COMMISSION STAFF WORKING DOCUMENT Accompanying the document Proposal for a COUNCIL DECISION on the position to be taken on behalf of the European Union within the Council of Members of the International Olive Council (IOC), as regards one method of analysis and the IOC Trade Standard for olive oil and olive-pomace oil
Date: 2024-09-23
Institutions: Directorate-General for Agriculture and Rural Development, European Commission
Subjects: commodity agreement, international organisation, multilateral agreement, olive, olive oil, quality control of agricultural products

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  EN 

   EUROPEAN COMMISSION  Brussels, 23.9.2024  SWD(2024) 224 final  COMMISSION STAFF WORKING DOCUMENT   Accompanying the document Proposal for a COUNCIL DECISION on the position to be taken on behalf of the European Union within the Council of Members of the International Olive Council (IOC), as regards one method of analysis and the IOC Trade Standard for olive oil and olive-pomace oil {COM(2024) 416 final}  
 
 
DRAFT DECISION No DEC III.X/119-VI/2024 

CONCERNING THE METHOD FOR THE DETERMINATION OF  
THE CONTENT OF WAXES, FATTY ACID ETHYL ESTERS BY 
CAPILLARY GAS CHROMATOGRAPHY 

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DRAFT DECISION Nº DEC-III.X/119-VI/2024 

CONCERNING THE METHOD FOR THE DETERMINATION OF THE CONTENT OF WAXES, FATTY 
ACID ETHYL ESTERS BY CAPILLARY GAS CHROMATOGRAPHY 

THE COUNCIL OF MEMBERS OF THE INTERNATIONAL OLIVE COUNCIL, 

Having regard to the International Agreement on Olive Oil and Table Olives, 2015, in particular Article 1 
“Objectives of the Agreement” concerning standardisation and research, as regards achieving uniformity 
in  national  and  international  legislation,  and  the  harmonisation  of  physico-chemical  and  organoleptic 
analysis, to improve knowledge of the composition and quality characteristics of olive products, and in 
particular Chapter VI “Standardisation provisions”; 

Having  regard  to  the recommendation made  by  the  Chemistry  and  Standardisation Committee  at  its 
14th meeting during the 119th session of the Council of Members; 

Considering the ring tests carried out by the chemists on waxes and ethyl esters; 

Considering the unanimous position of the chemistry experts appointed by Members at their meeting 
on  3  and  4  April  2024,  and  the  recommendation  to  incorporate  the  modification  into  document 
COI/T.20/Doc. No 28/Rev. 4; 

DECIDES 

To adopt revision 4 of the document COI/T.20/Doc. No 28 on the method of analysis for the determination 
of the content of waxes and fatty acid ethyl esters by capillary gas chromatography. 

Both methods A and B can be used for official control. Method A (15 g of silica) is the reference method 
for counter-assessment. 

Document  COI/T.20/Doc.  No.  28/Rev.  4  June  2024  replaces  and  repeals  document  COI/T.20/Doc.  No 
28/Rev. 3 Corr 1 June 2024. 

Madrid (Spain), 25 June 2024 

Javad Mir Arabrazi 
Chair of the International Olive Council 

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INTERNATIONAL 

OLIVE 

COUNCIL 

COI/T.20/Doc. No 28/Rev. 3 4 
November 2023 June 2024 
page 1 

ENGLISH 
Original: ENGLISH 

Príncipe de Vergara, 154 – 28002 Madrid – Spain  Tel.: +34 915 903 638  Fax: +34 915 631 263 - E-mail: iooc@internationaloliveoil.org – http://www.internationaloliveoil.org/ 

METHOD OF ANALYSIS 

DETERMINATION OF THE CONTENT OF WAXES AND FATTY ACID ETHYL 
ESTERS BY CAPILLARY GAS CHROMATOGRAPHY 

METHOD A (15 g of silica) 

1. 

PURPOSE 

This method is for the determination of the content of waxes and fatty acid ethyl esters in olive oils. The 
individual waxes and alkyl esters are separated according to the number of carbon atoms. 

2. 

SCOPE 

The method is recommended as a tool for distinguishing between olive oil and olive-pomace oil, and as a 
quality parameter for extra virgin oils, as it facilitates the identification of false blends of extra virgin olive 
oils and low-quality oils and determines whether they are virgin, lampante or deodorized low quality oils 
that may have underwent a deodorization process. This document presents two methods that can be used 
both for official control. Method A (15 g of silica) is the reference method for counter-assessment. 

2. 3.  PRINCIPLE 

Addition of suitable internal standards to the oil and fractionation by chromatography on hydrated silica 
gel column. Recovery of the fraction eluted under the test conditions (with a lower polarity than that of the 
triacylglycerols)  and  direct  analysis  by  capillary  gas  chromatography.  Waxes  and  alkyl  esters  can  be 
analysed using either the reference method of part A or the alternative method of part B. Both methods 
share  the  same  principle,  but  the  simplified  method  uses  a  reduced  quantity  of  sample,  reagents,  and 
solvents. Method A is the reference method for counter assessment In case of disputes the results obtained 
using the reference method (part A) should prevail. 

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PART A – REFERENCE METHOD METHOD A – 15 GRAMS OF SILICA 

3. A4 -APPARATUS 

3.1. A4.1.  Test tube, 10 ml. 

3.2. A4.2.  Glass column for liquid chromatography, internal diameter 15 mm, length 40 cm, fitted with 

a suitable stopcock. 

3.3. A4.3.  Gas chromatograph suitable for use with a capillary column, equipped with a system for 

direct, on-column injection comprising: 

3.3.1. A4.3.1  Thermostat-controlled oven with temperature programming. 

3.3.2. A4.3.2  Cold injector for direct on-column injection 

3.3.3. A4.3.3  Flame ionisation detector and converter-amplifier. 

3.3.4. A4.3.4  Computerized  systems  recorder  Recorder-integrator  (Note  1)  for  use  with  the 

converter-amplifier (3.3.3), with a response time of not more than 1 s and a variable paper speed. 

3.3.5. A4.3.5  Capillary column, fused silica (for analysis of the waxes and methyl and ethyl 

esters), length 8-12 m, internal diameter 0.25-0.32 mm, internally coated with liquid phase (Note 2) 

to a uniform thickness of 0.10-0.25 m. 

3.4. A4.4.  Microsyringe, 10 l, with hardened needle, for direct on-column injection. 

3.5. A4.5.  Electric shaker. 

3.6. A4.6.  Rotary evaporator. 

3.7. A4.7.  Muffle oven. 

3.8. A4.8.  Analytical balance for weighing to an accuracy of ± 0.1 mg. 

3.9. A4.9.  Usual laboratory glassware. 

Note 1  Computerised systems may also be used where the gas chromatography data are entered through a PC. 
also to use with the converter-amplifier (A4.3.3) a recorder with a response time of not more than 1 s and 
a variable paper speed can be used. 
Suitable commercial liquid phases are available for this purpose such as SE52, SE54 (methyl silicon with 5% phenyl), 
etc. or other phase with similar or lower polarity. 

Note 2 

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4. A5 -REAGENTS 

4.1 A5.1. 

Silica gel, 60-200 m mesh. Place the silica gel in the muffle oven at 500 °C for at least 4 

hours. Allow to cool and then add 2% water in relation to the quantity of silica gel used. Shake well to 

homogenise slurry and keep in the desiccator for at least 12 hours prior to use. If the silica gel is ultra-pure 

grade, the muffle oven treatment will may not be necessary. In any case each batch of silica must be checked 

for suitability. 

4.2 A5.2. 

n-Hexane, chromatography (or residue) grade – the purity must be checked as follows: 100 

ml of n-hexane  are  evaporated to  dryness,  residue is  re-dissolved in  100  µl  n-heptane,  and analysed as 

solvent blank applying the same gas chromatographic conditions. There must be no peak in the elution 

alkyl esters area. (Hexane can be replaced by Isooctane) Hexane – Chromosolv Pestanal is available from 

Honeywell-Riedel-de Haen (code 34484). 

This reference is an example of suitable products, which are available commercially. This information is 

given  for  the  convenience  of  users  of  this  Standard  and  does  not  constitute  an  endorsement  of  these 

products. 

WARNING – Fumes may ignite. Keep away from sources of heat, sparks or naked flames. Make sure the 

bottles are always properly closed. Ensure proper ventilation during usage. Avoid build-up of fumes and 

remove any possible fire risk, such as heaters or electric apparatus not manufactured from non-inflammable 

material. Harmful if inhaled because it may cause nerve cell damage. Avoid breathing in the fumes. Use a 

suitable respiratory apparatus if necessary. Avoid contact with eyes and skin. 

4.3 A5.3. 

Ethyl ether, chromatography grade. 

WARNING – Highly inflammable and moderately toxic. Irritates the skin. Harmful if inhaled. May cause 

damage to eyes. Effects may be delayed. It can form explosive peroxides. Fumes may ignite. Keep away 

from sources of heat, sparks or naked flames. Make sure the bottles are always properly closed. Ensure 

proper ventilation during usage. Avoid build-up of fumes and remove any possible fire risk, such as heaters 

or electric apparatus not manufactured from non-inflammable material. Do not evaporate to dryness or near 

dryness. The addition of water or an appropriate reducing agent can reduce peroxide formation. Do not 

drink. Avoid breathing in the fumes. Avoid prolonged or repeated contact with skin. 

4.4 A5.4. 

n-hHeptane, chromatography grade, or iso-octane isooctane 

WARNING – Inflammable. Harmful if inhaled. Keep away from sources of heat, sparks or naked flames. 

Make sure the bottles are always properly closed. Ensure proper ventilation during usage. Avoid breathing 

in the fumes. Avoid prolonged or repeated contact with skin. 

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4.5 A5.5. 

Standard  solution  of  lauryl  arachidate  (Note  3)  at  0.02%  (m/V)  in  heptane  (internal 

standard for waxes). 

4.6 A5.6. 

Standard  solution  of  methyl  heptadecanoate  at  0.005%  (m/V)  in  heptane  (internal 

standard for methyl and ethyl esters). 

4.7 A5.7. 

Sudan  1  (1-phenylazo-2-naphthol)  optional  (attention:  azo-compounds  have  mutagenic 

and carcinogenic properties) 

4.8 A5.8. 

Carrier gas: hydrogen or helium, pure, gas chromatography grade. 

WARNING 

Hydrogen. Highly inflammable, under pressure. Keep away from sources of heat, sparks, naked flames 

or electric apparatus not manufactured from non-inflammable material. Make sure the bottle valve is 

shut when not in use. Always use with a pressure reducer. Release the tension of the reducer spring 

before opening the bottle valve. Do not stand in front of the bottle outlet when opening the valve. Ensure 

proper ventilation during usage. Do not transfer hydrogen from one bottle to another. Do not mix gas 

in the bottle. Make sure the bottles cannot be knocked over. Keep them away from sunlight and sources 

of heat. Store in a corrosive-free environment. Do not use damaged or unlabelled bottles. 

WARNING 

Helium. Compressed gas at  high  pressure.  It  reduces the amount  of oxygen available for breathing. 

Keep the bottle shut. Ensure proper ventilation during usage. Do not enter storage areas unless they are 

properly ventilated. Always use with a pressure reducer. Release the tension of the reducer spring before 

opening the bottle valve. Do not transfer gas from one bottle to another. Make sure the bottles cannot 

be knocked over. Do not stand in front of the bottle outlet when opening the valve. Keep them away 

from  sunlight  and  sources  of  heat.  Store  in  a  corrosive-free  environment.  Do  not  use  damaged  or 

unlabelled bottles. Do not inhale. Use solely for technical purposes. 

4.9 A5.9. 

Auxiliary gases: 

-  Hydrogen, pure, gas chromatography grade.  

-  Air, pure, gas chromatography grade. 

Note 3  Palmityl palmitate, myristyl stearate or arachidyl laureate may also be used. 

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WARNING 

Air. Compressed gas at high pressure. Use with caution in the presence of combustible substances as the 

self-ignition temperature of most of the organic compounds in the air is  considerably lower under high 

pressure. Make sure the bottle valve is shut when not in use. Always use a pressure reducer. Release the 

tension of the reducer spring before opening the bottle valve. Do not stand in front of the bottle outlet when 

opening the valve. Do not transfer gas from one bottle to another. Do not mix gas in the bottle. Make sure 

the bottles cannot be knocked over. Keep them away from sunlight and sources of heat. Store in a corrosive-

free environment. Do not use damaged or unlabelled bottles. Air intended for technical purposes must not 

be used for inhaling or respiratory apparatus. 

5. A6.  PROCEDURE 

5.1. A6.1. 

Preparation of the chromatography chromatographic column 

Suspend 15 g of silica gel (A4.5.1) in n-hexane (A4.5.2) and introduce into the column (A3.4.2). Allow to 

settle  spontaneously.  Complete  settling  with  the  aid  of  an  electric  shaker  (A3.4.5)  to  make  the 

chromatographic bed more homogeneous. Percolate 20 ml of n-hexane to remove any impurities. Weigh 

exactly, about 500 mg of the sample into the 10-ml test tube (A3.4.1), using the analytical balance (A3.4.8), 

and add a suitable amount of internal standard (A4.5.5) depending on the assumed wax content, e.g. add 

0.10 mg of lauryl arachidate in the case of extra virgin olive oil, virgin olive oil, refined olive oil and olive 

oil, 0.25-0.50 mg in the case of olive-pomace oil and 0.05 mg of methyl heptadecanoate for extra virgin 

olive oils (A4.5.6). Transfer the prepared sample to the chromatography column with the aid of two 2-ml 

portions of n-hexane (A4.5.2). Allow the solvent to flow to 1 mm above the upper level of the absorbent. 

Percolate a 50 ml n-hexane/ethyl ether (99:1) to further remove hydrocarbons (alkanes and sterenes). Then 

continue the chromatographic elution of n-hexane/ethyl ether (99:1) (Note 4) and collect 150 ml at a flow 

of about 15 drops every 10 seconds. (This fraction contains ethyl esters and waxes) (Note 5). 

Note 4 The n-hexane/ethyl ether (99:1) mixture should be freshly prepared every day, n-hexane can be replaced with 
the same amount of iso octane isooctane 
Note 5. 100 l of Sudan I dye at 1% in the elution mixture can be added to the sample solution to check visually that 
the waxes are eluted properly. The retention time of the dye lies in between that of the waxes and triacylglycerols. 
Hence, when the dye reaches the bottom of the chromatography column, elution must be suspended because all the 
waxes have been eluted. Verify the correct elution by checking the presence on the chromatogram at the same time 
of squalene and epoxy squalene. 
The elution volumes indicated in the text are those which usually are employed. However, if needed, they can be 
modified to achieve a satisfactory separation of the fractions. For this purpose, 100 μl of Sudan I dye at 1% in the 
elution mixture can be added to the sample solution to check visually that the waxes are eluted properly. The retention 
volume of the dye lies in between that of the waxes and triacylglycerols. Hence, when the dye reaches the bottom of 
the chromatography column, elution must be suspended because all the waxes have been eluted. Verify the correct 
elution by checking the presence on the chromatogram at the same time of squalene and epoxy squalene. 

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Evaporate  the  resultant  fractions  in  a  rotary  evaporator  (A3.4.6)  until  the  solvent  is  almost  removed. 

Remove the last 2 ml under a weak current of nitrogen. Collect the fraction containing the ethyl esters and 

waxes diluted with 1-2 ml of n-heptane or iso-octane. 

5.2. A6.2. 

Gas chromatography analysis 

5.2.1. A6.2.1.  Preliminary procedure 

Fit the column to the gas chromatograph (3.3), connecting the inlet port to the on-column system and the 

outlet port to the detector. Check the gas chromatography apparatus (operation of gas loops, efficiency of 

detector and recorder system, etc.). 

If the column is being used for the first time, it is advisable to condition it. Run a light flow of gas through 

the column, then switch on the gas chromatography apparatus. Gradually heat to 350 °C (approximately 4 

hours). 

Maintain  this  temperature  for  at  least  2  hours,  then  regulate  the  apparatus  to  the  operating  conditions 

(regulate gas flow, light flame, connect to electronic recorder (3.A.4.3.4), regulate oven temperature for 

column, regulate detector, etc.). Record the signal at a sensitivity at least twice as high as required for the 

analysis. The base line should be linear, with no peaks of any kind, and must not have any drift. 

Negative  straight-line  drift  indicates  that  the  column  connections  are  not  correct  while  positive  drift 

indicates that the column has not been properly conditioned. 

5.2.2. A6.2.2.  Choice of operating conditions for waxes and ethyl esters (Note 6) 

The operating conditions are generally as follows: 

-  Column temperature: 

  20°C/min         5°C/min 

80°C at first (1’)        140°C         335°C (20’) for ethyl esters and waxes 

80°C at first (1’)           240°C            325°C (6’)            340ºC (10’) for waxes only 

  20°C/min                5°C/min                 20°C/min 

Note 6 . Due to the high final temperature, positive drift is allowed but may not exceed more than 10% of the full-

scale value. 

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-  Detector temperature: 350°C. 

-  Amount injected: 1 µl of n-heptane solution (1-2ml). 

-  Carrier gas: helium or hydrogen at the optimal linear speed for the gas chosen (see Annex A B). 

- 

Instrument sensitivity: suitable for fulfilling the above conditions (a detection limit of about 1 

mg/Kg for each identified peak recommended) suitable for the quantification of the analytes. 

These conditions may be modified to suit the characteristics of the column and the gas chromatograph in 

order to separate all the waxes and fatty acid ethyl esters and to obtain satisfactory peak separation (see 

Figures1-and 2) and a retention time of 18  3 minutes for the lauryl arachidate internal standard. The most 

representative peak of the waxes must be over 60% of the full-scale value in the case of refined olive oil, 

olive oil, olive pomace oils, while it should be between 60% and full scale in the case of extra virgin olive 

oil and virgin olive oil. The methyl heptadecanoate internal standard for the ethyl esters must fit the full-

scale value. 

5.3. A6.3. 

Performance of the analysis 

Take up 1-2 µl of the solution with the aid of the 10 µl micro-syringe, drawing back the plunger 

until the needle is empty. Introduce the needle into the injection system and inject quickly after 1–

2 s. After about 5 s, gently extract the needle. 

Perform the recording until the waxes (C40-C46) are completely eluted, depending on the fraction 

being analysed. 

The base line must always meet the required conditions. 

5.4. A6.4. 

Peak identification 

Identify the peaks from the retention times by comparing them with mixtures of waxes with known 

retention times, analysed under the same conditions. The alkyl esters are identified from mixtures 

of methyl and ethyl esters of the main fatty acids in olive oils (palmitic and oleic). 

Figure 1. (Annex A) Sshows a chromatogram of the FAEE and waxes in an extra virgin olive oil 

using the method A (15 g). 

Figure 2. (Annex A) Sshows a chromatogram of the FAEE and waxes in a lampante olive oil using 

the method A (15 g). 

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5.5. A6.5. 

Quantitative analysis of the waxes 

Determine the area of the peaks corresponding to the lauryl arachidate internal standard and the 

aliphatic esters from C42 to C46 in the case of extra virgin olive oil and virgin olive oil and form 

C40 to C46 in the case of other oils, with the aid of the integrator. 

Determine the content of each individual wax, in mg/kg of fat, as follows: 

𝑊𝑎𝑥𝑒𝑠, 𝑚𝑔/𝑘𝑔 =

𝐴𝑥 ∗ 𝑚𝑠 ∗ 1000
𝐴𝑠 ∗ 𝑚

where: 

Ax = 

area corresponding to the peak for the individual ester, in computer counts (peak 

no. 11-13, 14-15, 16-17-18 in fig. 2 sum of peaks: C42, C44, C46 for EVOO and 

VOO; sum of peaks: C40, C42, C44 and C46 for the other oil) 

As   = 

area  corresponding  to  the  peak  for  the  lauryl  arachidate  internal  standard,  in 

computer counts 

ms  =  mass of the lauryl arachidate internal standard added, in milligrams 

m   =  mass of the sample taken for determination, in grams 

5.6. A6.6.Quantitative analysis of the ethyl esters 

Using  the  integrator,  determine  the  areas  of  the  peaks  corresponding  to  the  methyl  heptadecanoate 

internal standard, the ethyl esters of the C16 and C18 fatty acids. 

Determine the content of ethyl ester, in mg/kg of fat, as follows: 

𝐸𝑠𝑡𝑒𝑟, 𝑚𝑔/𝑘𝑔 =

𝐴𝑥 ∗ 𝑚𝑠 ∗ 1000
𝐴𝑠 ∗ 𝑚

where: 

Ax = 

area corresponding to all the peak for the individual C16 and C18 ethyl ester, in 

computer counts (sum of EtC16, EtC18:2, EtC18:1, EtC18). 

As   = 

area corresponding to the peak for the methyl heptadecanoate internal standard, in 

computer counts 

ms  =  mass of the methyl heptadecanoate internal standard added, in milligrams; 

m   =  mass of the sample taken for determination, in grams. 

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6. A7. 

EXPRESSION OF RESULTS 

Report the sum of the contents of the different waxes from C40 to C46 in milligrams per kilograms of fat 

for  Ordinary  Olive  oil  (OOO),  Lampante  olive  oil  (LOO),  Refined  olive  oil  (ROO),  Olive  oil 

(ROO+VOOs),  Crude  olive  pomace  oil  (COPO),  Refined  olive  pomace  oil  and  Olive  pomace  oil 

(ROPO+VOOs). Report the sum of the contents of the different waxes from C42 to C46 (Note  7) in the 

case  of  extra  virgin  and  virgin  olive  oils  and  from  C40  to  C46  in  the  case  of  other  oils  (Note  10)  in 

milligrams per kilograms of fat (ppm) in milligrams per kilograms of fat for Extra virgin Olive oil (EVOO) 

and Virgin Olive oil (VOO). Report the sum of the content of C40 to C46 in the case of the other categories 

of olive or olive pomace oils. 

Report the sum of the contents of the ethyl esters from C16 to C18 and the total of the two. 

Results should be expressed to the nearest mg/kg. 

Note 7. The components for quantification refer to the peaks with even carbon numbers amongst the C40-
C46 esters, according to the specimen chromatogram of the waxes in olive oil provided in the attached 
figure.  Peaks  identification  of  the  individual  waxes,  especially  if  C46  peak  is  splitted,  can  be  made  by 
comparing the retention time of the wax fraction of an olive-pomace oil. 

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METHOD B (3 g GRAMS OF SILICA) 

1. PURPOSE 

This method is for the determination of the content of waxes and fatty acid ethyl esters in olive oils. The 

individual waxes and alkyl esters are separated according to the number of carbon atoms. The method is 

recommended as a tool for distinguishing between olive oil and olive-pomace oil and as a quality parameter 

for extra virgin oils, as it facilitates the identification of false blends of extra virgin olive oils and low-

quality oils and determines whether they are virgin, lampante or deodorized oils. 

2. PRINCIPLE 

Addition of suitable internal standards to the oil and fractionation by chromatography on a hydrated silica 

gel column. Recovery of the fraction eluted under the test conditions (with a lower polarity than that of the 

triacylglycerols) and direct analysis by capillary gas chromatography. 

3. B4.  APPARATUS 

3.1. B4.1. 

Test tube, 10 ml. 

3.2. B4.2. 

Glass column for liquid chromatography, internal diameter 10 mm, length 40 cm, fitted 

with a suitable stopcock. 

3.3. B4.3. 

Gas chromatograph suitable for use with a capillary column, equipped with a system for 

direct, on-column injection comprising: 

3.3.1. B4.3.1.  Thermostat-controlled oven with temperature programming. 

3.3.2. B4.3.2.  Cold injector for direct on-column injection 

3.3.3. B4.3.3.  Flame ionisation detector and converter-amplifier 

3.3.4. B4.3.4.  Recorder  Computerized  systems  recorder-integrator  (Note  1  8)  for  use  with  the 

converter-amplifier (3.3.3), with a response time of not more than 1 s and a variable paper speed. 

Note 1 8. Computerised systems may also be used where the gas chromatography data are entered 

through a PC. also to use with the converter-amplifier (A4.3.3) a recorder with a response time of 
not more than 1 s and a variable paper speed can be used. 

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3.3.5. B4.3.5.  Capillary  column,  fused  silica  (for  analysis  of  the  waxes  and  methyl  and  ethyl 

esters), length 8-12 m, internal diameter 0.25-0.32 mm, internally coated with liquid phase (Note 2 9) 

to a uniform thickness of 0.10-0.25 µm. 

3.4. B4.4.  Microsyringe, 10 µl, with hardened needle, for direct on-column injection. 

3.5. B4.5. 

Electric shaker 

3.6. B4.6. 

Rotary evaporator 

3.7. B4.7.  Muffle oven 

3.8. B4.8. 

Analytical balance for weighing to an accuracy of ± 0.1 mg. 

3.9. B4.9. 

Usual laboratory glassware 

4. B5.  REAGENTS 

4.1. B5.1. 

Silica gel, 60-200 µm mesh. Place the silica gel in the muffle oven at 500°C for at least 4 

hours. Allow to cool and then add 2% water in relation to the quantity of silica gel used. Shake well 

to homogenise slurry and keep in the desiccator for at least 12 hours prior to use. If the silica gel is 

ultra-pure grade, the muffle oven treatment will may not be necessary. In any case each batch of 

silica must be checked for suitability. 

4.2. B5.2. 

n-Hexane, chromatography (or residue) grade the purity must be checked as follows 200 

100 ml of n-hexane are evaporated to dryness, residue is redissolved re-dissolved in 100 µl n-heptane and 

analysed as solvent blank applying the same gas chromatographic conditions. There must be no peak in the 

elution alkyl esters area. (Hexane can be replaced by Isooctane) Hexane – Chromosolv Pestanal is available 

from Honeyell-Riedel-de Haen (code 34484). 

This reference is an example of suitable products which are available commercially. This information is 
given  for  the  convenience  of  users  of  this  Standard  and  does  not  constitute  an  endorsement  of  these 
products. 
WARNING – Fumes may ignite. Keep away from sources of heat, sparks or naked flames. Make sure the 

bottles are always properly closed. Ensure proper ventilation during usage. Avoid build-up of fumes and 

remove any possible fire risk, such as heaters or electric apparatus not manufactured from non-inflammable 

material. Harmful if inhaled because it may cause nerve cell damage. Avoid breathing in the fumes. Use a 

suitable respiratory apparatus if necessary. Avoid contact with eyes and skin. 

Note  2  9. Suitable commercial liquid  phases are available for  this purpose such as SE52, SE54 (methyl 
silicon with 5% phenyl), etc. or other phase with similar or lower polarity. 

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4.3. B5.3. 

Ethyl ether, chromatography grade. 

WARNING – Highly inflammable and moderately toxic. Irritates the skin. Harmful if inhaled. May cause 

damage to eyes. Effects may be delayed. It can form explosive peroxides. Fumes may ignite. Keep away 

from sources of heat, sparks or naked flames. Make sure the bottles are always properly closed. Ensure 

proper ventilation during usage. Avoid build-up of fumes and remove any possible fire risk, such as heaters 

or electric apparatus not manufactured from non-inflammable material. Do not evaporate to dryness or near 

dryness. The addition of water or an appropriate reducing agent can reduce peroxide formation. Do not 

drink. Avoid breathing in the fumes. Avoid prolonged or repeated contact with skin. 

4.4. B5.4. 

n-hHeptane, chromatography grade, or iso-octane isooctane. 

WARNING – Inflammable. Harmful if inhaled. Keep away from sources of heat, sparks or naked flames. 

Make sure the bottles are always properly closed. Ensure proper ventilation during usage. Avoid breathing 

in the fumes. Avoid prolonged or repeated contact with skin. 

4.5. B5.5. 

Standard solution of lauryl arachidate (Note3 10), at 0.01% 0.002% (m/V) in heptane 

(internal standard for waxes). 

4.6. B5.6. 

Solution  of  methyl  heptadecanoate,  at  0.002%  0.0005%  (m/V)  in  heptane  (internal 

standard for methyl and ethyl esters). 

4.7. B5.7. 

Sudan 1 (1-phenylazo-2-naphthol) optional (attention: azo-compounds have mutagenic 

and carcinogenic properties) 

4.8. B5.8. 

Carrier gas: hydrogen or helium, pure, gas chromatography grade. 

WARNING 

Hydrogen. Highly inflammable under pressure. Keep away from sources of heat, sparks, naked flames or 

electric  apparatus  not  manufactured from non-inflammable material.  Make sure  the bottle valve  is  shut 

when  not  in  use. Always  use  with  a  pressure  reducer.  Release  the  tension  of  the  reducer  spring  before 

opening the bottle valve. Do not stand in front of the bottle outlet when opening the valve. Ensure proper 

ventilation during usage. Do not transfer hydrogen from one bottle to another. Do not mix gas in the bottle. 

Make sure the bottles cannot be knocked over. Keep them away from sunlight and sources of heat. Store in 

a corrosive-free environment. Do not use damaged or unlabelled bottles. 

Note 3.10 Palmityl palmitate, myristyl stearate or arachidyl laureate may also be used.  

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WARNING  -  Helium.  Compressed  gas  at  high  pressure.  It  reduces  the  amount  of  oxygen  available  for 

breathing. Keep the bottle shut. Ensure proper ventilation during usage. Do not enter storage areas unless 

they are properly ventilated. Always use with a pressure reducer. Release the tension of the reducer spring 

before opening the bottle valve. Do not transfer gas from one bottle to another. Make sure the bottles cannot 

be knocked over. Do not stand in front of the bottle outlet when opening the valve. Keep them away from 

sunlight  and  sources  of heat.  Store  in  a  corrosive-free  environment.  Do  not  use  damaged  or  unlabelled 

bottles. Do not inhale. Use solely for technical purposes. 

4.9. B5.9. 

Auxiliary gases: 

-  Hydrogen, pure, gas chromatography grade.  

-  Air, pure, gas chromatography grade. 

WARNING 

Air. Compressed gas at high pressure. Use with caution in the presence of combustible substances as the 

self-ignition temperature of most of the organic compounds in  the air is considerably  lower under high 

pressure. Make sure the bottle valve is shut when not in use. Always use a pressure reducer. Release the 

tension of the reducer spring before opening the bottle valve. Do not stand in front of the bottle outlet when 

opening the valve. Do not transfer gas from one bottle to another. Do not mix gas in the bottle. Make sure 

the bottles cannot be knocked over. Keep them away from sunlight and sources of heat. Store in a corrosive-

free environment. Do not use damaged or unlabelled bottles. Air intended for technical purposes must not 

be used for inhaling or respiratory apparatus. 

5. B6.  PROCEDURE 

5.1. B6.1. 

Preparation of the chromatography chromatographic column 

Suspend 3 g of silica gel (4 B5.1) in n-hexane (4 B5.2) and introduce into the column (3 B4.2). Allow 

to  settle  spontaneously.  Complete  settling  with  the  aid  of  an  electric  shaker  (3  B4.5)  to  make  the 

chromatographic  bed  more  homogeneous.  Percolate  10  ml  of  n-hexane  to  remove  any  impurities. 

Weigh  exactly, about  100 mg  of the sample into the 10-ml test tube  (3  B4.1), using the analytical 

balance (3 B4.8), and add a suitable amount of internal standard (4 B5.5) depending on the assumed 

wax content, e.g., add 0.01 0.02 mg (1 ml) of lauryl arachidate in the case of extra virgin olive oil, 

virgin olive oil, refined olive oil and olive oil, 0.025 0.05-0.10 mg in the case of olive-pomace oil and 

0.002 0.005 mg (1 ml) of methyl heptadecanoate for extra virgin olive oil and olive oil (4 6). Transfer 

the prepared sample to the chromatography column with the aid of two 2-1-ml portions of n-hexane 

(4 B5.2). 

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Allow the solvent to flow to 1 mm above the upper level of the absorbent. Percolate a 12-15 ml n-Hexane(*) 

to  further  remove  hydrocarbons  (alkanes  and  sterenes).  Then  start  the  chromatographic  elution  of  n-

hexane/ethyl ether (99:1) (Note 4) and collect 40-45 ml at a flow of about 15 drops every 10 seconds. 

This fraction contains the ethyl esters and waxes (Note 5 8 and Note 9). 

(*) Not necessary for the determination of waxes only. 

Evaporate  the  resultant  fractions  in  a  rotary  evaporator  (3  B4.6)  until  the  solvent  is  almost  removed. 

Remove the last 2 ml under a weak current of nitrogen. Collect the fraction containing the methyl and ethyl 

esters diluted with 0.5-1 ml of n-heptane or iso-octane isooctane. 

5.2. B6.2. 

Gas chromatography analysis 

5.2.1. B6.2.1. 

Preliminary procedure 

Fit the column to the gas chromatograph (3 B4.3), connecting the inlet port to the on-column system and 

the outlet port to the detector. Check the gas chromatography apparatus (operation of gas loops, efficiency 

of detector and recorder system, etc.). 

If the column is being used for the first time, it is advisable to condition it. Run a light flow of gas through 

the column, then switch on the gas chromatography apparatus. Gradually heat to 350 °C (approximately 4 

hours). 

Maintain  this  temperature  for  at  least  2  hours,  then  regulate  the  apparatus  to  the  operating  conditions 

(regulate gas flow, light flame, connect to electronic recorder (3.3.4), regulate oven temperature for column, 

regulate  detector,  etc.).  Record  the  signal  at  a  sensitivity  at  least  twice  as  high  as  that  required  for  the 

analysis. The base line should be linear, with no peaks of any kind, and must not have any drift. Negative 

straight-line drift indicates that the column connections are not correct while positive drift indicates that 

the column has not been properly conditioned. 

Note4 11. The n-hexane/ethyl ether (99:1) mixture should be freshly prepared every day, n-hexane can be replaced 
with the same amount of iso-octane isooctane. 
Note5. 100 µl of Sudan I dye at 1% in the elution mixture can be added to the sample solution to check visually that 
the waxes are eluted properly. The retention time of the dye lies in between that of the waxes and triacylglycerols. 
Hence, when the dye reaches the bottom of the chromatography column, elution must be suspended because all the 
waxes have been eluted. 
Note12: The elution volumes indicated in the text are those which usually are employed. However, if needed, they can 
be modified to achieve a satisfactory separation of the fractions. For this purpose, 100 μl of Sudan I dye at 1% in 
the elution mixture can be added to the sample solution to check visually that the waxes are eluted properly. The 
retention volume of the dye lies in between that of the waxes and triacylglycerols. Hence, when the dye reaches the 
bottom of the chromatography column, elution must be suspended because all the waxes have been eluted. Verify the 
correct elution by checking the presence on the chromatogram at the same time of squalene and epoxy squalene. 

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5.2.2. B6.2.2.  Choice of operating conditions for waxes and ethyl esters (Note 6 13) 

The operating conditions are generally as follows: 

- 

Column temperature: 

20°C/min 

  5°C/min 

  80°C at first (1’)         140 °C          335°C (20’) for ethyl esters and waxes 

20°C/min 

    5°C/min 

  80°C at first (1’)              200°C          335°C (20’) for waxes only 

-  Detector temperature: 350°C. 

-  Amount injected: 1 µl of n-heptane solution (0.5-1ml). 

-  Carrier gas: helium or hydrogen at the optimal linear speed for the gas chosen (see Annex A B). 

- 

Instrument sensitivity: suitable for fulfilling the above conditions (a detection limit of about 

1 mg/Kg for each identified peak recommended) suitable for the quantification of the analytes. 

These conditions may be modified to suit the characteristics of the column and the gas chromatograph 

in order to separate all the waxes and fatty acid ethyl esters and to obtain satisfactory peak separation 

(see Figures1-2) and a retention time of 18  3 minutes for the lauryl arachidate internal standard. 

The most representative peak of the waxes must be over 60% of the full-scale value in the case of refined 

olive oil, olive oil, olive pomace oils, while it should be between 60% and full scale in the case of extra 

virgin olive oil and virgin olive oil. The methyl heptadecanoate internal standard for the ethyl esters must 

fit the full-scale value. 

-  Carrier gas: helium or hydrogen at the optimal linear speed for the gas chosen (see Annex A). 

- 

Instrument sensitivity: suitable for fulfilling the above conditions. 

Note 6.13 Due to the high final temperature, positive drift is allowed but may not exceed more than 10% of the full-
scale value. 

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5.3. B6.3. 

Performance of the analysis 

Take up 1-2 µl of the solution with the aid of the 10 µl micro-syringe, drawing back the plunger 

until the needle is empty. Introduce the needle into the injection system and inject quickly after 1–

2 s. After about 5 s, gently extract the needle 

Perform the recording until the waxes (C40-C46) are completely eluted, depending on the fraction 

being analysed. 

The base line must always meet the required conditions. 

5.4. B6.4. Peak identification 

Identify the peaks from the retention times by comparing them with mixtures of waxes with known 

retention times, analysed under the same conditions. The alkyl esters are identified from mixtures 

of methyl and ethyl esters of the chief main fatty acids in olive oils (palmitic and oleic).  

Annex A reports some examples of chromatograms of ethyl esters and waxes suitable to identify 

related peaks. 

Figure  3.  Shows  a  chromatogram  of  the  FAEE  and  waxes  in  an  extra  virgin  olive  oil  using  the 

method B (3 g). 

Figure 4. Shows the chromatograms of FAEE and waxes in a virgin olive oil using the method B (3 

g). 

5.5. B6.5. 

Quantitative analysis of the waxes 

Determine the area of the peaks corresponding to the lauryl arachidate internal standard and the 

aliphatic esters from C42 to C46 in the case of extra virgin olive oil and virgin olive oils and from 

C40 to C46 for in the case of other oils, with the aid of the integrator. 

Determine the content of each individual wax, in mg/kg of fat, as follows: 

𝑊𝑎𝑥𝑒𝑠, 𝑚𝑔/𝑘𝑔 =

𝐴𝑥 ∗ 𝑚𝑠 ∗ 1000
𝐴𝑠 ∗ 𝑚

where: 

Ax = 

area corresponding to the peak for the individual ester, in computer counts (peak n° 

11- 13, 14-15,16-17-18 in fig.2 sum of peaks: C42, C44, C46 for EVOO and VOO; 

sum of peaks: C40, C42, C44 and C46 for the other oil) 

As   = 

area  corresponding  to  the  peak  for  the  lauryl  arachidate  internal  standard,  in 

computer counts 

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ms =  mass of the lauryl arachidate internal standard added, in milligrams 

m =  mass of the sample taken for determination, in grams. 

5.6. B6.6. 

Quantitative analysis of the ethyl esters 

With the aid of Using the integrator, determine the areas of the peaks corresponding to the methyl 

heptadecanoate internal standard, the ethyl esters of the C16 and C18 fatty acids. 

Determine the content of ethyl ester, in mg/kg of fat, as follows: 

𝐸𝑠𝑡𝑒𝑟, 𝑚𝑔/𝑘𝑔 =

𝐴𝑥 ∗ 𝑚𝑠 ∗ 1000
𝐴𝑠 ∗ 𝑚

where: 

Ax = 

area corresponding to the peak for  all the individual C16 and C18 ethyl ester, in 

computer counts (sum of EtC16, EtC18:2, EtC18:1, EtC18). 

As   = 

area corresponding to the peak for the methyl heptadecanoate internal standard, in 

computer counts. 

ms  =  mass of the methyl heptadecanoate internal standard added, in milligrams. 

m   =  mass of the sample taken for determination, in grams. 

6. B7.  EXPRESSION OF RESULTS 

Report the sum of the contents of the different waxes from C42 to C46 in the case of extra virgin olive oil 

and  virgin  olive  oil  and from  C40  to  C46  in  the case  of  other  categories  of  olive  or  olive  pomace  oils 

(Note 7) in milligrams per kilograms of fat (Note 14). 

Report the sum of the contents of the ethyl esters from C16 to C18 and the total of the two. 

Results should be expressed to the nearest mg/kg. 

Note 7.14 The components for quantification refer to the peaks with even carbon numbers amongst the C40-C46 esters, 
according  to  the  specimen  chromatogram  of  the  waxes  in  olive  oil  provided  in  the  attached  figure.  Peaks 
identification of the individual waxes, especially if C46 peak is splitted, can be made by comparing the retention time 
of the wax fraction of an olive-pomace oil. 

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ANNEX A 

Examples of chromatograms: 

The following chromatograms are reported as an aid to identify peaks as well as to give information about 

the separation to be obtained. 

Figure 1: Chromatogram of FAEE and waxes of EVOO using method A (15 g). 

Figure 2: Chromatogram of FAEE and waxes of Lampante olive oil using method A (15 g). 

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Figure 3: Chromatogram of FAEE and waxes of EVOO using method B (3 g). 

Figure 4: Chromatogram of FAEE and waxes of VOO using method B (3 g). 

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ANNEX B 

Determination of linear gas speed 

Inject 1:3 μl of methane (or propane) into the gas chromatograph after adjusting it to the normal operating 

conditions. Measure the time the gas takes to run through the column from the moment it is injected until the 

peak emerges (tM). 

The linear speed in cm/s is given by L/tM where L is the length of the column, in cm, and tM is the time 

measured in s. 

ANNEX C 

Precision values of the ethyl esters and wax method  

Analysis of the collaborative test results  

The results of the collaborative test organised by the IOC Executive Secretariat were statistically processed 

according to the rules laid down in the international standards ISO 5725. 

Accuracy (trueness and precision) of measurement methods and results. 

The test was performed on five samples. 

M1: EVOO to determine FAEE and waxes. 

M2: EVOO to determine FAEE and waxes. 

M3: VOO to determine FAEE and waxes. 

M4: LOO to determine FAEE and waxes. 

M5: OPO to determine waxes. 

Outliers  were  examined  by  applying  Cochran’s  and  Grubbs’s  test  to  the  laboratory  results  for  each 

determination (replicates a and b). 

The precision values of the method are given in the table overleaf. 

The table lists: 

n 

number of participating laboratories 

outliers 

number of laboratories with outlying values 

mean   

mean of the accepted results 

r 

value  below  which  the  absolute  difference  between  two  single  independent  test  results 

obtained with  the same method on identical  test material  in  the same laboratory by the 

same operator using the same equipment within short intervals of time may be expected to 

lie with a probability of 95% 

Sr 

repeatability standard deviation. 

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RSDr (%) 

repeatability coefficient of variation (Sr x 100 / mean) 

R 

Sr 

value below which the absolute difference between two single test results obtained with 

the same method on identical test material in different laboratories with different operators 

using different equipment may be expected to lie with a probability of 95%. 

reproducibility standard deviation  

RSDr (%) 

reproducibility coefficient of variation (Sr x 100 / mean) 

Ethyl esters (mg/kg) – Method A 15 g of silica  

M1 

10.00 

16 

3 

0.574 

5.74 

1.61 

0.759 

7.59 

2.12 

M2 

8.21 

16 

2 

0.330 

4.02 

0.92 

0.915 

11.15 

2.56 

M3 

36.97 

16 

0 

1.316 

3.56 

3.68 

3.720 

10.06 

10.42 

M4 

50.83 

16 

0 

1.934 

3.80 

5.41 

6.736 

13.25 

18.86 

Waxes (mg/kg) Method A 15 g of silica 

M1 

93.00 

16 

4 

0.610 

0.66 

1.71 

8.053 

8.66 

22.55 

M2 

45.17 

16 

1 

1.406 

3.11 

3.94 

4.879 

10.80 

13.66 

M3 

38.54 

16 

2 

1.315 

3.41 

3.68 

5.590 

14.51 

15.65 

M4 

M5 

323.17 

2350.72 

16 

1 

4.346 

1.34 

12.17 

16 

0 

35.728 

1.52 

100.04 

23.649 

247.180 

7.32 

66.22 

10.52 

692.10 

Sample 

Mean 

n 

outliers 

Sr 

RSDr (%) 

r 

SR 

RSDR (%) 

R 

Sample 

Mean 

n 

outliers 

Sr 

RSDr (%) 

r 

SR 

RSDR (%) 

R 

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Ethyl esters (mg/kg) -Method B  (3 g of silica) 

Sample 

Mean 

n 

Outliers 

Sr 

RSDr (%) 

r 

SR 

RSDR (%) 

R 

M1 

10.00 

15 

4 

M2 

8.15 

15 

4 

M3 

39.22 

15 

0 

0.570 

0.225 

1.234 

5.90 

1.65 

1.681 

14.00 

3.92 

2.76 

0.63 

0.991 

12.16 

2.77 

3.15 

3.46 

4.450 

11.34 

12.46 

M4 

52.46 

15 

0 

1.903 

3.63 

5.33 

4.691 

8.94 

13.13 

For waxes determination, for both methods A and B: 

M1, M2 and M3 (sum of waxes from C42 to C46); M4 and M5 (sum of waxes from C40 to C46) 

Waxes (mg/kg) Method A 15 g of silica 

Sample 

Mean 

n 

outliers 

Sr 

RSDr (%) 

r 

SR 

RSDR (%) 

R 

M1* 

93.00 

16 

4 

0.610 

0.66 

1.71 

8.053 

8.66 

22.55 

M2* 

45.17 

16 

1 

1.406 

3.11 

3.94 

4.879 

10.80 

13.66 

M3* 

38.54 

16 

2 

1.315 

3.41 

3.68 

5.590 

14.51 

15.65 

M4** 

M5** 

323.17 

2350.72 

16 

1 

4.346 

1.34 

12.17 

16 

0 

35.728 

1.52 

100.04 

23.649 

247.180 

7.32 

66.22 

10.52 

692.10 

* Sum C42-C46; ** Sum C40-C46 

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Waxes (mg/kg) Method B (3 g of silica) 

Sample 

Mean 

n 

Outliers 

Sr 

RSDr (%) 

r 

SR 

RSDR (%) 

R 

M1* 

90.45 

15 

2 

M2* 

45.53 

15 

1 

M3* 

38.81 

15 

2 

1.770 

2.306 

1.745 

1.96 

4.96 

15.287 

16.90 

42.80 

5.06 

6.46 

7.066 

15.52 

19.79 

4.50 

4.89 

8.236 

21.22 

23.06 

M4** 

M5** 

325.47 

2354.21 

15 

0 

7.758 

2.38 

21.72 

15 

0 

50.548 

2.15 

141.54 

23.614 

213.990 

7.26 

66.12 

9.09 

599.17 

* Sum C42-C46; ** Sum C40-C46 

ANNEX D 

References 

ISO  5725-1:1994 Accuracy  (trueness  and  precision)  of  measurement  methods  and  results  – Part  1: 
General principles and definitions 

ISO 5725-2:1994 2019  Accuracy (trueness and precision) of measurement methods and results – 
Part  2:  Basic  method  for  the  determination  of  the  repeatability  and  reproducibility  of  a  standard 
measurement method 

ISO 5725-5:1998  Accuracy (trueness and precision) of measurement methods and results – Part 5: 
Alternative methods for the determination of the precision of a standard measurement method 

ISO 5725-6:1994  Accuracy (trueness and precision) of measurement methods and results – Part 6: 
Use in practice of accuracy values 

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DECISION Nº. DEC-III. X/119-VI/2024 

REVISING THE TRADING STANDARD FOR OLIVE OIL AND OLIVE-POMACE OIL 

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DRAFT DECISION Nº. DEC-III. X/119-VI/2024 

REVISING THE TRADING STANDARD FOR OLIVE OIL AND OLIVE-POMACE OIL 

THE COUNCIL OF MEMBERS OF THE INTERNATIONAL OLIVE COUNCIL, 

Having regard to the International Agreement on Olive Oil and Table Olives 2015, and in particular its 
article 1 ‘Objectives of the Agreement’ on standardisation and research, concerning the standardisation of 
national and international legislation relating to the physico-chemical and organoleptic characteristics of 
olive oils, olive-pomace oils and table olives in order to avoid any obstacle to trade, and its Chapter VI 
‘Provisions concerning standardisation’; 

Having regard to the recommendation made by the Committee on Chemistry and Standardisation at its 
14th meeting during the 119th session of the Council of Members; 

Considering the work carried out by the chemists on the application of methods and studies on olive oils 
with non-standard parameters; 

Considering the unanimous position of the expert chemists appointed by Members, formulated at the expert 
chemists' meeting of 3 and 4 April 2024, on: 

-  The inclusion of the revision 11 of the method for the organoleptic assessment of virgin olive 

oil (T.20/Doc. No. 15/Rev. 11). 

-  The inclusion of revision 3 Corr 1 of the method of analysis for the determination of the content 
of  waxes  and  fatty  acid  ethyl  esters  by  capillary  gas  chromatography  (COI/T.20/Doc.  No 
28/Rev. 3 Corr. 1). 

DECIDES 

1. 

2. 

3. 

To adopt the Trade Standard for Olive Oils and Olive Pomace Oils COI/T.15/NC No. 3/Rev. 20, 
attached to this decision, which replaces and repeals the Trade Standard for Olive Oils and Olive 
Pomace Oils COI/T.15/NC No. 3/Rev. 19 of November 2022; 
Members shall, subject to their respective legislation, take all relevant measures to implement the 
adopted Standard and shall communicate these measures to the Executive Secretariat once they 
are implemented; 
Non-Member States interested in international trade in olive oils and olive-pomace oils are invited 
to take the adopted Standard into consideration and to adapt their regulations to the provisions of 
this Standard. 

Madrid (Spain), 25 June 2024 

Javad Mir Arabrazi 
Chair of the International Olive Council 

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INTERNATIONAL 

OLIVE 

COUNCIL 

COI/T.15/NC No 3/Rev. 19 20 
June 2024 

ENGLISH 
Original: FRENCH 

TRADE STANDARD APPLYING TO OLIVE OILS 

AND OLIVE POMACE OILS 

1. 

SCOPE 

This standard applies to olive oils and olive pomace oils that are the object of international trade or of 
concessional or food aid transactions. 

2. 

DESIGNATIONS AND DEFINITIONS 

2.1.  Olive oils 

2.1.1.  Virgin olive oils are oils which are obtained from the fruit of the olive tree (Olea europaea L.) solely 
by mechanical or other physical means under conditions, particularly thermal conditions, that do not 
lead  to  alterations  in  the  oil,  and  which  have  not  undergone  any  treatment  other  than  washing, 
decantation, centrifugation and filtration. Virgin olive oils shall be classified and designated as follows: 

2.1.1.1. Virgin olive oils fit for consumption as they are: 

(i) Extra virgin olive oil: virgin olive oil which has a free acidity, expressed as oleic acid, of not more 
than 0.80 grams per 100 grams and the other physico–chemical and organoleptic characteristics of 
which correspond to those fixed for this category in this standard. 

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page 2 

(ii) Virgin olive oil: virgin olive oil which has a free acidity, expressed as oleic acid, of not more than 
2.0 grams per 100 grams and the other physico–chemical and organoleptic characteristics of which 
correspond to those fixed for this category in this standard. 

(iii) Ordinary virgin olive oil: virgin olive oil which has a free acidity, expressed as oleic acid, of not 
more than 3.3 grams per 100 grams and the other physico–chemical and organoleptic characteristics 
of which correspond to those fixed for this category in this standard.1/ 

2.1.1.2. Virgin olive oils that must undergo processing prior to consumption: 

Lampante virgin olive oil: virgin olive oil which has a free acidity expressed as oleic acid, of more than 
3.3  grams  per  100  grams  and/or  the  physico–chemical  and  organoleptic  characteristics  of  which 
correspond to those fixed for this category in this standard. It is intended for refining or for technical 
use. 

2.1.2.  Refined olive oil: olive oil obtained from virgin olive oils by refining methods which do not lead to 
alterations in the initial glyceridic structure. It has a free acidity, expressed as oleic acid, of not more 
than  0.30  grams  per  100  grams  and  its  other  physico–chemical  and  organoleptic  characteristics 
correspond to those fixed for this category in this standard.2/ 

2.1.3.  Olive oil composed of refined olive oil and virgin olive oils: oil consisting of a blend of refined olive 
oil and virgin olive oils fit for consumption as they are. It has a free acidity, expressed as oleic acid, of 
not more than 1.00 gram per 100 grams and its other physico–chemical and organoleptic characteristics 
correspond to those fixed for this category in this standard. 

2.2.  Olive  pomace  oil3/  is  the  oil  obtained  by  treating  olive  pomace  with  solvents  or  other  physical 
treatments, to the exclusion of oils obtained by re-esterification processes and of any mixture with oils 
of other kinds. It is marketed in accordance with the following designations and definitions: 

2.2.1.  Crude olive pomace oil: olive pomace oil, the physico–chemical and organoleptic characteristics of 
which correspond to those fixed for this category in this standard. It is intended for refining for use for 
human consumption, or it is intended for technical use. 

____________ 
1/  This product may only be sold direct to the consumer if permitted in the country of retail sale. If not 
permitted,  the  designation  of  this  product  shall  comply  with  the  legal  provisions  of  the  country 
concerned. 

2/  This product may only be sold direct to the consumer if permitted in the country of retail sale. 
3/  Olive pomace oil cannot be sold with the designation or definition “olive oil”. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 3 

2.2.2.  Refined olive pomace oil: oil obtained from crude olive pomace oil by refining methods which do not 
lead to alterations in the initial glyceridic structure. It has a free acidity, expressed as oleic acid, of not 
more than 0.30 grams per 100 grams and its other physico–chemical and organoleptic characteristics 
correspond to those fixed for this category in this standard.1/ 

2.2.3.  Olive pomace oil composed of refined olive pomace oil and virgin olive oils: oil consisting of a blend 
of refined olive pomace oil and virgin olive oils fit for consumption as they are. It has a free acidity of 
not more than 1.00 gram per 100 grams and its other physico–chemical and organoleptic characteristics 
correspond to those fixed for this category in this standard.2/ In no case shall this blend be called "olive 
oil". 

3. 

PURITY CRITERIA 

The identity characteristics comprising the purity criteria shall be applicable to olive oils and olive 
pomace oils. 

The limits established for each criterion include the precision values of the attendant recommended 
method. 

3.1. 

Fatty acid composition as determined by gas chromatography 

(% m/m methyl esters): 

-  Myristic acid 
-  Palmitic acid 
-  Palmitoleic acid 
-  Heptadecanoic acid 
-  Heptadecenoic acid 
-  Stearic acid 
-  Oleic acid 
-  Linoleic acid 
-  Linolenic acid 
-  Arachidic acid 
-  Gadoleic acid (eicosenoic) 
-  Behenic acid 
-  Lignoceric acid 

__________ 

< 0.03 
7.00 - 20.00 
0.30 - 3.50 
< 0.40 
< 0.60 
0.50 - 5.00 
55.00 - 85.00 
2.50 - 21.00 
< 1.003/ 
< 0.60 
< 0.50 
< 0.20* 
< 0.20 

1/  This product may only be sold direct to the consumer if permitted in the country of retail sale. 
2/  The country of retail sale may require a more specific designation. 
3/  When an edible virgin olive oil exhibits 1.00<linolenic acid%≤1.40, then this oil is authentic, provided 

that App. β-sito/Campe content ≥ 24 and all other purity criteria lie within the official limit 

*  Limit raised to < 0.30 for olive pomace oils. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
3.2. 

Trans fatty acid content (% trans fatty acids) 

COI/T.15/NC No 3/Rev. 19 20 
page 4 

C18:1  T 
% 

C18:2 T + 
C18:3 T % 

< 0.05 
< 0.10 
< 0.20 
< 0.20 
< 0.20 
< 0.40 
< 0.40 

< 0.05 
< 0.10 
< 0.30 
< 0.30 
< 0.10 
< 0.35 
≤ 0.35 

-  Edible virgin olive oils 
-  Lampante virgin olive oil 
-  Refined olive oil 
-  Olive oil (ROO+VOOs)1 
-  Crude olive pomace oil 
-  Refined olive pomace oil 
-  Olive pomace oil (ROPO+VOOs)2 

3.3. 

Sterol and triterpene dialcohol composition 

3.3.1.  Desmethylsterol composition (% total sterols) 

-  Cholesterol 
-  Brassicasterol 
-  Campesterol 
-  Stigmasterol 
-  Delta 7 stigmastenol 
-  Apparent beta sitosterol: 

beta-sitosterol + delta-5-avenasterol 
+ delta-5-23-stigmastadienol +clerosterol 
+ sitostanol + delta 5-24-stigmastadienol 

< 0.5 
< 0.1* 
< 4.0** 
< campesterol in edible oils 
< 0.5*** 
> 93.0 

__________ 
1 Blend of refined olive oil and virgin olive oils 
2 Blend of refined olive pomace oil and virgin olive oils 

*  Limit raised to < 0.2 for olive pomace oils. 

**  An  extra  virgin  or  virgin  olive  oil  that  exhibits  4.0<  campesterol  %  ≤4.5  is  authentic  provided  that 
stigmasterol ≤1.4%, Δ7-stigmastenol ≤0.3% and all other parameters lie within the limits fixed in this 
standard. 

*** An olive oil or olive-pomace oil that exhibits 0.5< Δ7-stigmastenol % ≤0.8 is authentic provided that: 

a) app. β-sitosterol/campesterol≥28, ΔECN42 ≤│0.10│ (for extra virgin or virgin olive oil) 
b) app. β-sitosterol/campesterol≥28, ΔECN42 ≤│0.15│, stigmastadiene≤0.30 (for lampante virgin olive 

oil) 

c) app. β-sitosterol/campesterol≥28, ΔECN42≤│0.15│ (for refined olive oil or olive oil (ROO+VOOs)) 
d) stigmasterol ≤1.4%, ΔECN42 ≤│0.40│ (crude olive-pomace oil, refined olive-pomace oil or olive 

pomace oil (ROPO+VOOs)) 

In all the above cases, all other parameters lie within the limits fixed in this standard. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 5 

3.3.2.  Total sterol content 

(mg/kg) 

-  Virgin olive oils 
-  Refined olive oil 
-  Olive oil (ROO+VOOs) 
-  Crude olive pomace oil 
-  Refined olive pomace oil 
-  Olive pomace oil (ROPO+VOOs) 

3.3.3.  Erythrodiol and uvaol content 

(% total sterols) 

-  Edible virgin olive oils 
-  Lampante virgin olive oil 
-  Refined olive oil 
-  Olive oil (ROO+VOOs) 
-  Crude olive pomace oil 
-  Refined olive pomace oil 
-  Olive pomace oil (ROPO+VOOs) 

3.4.  Wax content 

(mg/kg) 

C42 + C44 + C46 (mg/kg) 
- 

Extra virgin olive oil and virgin olive oil 

C40 + C42 + C44 + C46 (mg/kg) 
-  Ordinary virgin olive oil 
Lampante virgin olive oil 
- 
- 
Refined olive oil 
-  Olive oil (ROO+VOOs) 
Crude olive pomace oil 
- 
- 
Refined olive pomace oil 
-  Olive pomace oil (ROPO+VOOs) 

__________ 

> 1000 

> 2500 
> 1800 
> 1600 

< 4.5 
< 4.51/ 
< 4.52/ 
< 4.5 
> 4.53/ 
> 4.5 
> 4.5 

< 150 

< 250 
< 3001/ 
< 350 
< 350 
> 3502/ 
> 350 
> 350 

1/  When the oil has a wax content between 300 mg/kg and 350 mg/kg it is considered a lampante virgin 
olive oil if the total aliphatic alcohol content is < 350 mg/kg or the erythrodiol + uvaol content is < 
3.5%. 

2/  When the oil has an erythrodiol + uvaol content of between 4.5 and 6 %, the erythrodiol content must 

be < 75 mg/kg. 

3/  When  the  oil  has  a  wax  content  between  300  mg/kg  and  350  mg/kg  it  is  considered  a  crude  olive 
pomace oil if the total aliphatic alcohol content is > 350 mg/kg and the erythrodiol + uvaol content is 
> 3.5%. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
3.5.  Maximum difference between the actual and theoretical ECN 42 triacylglycerol content (%) 

COI/T.15/NC No 3/Rev. 19 20 
page 6 

-  Edible virgin olive oils 
-  Lampante virgin olive oil 
-  Refined olive oil 
-  Olive oil (ROO+VOOs) 
-  Crude olive pomace oil 
-  Refined olive pomace oil 
-  Olive pomace oil (ROPO+VOOs) 

3.6. 

  Stigmastadiene content 

(mg/kg) 

-  Extra virgin olive oil and virgin olive oil 
-  Ordinary virgin olive oil 
-  Lampante virgin olive oil 

3.7.  Content of 2-glyceryl monopalmitate 

(%) 

≤ |0.20| 
≤ |0.30| 
≤ |0.30| 
≤ |0.30| 
≤ |0.60| 
≤ |0.50| 
≤ |0.50| 

< 0.05 
< 0.10 
< 0.50 

-  Edible virgin olive oils and olive oil (ROO+VOOs): 

C16:0 < 14.00%; 2P < 0.9% 
C16:0 > 14.00%, 2P < 1.0% 

-  Non-edible virgin olive oils and refined olive oils: 

C16:0 < 14.00%; 2P < 0.9% 
C16:0 > 14.00%, 2P < 1.1% 

-  Olive pomace oil (ROPO+VOOs) 
-  Crude and refined olive pomace oils  

< 1.2% 
< 1.4% 

3.8.  Unsaponifiable matter 

(g/kg) 

-  Olive oils 
-  Olive pomace oils 

< 15 
< 30 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
4.  QUALITY CRITERIA 

The limits established for each criterion and designation include the precision values of the attendant recommended method 

COI/T.15/NC No 3/Rev. 19 20 
page 7 

Extra 
virgin 
olive 
oil 

Virgin 
olive 
oil 

Ordinary 
virgin 
olive 
oil 

Lampante 
virgin 
olive 
oil* 

Refined 
olive 
oil 

Olive 
Oil 
(ROO+VO
Os) 

Crude 
olive 
pomace 
oil 

Refined 
olive  
pomace 
oil 

Olive 
pomace 
oil 
(ROPO+VO
Os) 

4.1 Organoleptic 
characteristics 

- odour and taste 

- median of defect 
- median of the fruity 
attribute 

- colour 

- aspect at 20oC for 
24 hours 

Me = 0.0 
Me > 0.0 

0.0 < Me < 3.5 
Me > 0.0 

3.5 <Me < 6.0** 

Me > 6.0 

acceptable 

good 

acceptable 

good 

Light yellow 

Light 
yellow to 
green 

Light yellow 
to brownish 
yellow 

Light yellow 
to green 

limpid 

limpid 

limpid 

limpid 

4.2. Free acidity 

< 0.80 

< 2.0 

< 3.3 

> 3.3 

< 0.30 

< 1.00 

no limit 

< 0.30 

< 1.00 

% m/m expressed in 
oleic acid 

4.3. Peroxide value 

< 20.0 

< 20.0 

< 20.0 

no limit 

< 5.0 

< 15.0 

no limit 

< 5.0 

< 15.0 

in milleq. peroxide 
oxygen per kg/oil 

* ..... It is not obligatory for the criteria in 4.1, 4.2 and 4.3 to be concurrent; one is sufficient. 
** ... Or when the median of the defect is less than or equal to 3.5 and the median of the fruity attribute is equal to 0.0. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 8 

4.  QUALITY CRITERIA (contd.) 

Extra 
virgin 
olive 
oil 

Virgin 
olive 
oil 

Ordinary 
virgin 
olive 
oil 

Lampante 
virgin 
olive 
oil 

Refined 
olive 
oil 

Olive 
Oil 
(ROO+VOOs) 

Crude 
olive 
pomace 
oil 

Refined 
olive 
pomace 
oil 

Olive 
pomace 
oil 
(ROPO+VOOs) 

4.4. Absorbency in ultra-violet 

- 270 nm (cyclohexane) / 
  268 nm (iso-octane) 
- K 
- 232 nm* 

< 0.22 

< 0.25 

< 0.01 
< 2.50** 

< 0.01 
< 2.60** 

< 0.30 

< 0.01 

< 1.25 

< 0.16 

< 1.15 

< 0.15 

< 2.00 

< 0.20 

< 1.70 

< 0.18 

4.5. Moisture and volatile matter 

< 0.2 

< 0.2 

< 0.2 

< 0.3 

< 0.1 

< 0.1 

< 1.5 

< 0.1 

< 0.1 

(% m/m) 

4.6. Insoluble impurities in light 

< 0.10 

< 0.10 

< 0.10 

< 0.20 

< 0.05 

< 0.05 

< 0.05 

< 0.05 

petroleum 
(% m/m) 

4.7. Flash point 

4.8. Trace metals 
(mg/kg) 
Iron 
Copper 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

< 3.0 
< 0.1 

> 120oC 

4.9. Fatty acid ethyl esters 

≤ 35 mg/kg 

(FAEEs) 

4.10. Phenols content 

See section 11.21 

* ..... This determination is solely for application by commercial partners on an optional basis. 
** ... Commercial partners in the country of retail sale may require compliance with these limits when the oil is made available to the end consumer. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 9 

5. 

FOOD ADDITIVES 

5.1. 

Virgin olive oils and crude olive pomace oil: 

none permitted 

5.2. 

Refined olive oil, olive oil (ROO+VOOs), refined olive pomace oil and olive pomace oil (ROPO+VOOs): 
alpha-tocopherol permitted to restore natural tocopherol lost in the refining process. 

Maximum level: According to the Good Manufacturing Practices (GMP) 

6. 

CONTAMINANTS 

6.1. 

Heavy metals 

The products covered by this standard shall comply with the maximum levels of the General Standard for 
Contaminants and Toxins in Food and Feed (CODEX STAN 193-1995). 

6.2. 

Pesticide residues 

The products covered by this standard shall comply with those maximum residue limits established by 
the Codex Alimentarius Commission for these commodities. 

6.3. 

Halogenated solvents 

-  Maximum content of each halogenated solvent 
-  Maximum content of the sum of all halogenated solvents 

0.1 mg/kg 
0.2 mg/kg 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 10 

7. 

HYGIENE 

7.1. 

7.2. 

It is recommended that the products intended for human consumption covered by the provisions of this 
standard be prepared and handled in accordance with the appropriate sections of the General Principles of 
Food Hygiene (CAC/RP 1-1969), and other relevant Codex texts such as Codes of Hygienic Practice and 
Codes of Practice. 

The products intended for human consumption should comply with any microbiological criteria established 
in  accordance  with  the  Principles  for  the  Establishment  and  Application  of  Microbiological  Criteria 
(CAC/GL 21-1997). 

8. 

PACKING 

Olive oils and olive pomace oils intended for international trade shall be packed in containers complying 
with  the  General  Principles  of  Food  Hygiene  recommended  by  the  Codex  Alimentarius  Commission 
(CAC/RCP 1 - 1969), and other relevant texts such as Codes of Hygienic Practice and Codes of Practice. 

The containers used may be: 

8.1. 

tanks, containers, vats, which permit the transportation in bulk of olive oils and olive pomace oils; 

8.2. 

8.3. 

metal drums, in good condition, hermetically-sealed, which should be internally covered with a suitable 
varnish; 

metal  tins  and  cans,  lithographed,  new,  hermetically-sealed,  which  should  be  internally  covered  with  a 
suitable varnish; 

8.4. 

demi-johns, glass bottles or bottles made of suitable macromolecular material. 

9. 

CONTAINER FILLING TOLERANCE 

The volume occupied by the contents shall under no circumstances be less than 90% of the capacity of the 
container, except in the case of tin containers with a capacity of, or less than, 1 L in which the volume 
occupied shall under no circumstances be less than 80% of the capacity of the container; this capacity is 
equal to the volume of distilled water at 20oC which the container can hold when full. 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 11 

10. 

LABELLING 

In addition to the appropriate sections of the Codex General Standard for the Labelling of Pre-packaged 
Foods  (CODEX  STAN  1-1985)  and  the  guidelines  applying  to  food  not  intended  for  direct  sale  to 
consumers, the specific provisions providing the following information shall be applied: 

10.1.  On containers intended for direct sale to consumers  

10.1.1.  Name of the product 

The labelling on each container shall indicate the specific designation of the product contained, complying 
in every way with the relevant provisions of this standard. 

10.1.1.1. Designations of olive oils: 

- Extra virgin olive oil 
- Virgin olive oil 
- Ordinary virgin olive oil1/ 
- Refined olive oil1/ 
- Olive oil (ROO+VOOs)2/ 

10.1.1.2. Designations of olive pomace oils: 
- Refined olive pomace oil1/ 
- Olive pomace oil (ROPO+VOOs)2/ 

10.1.2.  Net contents 

The net contents shall be declared by volume in the metric system ("Système International" units). 

10.1.3.  Name and address 

The name and address of the manufacturer, packer, distributor, importer, exporter or seller shall be declared. 

________________ 
1/ 

This product may only be sold direct to the consumer if permitted in the country of retail sale. 

2/ 

The country of retail sale may require a more specific designation. 

 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 12 

10.1.4.  Country of origin 

The name of the country of origin shall be declared. When the product undergoes substantial processing in 
a second country, the country in which such processing is carried out shall be considered as the country of 
origin for labelling purposes. 

10.1.5.  Geographical indications and designations of origin 

10.1.5.1. Geographical indications 

The labels of virgin olive oils may state their geographical indication (country, region or locality) when they 
have  been  empowered  to  do  so  by  their  country  of  origin  and  when  such  virgin  olive  oils  have  been 
produced, packed and originate exclusively in the country, region or locality mentioned. 

10.1.5.2. Designations of origin 

The labels of extra virgin olive oils may indicate their designation of origin (country, region or locality) 
when  they  have  been  awarded  such  a  designation,  in  accordance  with  the  terms  provided  under  the 
regulations of their country of origin and when such extra virgin olive oil has been produced, packed and 
originates exclusively in the country, region or locality mentioned. 

10.1.6.  Lot identification 

Each  container  shall  be  embossed  or  otherwise  permanently  marked  in  code  or  in  clear  to  identify  the 
producing factory and the lot. 

10.1.7.  Date marking and storage conditions 

10.1.7.1. Date of minimum durability 

In  the  case  of  pre-packaged  products  intended  for  the  end  consumer,  the  date  of  minimum  durability 
(preceded by the words "best before end") shall be declared by the month and year in uncoded numerical 
sequence. The month may be indicated by letters in those countries where such use will not confuse the 
consumer; if the shelf life of the product is valid to December, the expression "end (stated year)" may be 
used as an alternative. 

10.1.7.2. Storage instructions 

Any special conditions for storage shall be declared on the label if the validity of the date of minimum 
durability depends thereon. 

 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 13 

10.2.  On forwarding packs of oils intended for human consumption 

In addition to the details noted under section 10.1., the following inscription shall appear: 
- number and type of containers held in pack. 

10.3.  On containers allowing the transportation in bulk of olive oils and olive pomace oils 

The labelling on each container shall include: 

10.3.1.  Name of the product 

The name shall indicate the specific designation of the product contained, complying in every way with the 
provisions of this standard. 

10.3.2.  Net contents 

The net contents shall be declared by weight or volume in the metric system ("Système International" units). 

10.3.3.  Name and address 

The name and address of the manufacturer, distributor or exporter shall be declared. 

10.3.4.  Country of origin 

The name of the exporting country shall be declared. 

11. 

METHODS OF ANALYSIS AND SAMPLING 

The methods of analysis and sampling given below are international referee methods. The latest version of 
these methods should be used. 

11.1. 

Sampling 

According to ISO 5555, "Animal and vegetable fats and oils - Sampling". 

11.2. 

Preparation of the test sample 

According to ISO 661, "Animal and vegetable fats and oils - Preparation of the test sample". 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 14 

11.3. 

Determination of the fatty acid composition and trans fatty acid content 

According to COI/T.20/Doc. No 33/Rev.2, “Determination of the fatty acid composition in olive and olive-
pomace oils by gas chromatography” 

11.4.  Determination of the sterol composition and content and alcoholic compounds 

According to COI/T.20/Doc. No 26/Rev. 5, “Determination of the composition and content of sterols, 
triterpenic dialcohols and aliphatic alcohols by capillary column gas chromatography”. 

11.5.  Determination of the difference between the actual and theoretical ECN 42 triacylglycerol content 

According to COI/T.20/Doc. No 20/Rev. 4, "Determination of the difference between actual and theoretical 
content of triacylglycerols with ECN 42", or AOCS 5b-89. 

11.6.  Determination of the stigmastadiene content 

According  to  COI/T.20/Doc.  No  11/Rev.4,  "Determination  of  stigmastadienes  in  vegetable  oils",  or 
COI/T.20/Doc.  no.  16/Rev.2,  "Determination  of  sterenes  in  refined  vegetable  oils",  or  ISO  15788-1  or 
AOCS Cd 26-96. 

11.7.  Determination of the content of 2-glyceryl monopalmitate 

According to COI/T.20/Doc. No 23/Rev.1, "Determination of the percentage of 2-glyceryl monopalmitate" 
or to ISO 12872. 

11.8.  Determination of the unsaponifiable matter 

According  to  ISO  3596,  “Determination  of  the  unsaponifiable  matter  –  Method  using  diethyl  ether 
extraction”, or AOCS Ca 6b-53 or ISO 18609. 

The results should be expressed in g/unsaponifiable matter per kg/oil. 

11.9.  Determination of the organoleptic characteristics 

According to COI/T.20/Doc. No 15/Rev. 10 11, "Organoleptic assessment of virgin olive oil". 

11.10.  Determination of the free acidity 

According to COI/T.20/Doc. No 34/Rev.1, “Determination of free fatty acids, cold method”. 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 15 

11.11.  Determination of the peroxide value 

According 
ISO 3960, or AOCS Cd 8b-90. 

to  COI/T.20/Doc.  No  35/Rev.1, 

“Determination  of 

the  peroxide  value”, 

11.12.  Determination of the absorbency in ultra-violet 

According to COI/T.20/Doc. No 19/Rev.5, "Spectrophotometric investigation in the ultraviolet", or ISO 
3656 or AOCS Ch 5-91. 

11.13.  Determination of the moisture and volatile matter 

According to ISO 662, "Determination of the moisture and volatile matter". 

11.14.  Determination of the insoluble impurities in light petroleum 

According to ISO 663, "Determination of the insoluble impurities". 

11.15.  Determination of the flash point 

According to the FOSFA International method. 

11.16.  Detection of trace metals 

According  to  ISO  8294,  "Determination  of  copper,  iron  and  nickel  by  direct  graphite  furnace  atomic 
absorption spectrometry". 

11.17.  Determination of the alpha-tocopherol 

According  to  ISO  9936, “Determination  of  tocopherols  and tocotrienols  contents  –  Method  using  high-
performance liquid chromatography”. 

11.18.  Determination of traces of heavy metals 

-  Determination of lead: according to ISO 12193 or AOCS Ca 18c-91 or AOAC 994.02. 

- Determination of arsenic: according to AOAC 952.13 or AOAC 942.17 or AOAC 985.16. 

11.19.  Detection of traces of halogenated solvents 

According  to  COI/T.20/Doc.  No  8,  “Determination  of  tetrachloroethylene  in  olive  oils  by  gas-liquid 
chromatography”. 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 20 
page 16 

11.20.  Determination of the content of waxes and alkyl esters 

[According to COI/T.20/Doc. No 28/Rev.3 Corr 1, “Determination of the content of waxes, fatty acid methyl 
esters and fatty acid ethyl esters by capillary gas chromatography“. 

11.21.  Determination of biophenols 

According to COI/T.20/Doc. No 29/Rev.2,  “Document to declare the use of IOC methods for  phenol 
determination” 

11.22.  Determination of the coherence of TAG composition with the fatty acid composition: Screening method 

(not legal in nature and cannot downgrade an oil) 

According  to  COI/T.20/Doc.  No  25/Rev.2,  “Method  for  the  evaluation  of  the  coherence  of  TAG 
composition with the fatty acid composition”. 

11.23.  Determination of the methanol and ethanol content in virgin olive oils 

According to COI/T.20/Doc. No 36, “Method of determination of ethanol and methanol content on virgin olive 
oils”.