EN 16995:2017

Foodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) with on-line HPLC-GC-FID analysis

This European Standard specifies a highly efficient method for the determination of saturated and aromatic hydrocarbons (from C10 to C50) in vegetable fats and oils and foodstuff on basis of vegetable oils for which it has been interlaboratory validated, with online-HPLC-GC-FID [1], [2] and [3]. This standard is not intended to be applied to other matrices.
The method can be used for the analysis of mineral oil saturated hydrocarbons (MOSH) and/or mineral oil aromatic hydrocarbons (MOAH).
The method has been tested in an interlaboratory study via the analysis of both naturally contaminated and spiked vegetable oil samples and mayonnaise and margarine samples, ranging from 4 mg/kg to 197 mg/kg for MOSH, and from 2 mg/kg to 51 mg/kg for MOAH.
According to the results of the interlaboratory studies, the method has been proven suitable for MOSH- and MOAH mass concentrations each above 10 mg/kg.
In case of suspected interferences from natural sources, the mineral origin of the MOSH and MOAH fraction can be verified by examination of the pattern by GC-MS.

Lebensmittel - Pflanzliche Öle und Lebensmittel auf Basis pflanzlicher Öle - Bestimmung von gesättigten Mineralöl-Kohlenwasserstoffen (MOSH) und aromatischen Mineralöl-Kohlenwasserstoffen (MOAH) mit on-line HPLC-GC-FID

Diese Europäische Norm legt ein hocheffizientes Verfahren zur Bestimmung von gesättigten und aromatischen Kohlenwasserstoffen (von C10 bis C50) in pflanzlichen Fetten und Ölen und in Lebensmitteln auf der Basis von Pflanzenölen, die im Ringversuch getestet wurden, fest. Das Verfahren nutzt die Online-HPLC-GC-FID [1], [2], [3] (HPLC: Hochleistungsflüssigchromatographie, en: high performance liquid chromatography; GC: Gaschromatographie, en: gas chromatographie; FID: Flammenionisationsdetektor, en: flame ionization detector). Diese Norm ist nicht dafür vorgesehen, auf andere Lebensmittel angewendet zu werden, als die, die im Ringversuch getestet wurden.
Das Verfahren kann zur Bestimmung von gesättigten Mineralöl-Kohlenwasserstoffen (MOSH) und/oder aromatischen Mineralöl-Kohlenwasserstoffen (MOAH) verwendet werden.
Das Verfahren wurde in einem Ringversuch sowohl an natürlich kontaminierten als auch an angereicherten Pflanzenölproben, Mayonnaise und Margarine in Bereichen von 4 mg/kg bis 197 mg/kg für MOSH und von 2 mg/kg bis 51 mg/kg für MOAH getestet.
Nach den Ringversuchsergebnissen hat sich das Verfahren bei Massenkonzentrationen an MOSH und MOAH von jeweils über 10 mg/kg als geeignet erwiesen.
Wenn Störungen aus natürlichen Quellen vermutet werden, kann die mineralische Herkunft des MOSH- und MOAH-Anteiles durch Untersuchung des chromatographischen Profils durch GC-MS verifiziert werden.

Produits alimentaires - Huiles végétales et produits alimentaires à base d'huiles végétales - Dosage des hydrocarbures saturés d’huile minérale (MOSH) et des hydrocarbures aromatiques d’huile minérale (MOAH) par analyse par CLHP-CG-FID en ligne

La présente Norme européenne spécifie une méthode hautement efficace applicable pour le dosage des hydrocarbures saturés et aromatiques (de C10 à C50) présents dans les corps gras végétaux et les produits alimentaires à base d’huiles végétales, pour lesquels elle a fait l’objet d’une validation interlaboratoires, par CLHP-CG-FID en ligne [1], [2] et [3]. La présente norme n’est pas destinée à être appliquée à d’autres matrices.
La méthode peut être employée pour l’analyse des hydrocarbures saturés d’huile minérale (MOSH) et/ou des hydrocarbures aromatiques d’huile minérale (MOAH).
Cette méthode a été soumise à essai lors d’une étude interlaboratoires en procédant à l’analyse d’échantillons d’huile végétale naturellement contaminés et dopés et d’échantillons de mayonnaise et de margarine à des teneurs comprises entre 4 mg/kg et 197 mg/kg pour les MOSH et entre 2 mg/kg et 51 mg/kg pour les MOAH.
Conformément aux résultats des études interlaboratoires, il a été démontré que la méthode est adaptée pour les concentrations massiques de MOSH et de MOAH au-dessus de 10 mg/kg chacune.
En cas de suspicion d’interférences dues à des sources naturelles, l’origine minérale de la fraction de MOSH et de MOAH peut être vérifiée par un examen du profil par CG-SM.

Živila - Rastlinska olja in živila na osnovi rastlinskih olj - Določevanje mineralnih olj nasičenih ogljikovodikov (MOSH) in mineralnih olj aromatskih ogljikovodikov (MOAH) z analizo on-line HPLC-GC-FID

Ta evropski standard opredeljuje metodo za določanje nasičenih in aromatskih ogljikovodikov (od C10 do C50) v rastlinskih maščobah, oljih in živilih na osnovi rastlinskih olj z analizo on-line HPLC-GC-FID [1], [2]. Analiza HPLC-GC-FID je zelo učinkovita metoda za določanje mineralnih olj v različnih živilih [3].
To metodo je mogoče uporabiti za analizo mineralnih olj nasičenih ogljikovodikov (MOSH) in/ali mineralnih olj aromatskih ogljikovodikov (MOAH). Mineralna olja nasičenih ogljikovodikov (MOSH) so parafinski (odprtoverižni, običajno razvejani) in naftenski (ciklični, alkilirani) ogljikovodiki; mineralna olja aromatskih ogljikovodikov (MOAH) so aromatski in v večji meri alkilirani ogljikovodiki.
Ta metoda je bila preskušena v medlaboratorijski študiji z analizo naravno kontaminiranih in primešanih vzorcev rastlinskih olj, majoneze ter margarine v razponu 4–197 mg/kg za mineralna olja nasičenih ogljikovodikov in 2–51 mg/kg za mineralna olja aromatskih ogljikovodikov.
Ta metoda je dokazano primerna za vsebnosti nad 10 mg/kg na podlagi rezultatov medlaboratorijskih preskusov.

General Information

• Status: Withdrawn
• Publication Date: 13-Jun-2017
• Withdrawal Date: 13-Apr-2025
• ICS: 67.200.10 - Animal and vegetable fats and oils
• Technical Committee: CEN/TC 275 - Food analysis - Horizontal methods
• Drafting Committee: CEN/TC 275/WG 13 - Neoformed contaminants
• Current Stage: 9960 - Withdrawal effective - Withdrawal
• Start Date: 17-Jan-2024
• Completion Date: 14-Apr-2025
• Directive: 882/2004 - Regulation (EC) No 882/2004 of the European Parliament and of the Council of 29 April 2004 onofficial controls performed to ensure the verification of compliance with feed and food law, animal health and animal welfare rules
• Mandate: M/463 - Mandate for Standardisation addressed to CEN for the methods of analysis for food contaminants
• Ref Project: SIST EN 16995:2017 - Foodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) with on-line HPLC-GC-FID analysis

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.RJOMLNRYRGLNRYLebensmittel - Pflanzliche Öle und Lebensmittel auf Basis pflanzlicher Öle - Bestimmung von Mineralölen aus gesättigten Kohlenwasserstoffen (MOSH) und aus aromatischen Kohlenwasserstoffen (MOAH) mit on-line HPLC-GC-FIDProduits alimentaires - Huiles végétales et denrées à base d'huiles végétales - Dosage des hydrocarbures saturés d’huile minérale (MOSH) et des hydrocarbures aromatiques d’huile minérale (MOAH) par analyse par CLHP-CG-FID en ligneFoodstuffs - Vegetable oils and foodstuff on basis of vegetable oils - Determination of mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH) with on-line HPLC-GC-FID analysis67.200.10Animal and vegetable fats and oilsICS:Ta slovenski standard je istoveten z:EN 16995:2017SIST EN 16995:2017en,fr,de01-september-2017SIST EN 16995:2017SLOVENSKI
STANDARD
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN 16995
June
t r s y ICS
x yä t r rä s r English Version
Foodstuffs æ Vegetable oils and foodstuff on basis of vegetable oils æ Determination of mineral oil saturated Produits alimentaires æ Huiles végétales et produits alimentaires à base d 5huiles végétales æ Dosage des par analyse par CLHPæCGæFID en ligne
Lebensmittel æ Pflanzliche Öle und Lebensmittel auf Basis pflanzlicher Öle æ Bestimmung von gesättigten mit onæline HPLCæGCæFID This European Standard was approved by CEN on
s r March
t r s yä
egulations which stipulate the conditions for giving this European Standard the status of a national standard without any alterationä Upætoædate lists and bibliographical references concerning such national standards may be obtained on application to the CENæCENELEC Management Centre or to any CEN memberä
translation under the responsibility of a CEN member into its own language and notified to the CENæCENELEC Management Centre has the same status as the official versionsä
CEN members are the national standards bodies of Austriaá Belgiumá Bulgariaá Croatiaá Cyprusá Czech Republicá Denmarká Estoniaá Finlandá Former Yugoslav Republic of Macedoniaá Franceá Germanyá Greeceá Hungaryá Icelandá Irelandá Italyá Latviaá Lithuaniaá Luxembourgá Maltaá Netherlandsá Norwayá Polandá Portugalá Romaniaá Serbiaá Slovakiaá Sloveniaá Spainá Swedená Switzerlandá Turkey and United Kingdomä
EUROPEAN COMMITTEE FOR STANDARDIZATION COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre:
Avenue Marnix 17,
B-1000 Brussels
t r s y CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Membersä Refä Noä EN
s x { { wã t r s y ESIST EN 16995:2017

Page European foreword . 3 1 Scope . 4 2 Normative references . 4 3 Terms and definitions . 4 4 Principle . 4 5 Reagents . 5 6 Apparatus . 8 7 Sample storage . 9 8 Preparation of the test sample . 10 9 Preparation of the analytical sample . 10 10 Liquid chromatography and gas chromatography . 12 11 Precision . 17 12 Test report . 17 Annex A (informative)
Examples of chromatograms . 18 Annex B (informative)
Precision data . 32 Bibliography . 35
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{ { %. 5.5 n-Hexane, trace organic analysis grade, for pesticide residue analysis. n-Hexane purity can be checked by concentrating 30 ml of n-hexane mixed with 25 µl of internal standard solution (5.21) and 2 drops of bis(2-ethylhexyl) maleate (5.29) using a rotary evaporator, dissolving the residue in 0,2 ml of n-hexane and the analysis of 50 µl by online-HPLC-GC-FID (6.8). Take care that in the evaporation step the residue is not evaporated to dryness to avoid loss of volatile hydrocarbons. The signal abundance of the residue after evaporation should not exceed a tenth of the signal abundance obtained at the quantification limit.
1) Silica gel is available from Merck, reference 7754 or 7734 (www.merck-chemicals.com). It is an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 16995:2017

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{ {
¨ä DCM purity can be checked by concentrating 50 ml of DCM mixed with 25 µl of internal standard solution (5.21) and 2 drops of bis(2-ethylhexyl) maleate (5.29) using a rotary evaporator, dissolving the residue in 0,2 ml of n-hexane and the analysis of 50 µl by online-HPLC-GC-FID (6.8). Take care that in the evaporation step the residue is not evaporated to dryness to avoid loss of volatile hydrocarbons. The signal abundance of the residue after evaporation should not exceed a fifth of the signal abundance obtained at the quantification limit. 5.7 Dichloromethane solution. Mix 30 ml DCM (5.6) with n-hexane (5.5) up to a volume of 100 ml. The solution should be freshly prepared daily. 5.8 Toluene. 5.9 1,1,2-Trichloroethane. 5.10 Perylene (Per %. 5.11 5-ƒ-Cholestane (Cho), purity
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{ y %. 5.12 n-Undecane (n-C11), purity
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{ z %. 5.13 n-Tridecane (n-C13), purity
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{ y %. 5.14 Tri-tert-butylbenzene (TBB). 5.15 Bicyclohexyl (CyCy)á purity
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{ { %. 5.16 1-Methylnaphthalene (1-MN)á purity
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{ w %. 5.17 2-Methylnaphthalene (2-MN)á purity
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{ y %. 5.18 Pentylbenzene (PB)á purity
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{6 %. 5.19 Stock solutions, mass concentration
= 10 mg/ml. Prepare individual stock solutions by weighing, to the nearest 1 mg, 100 mg of n-C11 (5.12), n-C13 (5.13), TBB (5.14), CyCy (5.15), 1-MN (5.16), 2-MN (5.17) and PB (5.18) into a 10 ml volumetric flask and dilute to the mark with 1,1,2-trichloroethane (5.9) or toluene (5.8). Store the solutions at room temperature. If crystals precipitate during storage, warm the solution until everything has dissolved. 5.20 Internal standard solution 1 (ISTD1)2). Weigh, to the nearest 0,5 mg, 12 mg of Per (5.10) and Cho (5.11) in a volumetric flask of 20 ml (6.22), to which 600 µl of each stock solution (5.19) is added with the exception of n-C13, of which 300 µl is added. Fill the volumetric flask up to 20 ml with 1,1,2-trichloroethane (5.9) or toluene (5.8). Resulting mass concentrations are for n-C13:
= 150 µg/ml, for n-C11, TBB, CyCy, 1-MN, 2-MN and PB:
= 300 µg/ml and for Per, Cho:
= 600 µg/ml.
2) This standard mixture is available by e.g. Restek Corp., Cat.# 31070. It is an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 16995:2017

= 5 µg/ml, for n-C11, TBB, CyCy, 1-MN, 2-MN and PB:
= 10 µg/ml and for Per, Cho:
= 20 µg/ml. 5.22 Aluminium oxide 90, alkaline, for column chromatography 0,063 mm to 0,2 mm. 5.23 Aluminium oxide, activated (ALOX). Condition aluminium oxide 90 (5.22) for at least 16 h at 500 °C in an oven before using. 5.24 Chloroperbenzoic acid (CPBA), purity 70 % to 75 %. 5.25 CPBA solution,
= 0,1 g/ml in dichloromethane. For example 1 g of CPBA (5.24) in 10 ml of DCM (5.6). Clouding of solution does not disturb the reaction. The solution can be used for up to one week. 5.26 Ascorbic acid. 5.27 Silica-ALOX column. Insert a filter (6.3) in each glass column (6.2). Then, fill in 10 g of ALOX (5.23) and 3 g of silica gel (5.2) and compress. 5.28 Cleanup column. Insert a filter (6.3) in a glass SPE tube (6.20). Then, fill in 3 g of silica gel (5.2), compress and overlay with 0,5 g of sodium sulfate (5.4). 5.29 Keeper solvent. The keeper is a solvent that will not evaporate or evaporate to a lesser degree during the evaporation step, e.g. bis(2-ethylhexyl) maleate. A keeper is used to enhance the recovery of volatile compounds. 5.30 Carrier gas for gas chromatographyá preferably hydrogená purity
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{ {á { { w %. 5.31 Auxiliary gases for flame ionization detector, hydrogen, air, and nitrogen suitable for gas chromatography. 5.32 Alkane standard mixture C10 to C40, solution of equal concentration in an apolar solvent,
= 1 µg/ml. 5.33 Ethanol, absolute. NOTE The ethanol purity can be checked by concentrating 50 ml of ethanol mixed with 25 µl of internal standard solution (5.21) using a rotary evaporator, dissolving the residue in 0,2 ml of n-hexane and the analysis of 50 µl by online-HPLC-GC-FID (6.8). 5.34 Mixture of ethanol and n-hexane %. Mix 50 ml of ethanol (5.33) with 50 ml of n-hexane (5.5). 5.35 n-Pentacontane (n-C50), purity
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{ z %. SIST EN 16995:2017
³ 10 µg/ml. Weigh 2 mg of C50 (5.35) in a volumetric flask of 20 ml (6.22) and dilute to the mark with toluene (5.8). Proceed to a second dilution of 1 ml in a 10 ml volumetric flask (6.22). Store the solutions at room temperature. NOTE 1 Solubility of pentacontane in toluene is limited at room temperature. However, the concentration of the solution of pentacontane does not need to be accurate as it is used only to determine the limit of integration for mineral oil peak. NOTE 2 It is also possible to use a commercial mixture of n-alkanes from C12 to C60 that contains n-pentacontane3). 5.37 Sodium carbonate solution,
= 0,1 g/ml in water (5.3). 5.38 Mixture of DCM and n-hexane. Mix 20 ml DCM (5.6) with n-hexane (5.5) up to a volume of 100 ml. The solution should be freshly prepared daily. 5.39 Blank refined sunflower oil. 6 Apparatus Usual laboratory apparatus and, in particular, the following. The glassware shall be thoroughly cleaned and rinsed with n-hexane (5.5) or baked in an oven before use so that it is free from impurities. 6.1 Centrifuge and centrifuge tubes. 6.2 Glass column for cleanup, 15 cm to 20 cm length and 15 mm to 20 mm internal diameter. 6.3 Filter for glass column. 6.4 Glass vials with screw caps, volume of 40 ml. 6.5 Rotary evaporator, with vacuum and a water bath at 35 °C (recommended). Care should be taken to prevent cross contamination. Clean the system thoroughly between determinations. 6.6 Automatic evaporator.4) 6.7 Glass sample vials, volume of 2 ml. 6.8 High performance liquid chromatograph, coupled with gas chromatograph and flame ionization detector (HPLC-GC-FID).
3) ASTM D5442 C12-C60 Qualitative Retention Time Mix is available by e.g. Supelco Cat.# 500623. It is an example of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. 4) For example, MicroDancer, IR-Dancer (e.g. Zinser) or Syncore Analyst (Büchi). These are examples of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 16995:2017

5) These are examples of a suitable product available commercially. This information is given for the convenience of users of this European Standard and does not constitute an endorsement by CEN of this product. Equivalent products may be used if they can be shown to lead to the same results. SIST EN 16995:2017

9.3 and/or 9.4. The amount of the added internal standards may be increased in order to lower the impact of the matrix interferences, if necessary. 9.3 Cleanup for MOSH fraction with aluminium oxide (optional) Weigh, to the nearest 1 mg, 300 mg of sample in a 40 ml glass vial (6.4). Add 30 µl of ISTD2 (5.21) and dissolve the mixture in 2 ml of n-hexane (5.5). Subsequently, transfer the sample solution to a silica-ALOX column (5.27) that was rinsed by using 20 ml of n-hexane prior to sample loading. Elute the aliphatic hydrocarbon fraction with 25 ml of n-hexane. Evaporate the solvent under reduced pressure after addition of 2 drops of bis(2-ethylhexyl) maleate (5.29) at 40 °C. Take care that in the evaporation step the residue is not evaporated to dryness to avoid loss of volatile hydrocarbons. Dissolve the
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