EN 1186-2:2002

Materials and articles in contact with foodstuffs - Plastics - Part 2: Test methods for overall migration into olive oil by total immersion

This part of this European Prestandard describes test methods for the determination of the overall migration from plastics intended to come into contact with a fatty foodstuff, by total immersion in olive oil for 10 days, 24 h or for 2 h at 40 °C or 2 h at 70 °C. This method is most suitable for plastics in the form of films and sheets, but can be applied to a wide range of articles or containers from which test pieces of a suitable size can be cut. The test method described is applicable to most types of plastics, althoug there are some plastics for which it is known not to be applicable.

Werkstoffe und Gegenstände in Kontakt mit Lebensmitteln - Kunststoffe - Teil 2: Prüfverfahren für die Gesamtmigration in Olivenöl durch völliges Eintauchen

1 Anwendungsbereich
Dieser Teil der Europäischen Norm beschreibt Prüfverfahren zur Bestimmung der Gesamtmigration aus Werkstoffe und Gegenständen aus Kunststoff in Simulanzlösemittel für fetthaltige Lebensmittel durch völliges Eintauchen der Prüfmuster in das Simulanzlösemittel bei Temperaturen über 20°C bis zu (jedoch nicht einschließlich) 100°C während bestimmter Zeiten.
Dieses Verfahren ist bestens geeignet für Kunststoffe in Form von Folien und Platten, kann aber auch angewendet werden für einen weiten Bereich von Gegenständen und Behältern, aus denen Prüfabschnitte geeigneter Größe geschnitten werden können.
ANMERKUNG   Dieses Prüfverfahren wurde beschrieben für die Verwendung von Olivenöl als Simulanzlösemittel für fetthaltige Lebensmittel. Das Prüfverfahren kann mit entsprechender Modifikation auch mit ¿anderen Simulanzlösemitteln für fetthaltige Lebensmittel" angewendet werden, dem sogenannten Simulanzlösemittel D - einer synthetischen Mischung von Triglyceriden, Sonnenblumenöl und Maiskeimöl. Diese anderen Simulanzlösemittel für fetthaltige Lebensmittel ergeben für ihre Methylester andere Chromatogramme als für die Methylester des Simulanzlösemittels Olivenöl. Aus den Chromatogrammen der Methylester der anderen Simulanzlösemittel werden geeignete Peaks zur quantitativen Bestimmung des aus dem Prüfmuster extrahierten Simulanzlösemittels ausgewählt.
Das beschriebene Prüfverfahren kann auf die meisten Kunststoffsorten angewendet werden. Es gibt jedoch einige Kunststoffe, von denen bekannt ist, dass es nicht angewendet werden kann.

Matériaux et objets en contact avec les denrées alimentaires - Matière plastique - Partie 2: Méthodes d'essai pour la migration globale dans l'huile d'olive par immersion totale

La présente partie de cette norme européenne décrit une méthode d'essai de vérification de la migration globale des matériaux et objets en matière plastique destinés à entrer en contact avec une denrée alimentaire grasse, par immersion totale d'éprouvettes dans un simulant gras à des températures supérieures à 20 °C mais inférieures à 100 °C pendant des durées définies.
Cette méthode est plus particulièrement applicable aux matières plastiques sous forme de films et de feuilles, mais peut être aussi appliquée à une large gamme d'objets ou de récipients à partir desquels des morceaux de taille appropriée peuvent être prélevés.
NOTE   La présente méthode d'essai a été écrite pour l'huile d'olive. Elle peut également être utilisée, avec les modifications qui s'imposent, pour d'autres simulants gras appelés du terme générique « simulant D », mélange synthétique de triglycérides, d'huile de tournesol et d'huile de maïs. Ces autres simulants gras donneront des chromatogrammes des esters méthyliques du simulant utilisé différents de ceux des esters méthyliques de l'huile d'olive. Choisir pour le dosage quantitatif du simulant extrait des éprouvettes les pics de chromatogramme correspondant aux esters méthyliques des autres simulants gras.
La méthode d'essai décrite est applicable à la plupart des types de matières plastiques, bien qu'il soit connu qu'elle ne le soit pas à certains.

Materiali in predmeti v stiku z živili - Polimerni materiali - 2. del: Preskusne metode za celotno migracijo v olivno olje s popolno potopitvijo

General Information

• Status: Withdrawn
• Publication Date: 16-Apr-2002
• Withdrawal Date: 13-Apr-2025
• ICS: 67.250 - Materials and articles in contact with foodstuffs
• Technical Committee: CEN/TC 194 - Utensils in contact with food
• Drafting Committee: CEN/TC 194/SC 1/WG 1 - Overall migration
• Current Stage: 9960 - Withdrawal effective - Withdrawal
• Start Date: 27-Jul-2022
• Completion Date: 14-Apr-2025
• Directive: 89/109/EEC - Materials and articles intended to come into contact with foodstuffs
• Mandate: M/052 - Standardization mandate to CEN concerning methods of analysis for materials and articles intended to come into contact with foodstuffs
• Ref Project: SIST EN 1186-2:2002 - Materials and articles in contact with foodstuffs - Plastics - Part 2: Test methods for overall migration into olive oil by total immersion

Relations

• Replaces: ENV 1186-2:1994 - Materials and articles in contact with foodstuffs - Plastics - Part 2: Test methods for overall migration into olive oil by total immersion
• Effective Date: 22-Dec-2008
• Replaced By: EN 1186-2:2022 - Materials and articles in contact with foodstuffs - Plastics - Part 2: Test methods for overall migration in vegetable oils
• Effective Date: 03-Aug-2022
• Revised: EN 1186-3:2022 - Materials and articles in contact with foodstuffs - Plastics - Part 3: Test methods for overall migration in evaporable simulants
• Effective Date: 24-Jan-2018

Standards Content (Sample)

2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.Materiali in predmeti v stiku z živili - Polimerni materiali - 2. del: Preskusne metode za celotno migracijo v olivno olje s popolno potopitvijoWerkstoffe und Gegenstände in Kontakt mit Lebensmitteln - Kunststoffe - Teil 2: Prüfverfahren für die Gesamtmigration in Olivenöl durch völliges EintauchenMatériaux et objets en contact avec les denrées alimentaires - Matiere plastique - Partie 2: Méthodes d'essai pour la migration globale dans l'huile d'olive par immersion totaleMaterials and articles in contact with foodstuffs - Plastics - Part 2: Test methods for overall migration into olive oil by total immersion67.250Materiali in predmeti v stiku z živiliMaterials and articles in contact with foodstuffsICS:Ta slovenski standard je istoveten z:EN 1186-2:2002SIST EN 1186-2:2002en01-september-2002SIST EN 1186-2:2002SLOVENSKI
STANDARDSIST ENV 1186-2:19971DGRPHãþD

EUROPEAN STANDARDNORME EUROPÉENNEEUROPÄISCHE NORMEN 1186-2April 2002ICS 67.250Supersedes ENV 1186-2:1994English versionMaterials and articles in contact with foodstuffs - Plastics - Part2: Test methods for overall migration into olive oil by totalimmersionMatériaux et objets en contact avec les denréesalimentaires - Matière plastique - Partie 2: Méthodesd'essai pour la migration globale dans l'huile d'olive parimmersion totaleWerkstoffe und Gegenstände in Kontakt mit Lebensmitteln- Kunststoffe - Teil 2: Prüfverfahren für dieGesamtmigration in Olivenöl durch völliges EintauchenThis European Standard was approved by CEN on 4 January 2002.CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this EuropeanStandard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such nationalstandards may be obtained on application to the Management Centre or to any CEN member.This European Standard exists in three official versions (English, French, German). A version in any other language made by translationunder the responsibility of a CEN member into its own language and notified to the Management Centre has the same status as the officialversions.CEN members are the national standards bodies of Austria, Belgium, Czech Republic, Denmark, Finland, France, Germany, Greece,Iceland, Ireland, Italy, Luxembourg, Malta, Netherlands, Norway, Portugal, Spain, Sweden, Switzerland and United Kingdom.EUROPEAN COMMITTEE FOR STANDARDIZATIONCOMITÉ EUROPÉEN DE NORMALISATIONEUROPÄISCHES KOMITEE FÜR NORMUNGManagement Centre: rue de Stassart, 36
B-1050 Brussels© 2002 CENAll rights of exploitation in any form and by any means reservedworldwide for CEN national Members.Ref. No. EN 1186-2:2002 ESIST EN 1186-2:2002

Determination of the suitability of olive oil as the fatty food simulant and oftriheptadecanoin as the internal standard.19Annex B (normative)
Determination of the need for sample conditioning.21Annex C (normative)
Determination of the need for sample conditioning and determination of themass of moisture sensitive test specimens, by vacuum drying.22Annex D (normative)
Determination of change in moisture content of test specimens bymeasurement of the transfer of water to, or from olive oil, by Karl Fischer titration.24Annex E (informative)
Typical chromatograms and calibration graph.26Annex F (informative)
Precision data.29Annex ZA (informative)
Relationship of this European Standard with Council Directive 89/109/EECand Commission Directive 90/128/EEC and associated Directives.30Bibliography.32SIST EN 1186-2:2002

Further Directives andamendments to existing Directives are expected which could change the legislative requirements which thisstandard supports.
It is therefore strongly recommended that users of this standard refer to the latest relevantpublished Directive(s) before commencement of any of the test or tests described in this standard.EN 1186-2 should be read in conjunction with EN 1186-1.Further Parts of this standard have been prepared concerned with the determination of overall migration fromplastics materials into food simulants.Their titles are as follows:EN 1186 Materials and articles in contact with foodstuffs - Plastics –Part 1Guide to the selection of conditions and test methods for overall migrationPart 3Test methods for overall migration into aqueous food simulants by total immersionPart 4Test methods for overall migration into olive oil by cellPart 5Test methods for overall migration into aqueous food simulants by cellPart 6Test methods for overall migration into olive oil using a pouchPart 7Test methods for overall migration into aqueous food simulants using a pouchPart 8Test methods for overall migration into olive oil by article fillingPart 9Test methods for overall migration into aqueous food simulants by article fillingPart 10Test methods for overall migration into olive oil (modified method for use in cases whereincomplete extraction of olive oil occurs)Part 11Test methods for overall migration into mixtures of
14C-labelled synthetic triglyceridePart 12Test methods for overall migration at low temperaturesSIST EN 1186-2:2002

The test method can also be usedwith appropriate modifications with 'other fatty food simulants ' called simulant D - a synthetic mixture of triglycerides, sunfloweroil and corn oil.
These other fatty food simulants will produce different chromatograms for the simulant methyl esters to those ofthe methyl esters of olive oil.
Select suitable chromatogram peaks of the methyl esters of the other fatty food simulants for thequantitative determination of the simulant extracted from the test specimens.The test method described is applicable to most types of plastics, although there are some plastics for which it isknown not to be applicable.2 Normative referencesThis European Standard incorporates by dated and undated reference, provisions from other publications.
Thesenormative references are cited at the appropriate places in the text, and the publications are listed hereafter. Fordated references, subsequent amendments to and revisions of any of these publications apply to this EuropeanStandard only when incorporated in it by amendment or revision.
For undated references the latest edition of thepublication referred to applies (including amendments).EN 1186-1:2002, Materials and articles in contact with foodstuffs - Plastics – Part 1:Guide to the selection ofconditions and test methods for overall migration.EN 10088, Stainless steels.EN ISO 8442-2:1997, Materials and articles in contact with foodstuffs – Cutlery and table holloware – Part 2:Requirements for stainless steel and silver-plated cutlery (ISO 8442-2:1997).ISO 648, Laboratory glassware - One mark pipettes.ISO 4788, Laboratory glassware - Graduated measuring cylinders.3 PrincipleThe overall migration from a sample of the plastics is determined as the loss in mass per unit of surface areaintended to come into contact with foodstuffs.The selection of the conditions of test will be determined by the conditions of use, see clauses 4, 5 and 6 of EN1186-1:2002.Test specimens of known mass are immersed in olive oil for the exposure time, at temperatures above 20 °C andbelow 100 °C, then taken from the olive oil, blotted to remove oil adhering to the surface, and reweighed.The specimens will usually retain absorbed olive oil that is extracted and determined quantitatively by means of gaschromatography after conversion to methyl esters.
Methylation is carried out by reacting a borontrifluoride/methanol complex with fatty acids formed by hydrolysing the oil with potassium hydroxide.
An internalstandard, triheptadecanoin, is added prior to the extraction of the absorbed olive oil from the test specimens.
ThisSIST EN 1186-2:2002
If an unacceptable amount of interference is present thensuitability of one of the 'other fatty food simulants' should be examined, see annex A and 9.3 and 9.5 of EN 1186-1:2002.
If aninterference is present which would interfere with the triheptadecanoin internal standard an alternative internal standard shouldbe used, see annex A and 9.3 of EN 1186-1:2002.4 ReagentsNOTEAll reagents should be of recognized analytical quality, unless otherwise specified.4.1Olive oil, reference simulant D, as specified in 5.2 of EN 1186-1:2002.4.2Extraction solvent (see 10.1 of EN 1186-1:2002).4.2.1For non-polar plastics, such as polyethylene and polypropylene:- Pentane 98 % boiling point 36 °C.For polar plastics, such as polyamide and polyacetal:- 95/5 by volume azeotropic mixture of pentane 98 % and ethanol 99 %.NOTE 1Pentane is a very volatile and highly flammable solvent.
Care should therefore be taken when handling this solventto prevent contact with sources of ignition.
Ethanol is also a flammable solvent.
It is not recommended that extractions witheither pentane or the pentane/ethanol mixture be left unattended, particularly overnight.NOTE 2Due to the low boiling points of these solvents, cooled condenser water can be required to prevent undue loss of thesolvent from the condenser.4.2.2Other suitable solvent.NOTE 1In previous methods for determining overall migration into olive oil the extraction solvent used has been 1,1,2-trichloro-trifluoroethane.
For environmental reasons the use of this solvent should be avoided where possible, see 9.1 of EN1186-1.
Experience has shown that this solvent, although effective for most plastics requires longer periods of extraction.NOTE 2Some solvents can contain non-volatile substances which, after hydrolysis and methylation processes, produce gaschromatography peaks with retention times similar to the retention times of olive oil methyl esters and methyl heptadecanoatefrom the internal standard.
Solvents found to contain such substances should be redistilled before use.SIST EN 1186-2:2002

Before initial use thoroughly clean the steel supports. The use of a degreasing solvent andthen dilute nitric acid has been found to be suitable.NOTEThe method has been written for the supports shown in Figure C.1 of EN 1186-1:2002 which have been found to besuitable for holding thin film and sheet test pieces.
However other supports can be used providing they are capable of holdingand keeping the test pieces apart and at the same time ensuring complete contact with the simulant.
For rigid samples,supports with a single cross arm can be used.5.9Gauze, pieces of fine stainless steel gauze, with a mesh size of 1 mm have been found to be suitable,approximately 25 mm ´ 100 mm for insertion between the test pieces on the supports.
Before initial use thoroughlyclean the gauze, first with a degreasing solvent and then with dilute nitric acid.
1) The source of this is the Chemical Abstracts published by the American Chemical Society.SIST EN 1186-2:2002

Therefore thecontainers should be placed in a thermostatically controlled room or oven, at a temperature of approximately 20 °C, the settemperature should not vary by more than ± 1 °C.5.11 Glass tubes, ground neck and stoppers, for retaining the olive oil and test specimens. Tubes with an internaldiameter of approximately 35 mm and length in the range of 100 mm to 200 mm, excluding the ground neck, see7.2 of EN 1186-1:2002, have been found to be satisfactory.5.12Thermostatically controlled oven or incubator capable of maintaining the set temperature, within thetolerances specified in Table B.2 of EN 1186-1:2002.5.13Filter paper, lint-free.5.14Anti-bumping beads.5.15Soxhlet type extractors, capable of holding test specimens on the supports, with 250 ml or 500 ml roundbottom flasks to fit.NOTEAlternative extractors capable of satisfactorily extracting absorbed olive oil from the test specimens can be used.5.16Water bath, capable of holding the flasks of soxhlet type extractors (5.15)5.17Rotary evaporator or distillation apparatus, for evaporation and collection of the extraction solvent.NOTEArtificially cooled water can be necessary for efficient condensation of a low boiling point solvent.5.18Steam bath or water bath.5.19Flasks, 50 ml, long neck with condensers to fit, for methyl ester preparations.5.20Measuring cylinders, complying with the minimum requirements of ISO 4788, 500 ml, 250 ml, 100 ml, 25 ml,and 10 ml.
A 10 ml graduated syringe may be used in place of the 10 ml measuring cylinder.5.21Pipettes, complying with the minimum requirements of ISO 648, 5 ml and 10 ml.5.22Glass beads, 2 mm to 3 mm in diameter or glass rods, 2 mm to 3 mm in diameter and approximately100 mm long (see 7.2 of EN 1186-1:2002).5.23Gas chromatograph, with flame ionisation detector equipped with an appropriate column.
When using apolar column, the major peaks of olive oil, such as C16:0, methyl hexadecanoate (methyl palmitate), C16:1, methyl9-hexadecenoate (methyl palmitoate), C18:0, methyl octadecanoate (methyl stearate), C18:1, methyl 9-octadecenoate (methyl oleate), C18:2, methyl 9,12-octadecadienoate (methyl linoleate) and the internal standardC17:0, methyl heptadecanoate (methyl margarate) shall demonstrate baseline separation. Optionally, a non-polarcolumn can be used which shall give baseline separation of the methyl esters with 16 and 18 carbon numbers andthe internal standard with 17 carbon number.NOTEThe following columns have been found to be suitable:- Column 1, polar column, WCOT fused silica column, length 50 m, internal diameter 0,25 mm, coated with a 0,21micrometre film of cyanopropyl silicone;- Column 2, non polar column, BP1, length 25 m, internal diameter 0,32 mm, with a 1 micron film thickness;- Column 3, polar column, stainless steel column 2 mm to 3 mm internal diameter and 2 m to 3 m length with a packing of10 % to 20 % by mass of polyestersuccinate on a stationary phase of diatomaceous earth 80 mesh to 100 mesh.SIST EN 1186-2:2002

The vacuumoven or vacuum desiccator shall be equipped with or connected to a vacuum pump capable of achieving a vacuumof 1,3 kPa or less.
The vacuum pump shall be provided with a time controller to switch on the vacuum pump everyhour for 15 min.NOTEIf a vacuum oven is not available, a vacuum desiccator placed in an oven at 60 °C can be used.5.26Desiccator containing self indicating silica gel or anhydrous calcium chloride.5.27Balance, capable of determining a change of mass of 10 mg.5.28Disposable plastic syringes with luer fitting. 1 ml or 10 ml size.5.29Wide gauge luer needles (80 mm ´ 1.2 mm).5.30Karl Fischer apparatus, either an automated volumetric titrator, or an automated coulometric titrator.
TheKarl Fischer titrator shall be capable of measuring the water content of the simulant with a precision
(standarddeviation) of 10 mg/kg or less (equivalent to 1 mg/dm2 plastic).
An automated volumetric or coulometric instrumentshall be used.
Manual titration procedures do not give the required accuracy or precision.6 Preparation of test specimens6.1 GeneralIt is essential that test specimens are clean and free from surface contamination (many plastics can readily attractdust due to static charges). Before preparing test specimens, remove any surface contamination from the sampleby gently wiping it with a lint-free cloth, or by brushing with a soft brush. Under no circumstances wash the samplewith water or solvent. If it is specified in the instructions for use of the article that it should be washed or cleanedbefore use see 8.1 of EN 1186-1:2002.
Minimise handling of the samples and, where necessary, wear cottongloves.To ensure that test pieces are well separated and that their surfaces are freely exposed to olive oil during theperiod of the test, for thin films insert a piece of fine stainless steel gauze (5.9) between the test pieces or for thicksamples not placed on the supports, insert glass rods between the test pieces after immersion in the olive oil.Where specimen supports are used, label the supports with a tag bearing the test specimen identification.When preparing test specimens measure the surface area according to 8.3 of EN 1186-1:2002.6.2 Number of test specimensSeven test specimens are required for samples, in the form of thin films, sheets, cut sections from containers orsimilar articles.
Nine test specimens, similar dimensionally one to another, are required for samples of articles ofirregular shape.These test specimens are utilized as follows:a) four test specimens for the migration test;b) two test specimens to check for possible loss of volatiles;c) one test specimen to determine the suitability of olive oil as the fatty food simulant and triheptadecanoinas the internal standard (see annex A);SIST EN 1186-2:2002

If the vacuum drying procedure in annex C is used these test specimens are notrequired as during the vacuum drying any volatiles will have been removed from the test specimens.If previous testing has established that interference in the gas chromatography procedure is unlikely and annex A isomitted, one fewer test specimen will be required.A minimum of three valid test results is required to calculate the mean.
Testing in triplicate is allowed but in thiscase if one test result is invalid repeat the entire procedure.6.3 Films and sheetsLay the sample on the cutting slab (5.1) and cut test specimens of 1 dm², see 8.3 of EN 1186-1:2002, using the100 mm ´ 100 mm template (5.4).
Check, using the rule (5.6), that the dimensions of the test specimen are withinthe specified deviation (± 1 mm).Cut each test specimen into four test pieces 25 mm ´ 100 mm using the rule (5.5).
Assemble one test specimenonto the support by piercing suitable holes in the test pieces and placing two test pieces on each side of the crossarms of the support. Repeat this procedure for all remaining test specimens.6.4 Containers and other articlesCut sections from the walls of the container or article to give test specimens each of area approximately 1 dm².
Forarticles with individual areas less than 1 dm², use a number of articles to provide each test specimen.Measure the dimensions of each test specimen to the nearest 1 mm, using the rule, see 8.3 of EN 1186-1:2002.Calculate the area of each test specimen to the nearest 0,01 dm² and record. If necessary, cut each test specimeninto smaller pieces to enable them to fit into the glass tubes (5.11). The test specimens or pieces are placed on thespecimen supports if these are appropriate or, if the test specimens or pieces are sufficiently rigid, they can betested unsupported.NOTECutting the test specimens into smaller pieces will increase the area of cut edges, so that the area of cut edgesexceeds 10 % of the of the test specimen area.
In this case see 8.3 of EN 1186-1:2002.6.5 Articles of irregular shapeSelect representative portions of the article, or multiples of the article for small articles, to give nine dimensionallysimilar test specimens each with a known total surface area of at least 1 dm².
Measure only the surface areaintended to come into contact with foodstuffs of two of these test specimens to the nearest 0,05 dm² using theSchlegel Method, as described in EN ISO 8442-2:1997, annex B, or any other suitable method.
Record the surfacearea of each test specimen.7 Procedure7.1 GeneralDetermine the applicability of the method by carrying out the procedure described in annex A.
If prior tests haveestablished that the method is applicable then annex A may be omitted.Before weighing, discharge any build up of static electricity with an antistatic gun or other suitable means.SIST EN 1186-2:2002

If prior tests have established that sample conditioning is not required then annex B andannex C
may be omitted.
If prior tests have established that the procedure described in annex D is applicable tothe sample, then annex B or annex C may be omitted.7.2.2If the tests described in annex B or annex C show that conditioning is not necessary, determine and recordthe mass of each test specimen.7.2.3If the tests described in annex B or annex C show that conditioning is necessary, follow the directions inthe relevant annex to determine the initial mass of the sample.7.2.4If the tests described in annex B show that conditioning is necessary, but constant mass cannot beachieved within five days then carry out the conditioning procedure described in C.3.1 or annex D.NOTE 1Long conditioning periods are not satisfactory due to oxidation of the olive oil which can occur upon prolongedconditioning.NOTE 2The conditioning procedures described in annex C and annex D may be used if it has been established that theseprocedures are more suited to the
polymer type under test.7.3 Exposure to food simulantTake six of the glass tubes (5.11), mark them for identification purposes.
Measure 100 ml ± 5 ml of olive oil (4.1)into each tube by measuring cylinder and stopper the tube.NOTE 1If the procedure described in annex D is used, it can be necessary to dry all of the olive oil used for the migrationtest, see D.3.2.Alternatively mark the tubes for a volume of 100 ml and fill with olive oil to the mark.
Place into one of the tubes athermometer or thermocouple and stopper the tubes.
Two extra tubes with a minimum of 50 ml of olive oil arerequired as blank simulant, if the procedure in annex D is used.
Place the six or eight tubes, and two empty tubes,in the thermostatically controlled oven or incubator (5.12) set at the test temperature.
Leave until the olive oil hasattained the test temperature, using the thermometer or thermocouple to monitor the temperature.
Take all tubesfrom the oven and place into four of the tubes containing olive oil, weighed test specimens prepared as in clause 6and conditioned if necessary.
Stopper the tubes.
Ensure that the test specimens are totally immersed in olive oil; ifthey are not, then add either glass beads or glass rods (5.22) to raise the level of the olive oil until total immersionis achieved.NOTE 2The olive oil in the fifth tube is used as a reference standard in constructing the calibration graph (see 7.6.2.2) and ifthe procedure in annex D is used, as the third blank sample for Karl Fischer titrations.
The olive oil in the sixth tube is used tocheck the temperature of the oil.
If glass beads or glass rods have been used to raise the level of the olive oil to achieve totalimmersion, then similar glass beads or glass rods should be added to the sixth tube.Place the remaining two test specimens into the empty tubes and stopper.NOTE 3These two test specimens are used to check whether the sample losses mass from the evaporation of volatiles,such as water, solvents and oligomers, during the test period.
If the vacuum drying procedure in annex C is applicable thesetest specimens are not required as during the vacuum drying volatiles will have been removed from the test specimens.Replace all eight or ten tubes in the thermostatically controlled oven or incubator set at the test temperature.
Thispart of the operation should be carried out in the minimum time possible to prevent undue heat loss.
Observe thetemperature of the thermostatically controlled oven or incubator or the olive oil
(see NOTE 5) in the sixth tube andleave the tubes for the selected test period, taking into account the tolerances specified in Table B.1 of EN 1186-1:2002, after the olive oil in the sixth tube has reached a temperature within the tolerance specified in Table B.2 ofEN 1186-1:2002.NOTE 4Annex B of EN 1186-1:2002 includes tolerances on a wide range of contact times and contact temperatures.
All ofthese contact times and contact temperatures are not necessarily relevant to this Part of the standard.SIST EN 1186-2:2002

For thosespecimens that have been in olive oil, allow the oil to drain.
Remove any adhering olive oil by gently pressingbetween filter papers (5.13).
Repeat the pressing procedure until the filter paper shows no spots of olive oil.
Fortest specimens on supports, remove the individual test pieces from the supports to carry out this operation.
Cleanthe supports of oil by washing with the extraction solvent and replace the test pieces on them.NOTE 6If the procedure in annex D is followed, the tubes containing the oil should be retained. The tubes should be cappedto prevent further change in the moisture content of the oil and the Karl Fischer determination should be carried out as soon aspossible.7.4 Final weighing of test specimens7.4.1For those specimens which did not require conditioning to obtain their initial masses (see 7.2.2), weigh allsix test specimens i.e. the four that have been in olive oil and the two that were in the empty tubes and record themass of each test specimen.7.4.2If conditioning of the test specimens was carried out using the procedure in annex B (see 7.2.3) thenrepeat the procedure.7.4.3If conditioning was carried out before the initial weighing using the procedure described in annex C (see7.2.4) then carry out the procedure described in C.4.7.4.4If it was decided that the procedure described in annex D (see 7.2.4) was applicable to the test sample,then carry out that procedure.7.4.5If the final mass of each of the test specimens which have been in empty tubes is less than their initialmass by more than 2,0 mg, then volatile substances have been lost and adjustment may be made, see 9.5 of EN1186-1:2002, to the final mass for each test specimen such that the values obtained are a measure of the migrationof non-volatile substances only.7.5Extraction of absorbed olive oilTake four flasks, 250 ml or 500 ml as appropriate to the size of the soxhlet type extractor (5.15) to be used for theextraction, and place in each flask 10,0 ml of the internal standard cyclohexane solution of triheptadecanoin (4.3),using a pipette (5.21), or an alternative higher quantity if more than 100 mg of olive oil is present.NOTE 1If the test specimens have retained more than 100 mg of olive oil, 10,0 ml of the internal standard solution will beinsufficient for optimum precision in the gas chromatography determination after extraction.
Before commencing the operationsin this clause an estimation of the quantity of olive oil retained in the test specimens should be obtained by comparing the finalmasses of the test specimens with their initial masses.
If considered necessary the quantity of internal standard solution can beincreased from 10 ml although it is essential that the same quantity is used for each test specimen, and that this quantity is alsoused with the olive oil standards for the calibration graph (see 7.6.2.2).
As a guide, approximately 0,5 mg of the internalstandard is required for every mg of extracted olive oil.Add sufficient extraction solvent (4.2) to allow cycling of the soxhlet type extractor (approximately 200 ml or 400 ml,according to the size of the flask) with anti-bumping beads (5.14) to control boiling.Place the four test specimens that have been in contact with olive oil into four soxhlet type extractors.
Couple eachsoxhlet to a flask containing the internal standard prepared as above.
Using either a water bath or steam bath(5.16), extract for a period of 7 10+ h, with a minimum of six cycles per hour, ensuring that the test pieces are totallysubmerged in the solvent during each soxhlet cycle, and that they remain separated from each other.Drain all of the solvent from the soxhlet type extractors, remove the flasks from the soxhlet type extractors andevaporate the solvent to approximately 10 ml using a rotary evaporator, or simple distillation apparatus (5.17).Transfer the solutions containing the extracted olive oil and internal standard to separate 50 ml flasks (5.19), andwash each flask with three portions of 5 ml of solvent.
Add the three washings to the respective individual 50 mlflasks.
Evaporate to dryness using a rotary evaporator or a water bath (5.17 or 5.18).SIST EN 1186-2:2002

In these cases extraction of the olive oilis incomplete and a second extraction with a more polar solvent is required, see also 9.2 of EN 1186-1:2002.Repeat the extraction of the test specimens for an additional 710+ h, with diethyl ether (4.8), adding a furtherquantity of the internal standard solution.NOTE 4The same quantity of internal standard solution is used as for the first 7 h extraction.
This quantity may not be theoptimum if the quantity of olive oil in the first 7 h extraction is high. Good precision is not required for the second 7 hdeterminations since they are intended primarily as a check on the efficiency of the first 7 h extraction and using the samequantity of internal standard enables one calibration graph to be used.If previous testing has established that all of the olive oil will be extracted from the test specimens during the first 7h extraction then the second 7 h extraction may be omitted.Isolate the residues in 50 ml flasks, using the procedure described above.Determine the extracted olive oil in both the first 7 h and the second 7 h extraction by the procedure described in7.6, but retain the test specimens in the soxhlet type extractors until the extracted olive oil has been determined forthe second extraction.7.6.1Preparation of fatty acid methyl estersAdd 10 ml ± 0,2 ml of n-heptane to each of the 50 ml flasks containing the first 7 h extraction residue, by measuringcylinder (5.20), ensuring that the residues of olive oil and plastics extractables dissolve or are well dispersed byshaking, warming or by ultrasonic treatment.NOTE 1Unless the residues in the flasks are dissolved or well dispersed in the n-heptane, quantitative hydrolysis ormethylation of the olive oil and of the internal standard might not be obtained under the conditions described particularly whenthese residues contain extractables from plastics in excess of 50 mg.
The internal standard might not react with the plasticsextractables to the same degree as does the olive oil and correct results for olive oil might not be obtained.Add by measuring cylinder or graduated syringe (5.20), 10 ml ± 0,2 ml of the potassium hydroxide solution (4.4)and a few anti-bumping beads (5.14).
Connect a condenser to the flask and boil the mixture under reflux for10 min ± 1,0 min.Add through the condenser by measuring cylinder, or graduated syringe (5.20), 5,0 ml ± 0,2 ml of the methanolsolution of boron trifluoride (4.5) and boil the mixture under reflux for 2 min ± 0,25 min.Cool to room temperature and add, by measuring cylinder (5.20), 15 ml to 20 ml of saturated sodium sulfatesolution (4.7.2) and shake well.
Then add further sodium sulfate solution until the liquid level reaches the neck ofthe flask.
Allow to stand until the phases have separated.NOTE 2The methyl esters for the subsequent gas chromatographic determination are in the upper, n-heptane, layer.Treat the residues from the second 7 h extraction as described above.If there will be a delay of more than 7 days in using a methyl ester solution for the gas chromatographicdeterminations, transfer the n-heptane layer to a small stoppered tube (5.24) containing solid anhydrous sodiumsulfate (4.7.1) and store in a refrigerator.SIST EN 1186-2:2002

5 °C to 190 °C and maintained at190 °C for 8 min.injector temperature220 °Cdetector temperature240 °CFor column 2 described in the note to 5.23 the following operating conditions have been found to besuitablecarrier gasheliumoven temperature250 °C isothermalinjector temperature320 °Cdetector temperature320 °CFor column 3 described in the note to 5.23 the following operating conditions have been found to besuitablecarrier gasnitrogen at 25 ml/minoven temperature185 °C to 195 °Cinjector temperature190 °C to 200 °Cdetector temperature190 °C to 200 °CUse an integrator to measure the area of each of the olive oil peaks and the internal standard.
Optionally a chartrecorder may be used to record the chromatogram and the height of the various peaks is measured.
In this caseonly the height of the internal standard and the major peak of olive oil (C18 or C18:1) shall be used forquantification of the amount of olive oil.NOTE 2The use of an integrator and measurement of the peak area is the preferred method.7.6.2.2Calibration graphWeigh a range of quantities of the blank reference olive oil which has been subjected to the same test conditionsas the test specimens into 50 ml flasks (5.19).
Weigh a range of olive oil quantities spanning the quantities of oliveoil in the first 7 h extractions, taking no fewer than four standards.Add 10,0 ml of the internal standard cyclohexane solution of triheptadecanoin (4.3) to each flask using a pipette(5.21), or the alternative quantity which has been added to the extraction flasks in 7.5.
Remove the cyclohexaneSIST EN 1186-2:2002

The following methods are acceptable:Method 1
Peak height methodMeasure the peak height of the internal standard peak and of the methyl oleate (C18:1 ) peak, when a polarcolumn has been employed.
In the case where a non-polar column has been used for the separation of the methylester, then measure the internal standard peak and the C18 peak of the olive oil.
Calculate the ratio of themeasured C18 peaks to the internal standard peak and plot the ratios versus the weighed quantities of olive oil.Method 2
Peak area methodMeasure the peak area of the internal standard peak and of each of the methyl esters originating from the olive oil.Add together the peak areas of the C16 and C18 peaks if a non-polar column was employed.
If a polar columnwas used, sum the areas of all the peaks (C16:0, C16:1, C18:0, C18:1 and C18:2) originating from the olive oil.Calculate the ratio of the combined areas of the measured peaks to the area of the internal standard peak and plotthe ratio versus the weighed quantities of olive oil.Method 3 Peak area method in the case of interference from the test sample.In the event that the analysis of a blank test sample, see annex A, has revealed an interference with one or more ofthe olive oil methyl esters, but not all
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