 10 pg / kg, bij voorkeur van ten hoogste 5 pg / kg, met de meeste voorkeur van maximaal 1 pg / kg.The method of any one of claims 1 to 9, wherein the detection of dioxin in a food product comprises a maximum residue limit (MRL) of at most 10 pg / kg, preferably of at most 5 pg / kg, with most preferably of a maximum of 1 pg / kg.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332819A1 (en) * 1988-01-26 1989-09-20 The Regents Of The University Of California Method for the immunoassay of dioxins and dibenzofurans
RU2188418C1 (en) * 2000-12-28 2002-08-27 Научно-исследовательский институт проблем хранения Method of assay of fats and fat-containing foodstuffs
JP2009097886A (en) * 2007-10-12 2009-05-07 Chubu Electric Power Co Inc Simplified measuring method of dioxins
US8507292B2 (en) * 2008-02-28 2013-08-13 Dionex Corporation Sample pretreatment and extraction

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0332819A1 (en) * 1988-01-26 1989-09-20 The Regents Of The University Of California Method for the immunoassay of dioxins and dibenzofurans
RU2188418C1 (en) * 2000-12-28 2002-08-27 Научно-исследовательский институт проблем хранения Method of assay of fats and fat-containing foodstuffs
JP2009097886A (en) * 2007-10-12 2009-05-07 Chubu Electric Power Co Inc Simplified measuring method of dioxins
US8507292B2 (en) * 2008-02-28 2013-08-13 Dionex Corporation Sample pretreatment and extraction

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