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 the trade standard applying to olive oils and olive-pomace oils
Date: 2022-09-28
Institutions: Directorate-General for Agriculture and Rural Development, European Commission
Subjects: agreement (EU), commodity agreement, harmonisation of standards, joint body (EU), olive, olive oil, trade policy

EN 

  EN 

   EUROPEAN COMMISSION  Brussels, 28.9.2022  SWD(2022) 309 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 the trade standard applying to olive oils and olive-pomace oils {COM(2022) 487 final}  
 
 
Draft Decision NO DEC-III.X/116-VI/2022 
revising the trade standard applying to olive oils and 
olive-pomace oils 

EN 

  EN 

 
 
 
 
DRAFT DECISION Nº DEC-III.X/116-VI /2022 

REVISING THE TRADE STANDARD APPLYING TO OLIVE OILS AND OLIVE 
POMACE OILS 

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 11th meeting during the 116th 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 10 and 11 March 2022, on: 

- The removal of Annex 1 and the simplification of the delta 7 stigmastenol decision trees. 

- The inclusion of revision 3 of the method on the determination of waxes and ethyl esters. 

Considering the work carried out by the expert chemists as part of the collaborative trials on the 
validation of the method to determine waxes and  ethyl esters by the revision COI/T.20/Doc. No. 
28/Rev. 3 ‘Determination of the content of waxes and fatty acid ethyl esters by capillary column gas 
chromatography’; 

DECIDES 

1. 

To adopt the Trade Standard for Olive Oils and Olive Pomace Oils COI/T.15/NC 
No.  3/Rev.  19,  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. 18 of June 
2022; 

EN 

  EN 

 
 
 
 
 
 
 
 
 
2. 

3. 

Members shall, in accordance with their respective legislation, take all appropriate 
measures to implement the adopted Standard and shall communicate these measures 
to the Executive Secretariat as soon as they are taken; 

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), November 2022 

Mr Kaled Musa Al Henefat 

Chair of the International Olive Council 

EN 

  EN 

 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 
November 2022 

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. 

EN 

  EN 

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

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

3.2. 

Trans fatty acid content (% trans fatty acids) 

-  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 
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 
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 

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) 

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

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 
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 
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 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 
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: 

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

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; 

8.1. 
oils; 

8.2. 

8.3. 

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. 

EN 

  EN 

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

EN 

  EN 

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

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 
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". 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.15/NC No 3/Rev. 19 
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, "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”. 

EN 

  EN 

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

11.11.  Determination of the peroxide value 

According  to  COI/T.20/Doc.  No  35/Rev.1,  “Determination  of  the  peroxide  value”, 
ISO 3960, or AOCS Cd 8b-90. 

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”. 

EN 

  EN 

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

11.20.  Determination of the content of waxes and alkyl esters 

[According to COI/T.20/Doc. No 28/Rev.3, “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”. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
DECISION Nº DEC-III-X/116-VI/202 concerning the 
method for the determination of the content of waxes 
and fatty acid ethyl esters by capillary gas 
chromatography 

EN 

  EN 

 
 
 
DECISION Nº DEC-III-X/116-VI/2022 

CONCERNING THE METHOD FOR THE DETERMINATION OF THE CONTENT OF 
WAXES AND 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 the recommendation made by the Chemistry and Standardisation Committee at its 
11th meeting during the 116th session of the Council of Members; 

Considering the chemistry experts studied the application of method COI/T.20/Doc. No 28 and 
its fusion with method COI/T.20/Doc. No 31; 

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 10 and 11 March 2022; 

DECIDES 

To adopt revision 3 of the document COI/T.20/Doc. No 28/Rev. 3 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. 

This revised method COI/T.20/Doc. No 28/Rev. 3 shall replace the methods COI/T.20/Doc. No 
28/Rev. 2 and COI/T.20/Doc. No 31 and will be listed in the IOC trade standard. 

Madrid (Spain), XX November 2022 

Mr Kaled Musa Al Henefat 
Chair of the International Olive Council 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
November 2022 

ENGLISH 

Original: ENGLISH 

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. 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. 

This document presents two methods that can be used for official control. Method A (15 g of silica) 
is the reference method for counter-assessment. 

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. 

APPARATUS 

3.1. 

Test tube, 10 ml. 

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

suitable stopcock. 

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

on-column injection comprising: 

3.3.1.  Thermostat-controlled oven with temperature programming. 

3.3.2.  Cold injector for direct on-column injection 

3.3.3.  Flame ionisation detector and converter-amplifier. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 2 

3.3.4.  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.  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.  Microsyringe, 10 l, with hardened needle, for direct on-column injection 

3.5. 

3.6. 

Electric shaker. 

Rotary evaporator. 

3.7.  Muffle oven. 

3.8. 

3.9. 

4. 

4.1. 

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

Usual laboratory glassware. 

REAGENTS 

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 not be necessary. 

4.2. 

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 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. 

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 

________________________________ 
Note 1:  Computerised systems may also be used where the gas chromatography data are entered through a PC. 
Note 2:  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 

EN 

  EN 

 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 3 

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. 

n-heptane, chromatography grade, or iso-octane. 

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. 

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

Standard solution of methyl heptadecanoate at 0.005% (m/V) in heptane (internal standard for 
methyl and ethyl esters). 

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

4.5. 

4.6. 

4.7. 

4.8. 

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. 

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. 

______________________ 

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

EN 

  EN 

 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 4 

4.9. 

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. 

PROCEDURE 

5.1. 

Preparation of the chromatography column 

Suspend 15 g of silica gel (4.1) in n-hexane (4.2) and introduce into the column (3.2). Allow to 
settle  spontaneously.  Complete  settling  with  the  aid  of  an  electric  shaker  (3.5)  to  make  the 
chromatographic bed more homogeneous. Percolate 20 ml of n-hexane to remove any impurities. 
Weigh exactly 500 mg of the sample into the 10-ml test tube (3.1), using the analytical balance 
(3.8), and add a suitable amount of internal standard (4.5) 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 (4.6). Transfer the prepared sample to the chromatography 
column with the aid of two 2-ml portions of n-hexane (4.2). 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) of n-hexane/ethyl ether (99:1) and collect 150 ml at a 
flow of about 15 drops every 10 seconds. (This fraction contains ethyl esters and waxes). 

Evaporate the resultant fractions in a rotary evaporator (3.6) 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.  Gas chromatography analysis 

5.2.1.  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.). 

EN 

  EN 

 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 5 

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 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. 

_____________________ 

________________________________ 
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. 

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. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 6 

5.2.2.  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 

20°C/min 

5°C/min 

    20°C/min 

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

-  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). 
- 

Instrument sensitivity: suitable for fulfilling the above conditions. 

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. 

5.3. 

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. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
  
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 7 

5.4. 

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. Shows a chromatogram of the FAEE and waxes in an extra virgin olive oil using the 
method A (15 gr). 

Figure 2. Shows a chromatogram of the FAEE and waxes in a lampante olive oil using the method 
A (15 gr). 

5.5.  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 = 

As = 

ms = 

m = 

area corresponding to the peak for the individual ester, in computer counts (peak 
no. 11-13, 14-15, 16-17-18 in fig. 2) 

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

mass of the lauryl arachidate internal standard added, in milligrams 

mass of the sample taken for determination, in grams 

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

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 8 

5.6.  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= 

As= 

ms= 

m = 

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

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

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

mass of the sample taken for determination, in grams. 

6. 

EXPRESSION OF RESULTS 

Report the sum of the contents of the different waxes from C42 to C46 in the case of extra 
virgin  and  virgin  olive  oils  and  from  C40  to  C46  in  the  case  of  other  oils  (Note  7)  in 
milligrams per kilograms of fat (ppm). 

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 one decimal place. 

__________________________________ 
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. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 9 

METHOD B (3 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. 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. 

APPARATUS 

3.1. 

Test tube, 10 ml. 

3.2.  Glass column for liquid chromatography, internal diameter 10 mm, length 40 cm, fitted with a 

suitable stopcock. 

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

on-column injection comprising: 

3.3.1.  Thermostat-controlled oven with temperature programming. 

3.3.2.  Cold injector for direct on-column injection  

3.3.3.  Flame ionisation detector and converter-amplifier. 

3.3.4.  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. 

3.4.5.  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 µm. 

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

3.5. 

Electric shaker. 

3.6. 

Rotary evaporator. 

3.7.  Muffle oven. 

3.8. 

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

3.9. 

Usual laboratory glassware. 

EN 

  EN 

 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 10 

REAGENTS 

4.1. 

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 not be necessary. 

4.2. 

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

200  ml  of  n-hexane  are  evaporated  to  dryness,  residue  is  redissolved  in  100  µl  n-heptane  and 
analysed applying the same gas chromatographic conditions. There must be no peak in the elution 
alkyl esters area. (Hexane can be replaced by Isoctane) 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. 

4.3. 

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. 

n-heptane, chromatography grade, or iso-octane. 

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. 

EN 

  EN 

 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 11 

4.5. 

4.6. 

Standard solution of lauryl arachidate (Note 3), at 0.01% (m/V) in heptane (internal standard for 
waxes). 

Solution of methyl heptadecanoate, at 0.002% (m/V) in heptane (internal standard for methyl and 
ethyl esters). 

4.7. 

Sudan 1 (1-phenylazo-2-naphthol) 

4.8. 

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. 

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 purpose. 

______________________________ 
Note 1:  Computerised systems may also be used where the gas chromatography data are entered through a PC. 
Note2:   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 3:  Palmityl palmitate, myristyl stearate or arachidyl laureate may also be used. 

EN 

  EN 

 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 12 

4.9. 

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. 

PROCEDURE 

5.1. 

Preparation of the chromatography column 

Suspend 3 g of silica gel (4.1) in n-hexane (4.2) and introduce into the column (3.2). Allow to settle 
spontaneously.  Complete  settling  with  the  aid  of  an  electric  shaker  (3.5)  to  make  the 
chromatographic bed more homogeneous. Percolate 10 ml of n-hexane to remove any impurities. 
Weigh exactly 100 mg of the sample into the 10-ml test tube (3.1), using the analytical balance 
(3.8), and add a suitable amount of internal standard (4.5) depending on the assumed wax content, 
e.g. add 0.01 mg of lauryl arachidate in the case of extra virgin olive oil, virgin olive oil, refined 
olive  oil  and  olive  oil,  0.025-0.10  mg  in  the  case  of  olive-pomace  oil  and  0.002  mg  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-ml portions of n-hexane (4.2). 
Allow the solvent to flow to 1 mm above the upper level of the absorbent. Percolate 12-15 ml n-
Hexane (*) to further remove hydrocarbons (alkanes and sterenes) 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). 
Evaporate the resultant fractions in a rotary evaporator (3.7) 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. 

(*) 

Not necessary for the determination of waxes only. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 13 

5.2.  Gas chromatography analysis 

5.2.1.  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 (3.3.4), 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. 

____________________________________ 

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 

Note 5: 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. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 14 

5.2.2.  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 

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). 
- 

Instrument sensitivity: suitable for fulfilling the above conditions. 

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. 

5.3. 

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  are  completely  eluted,  depending  on  the  fraction  being 
analysed. 
The base line must always meet the required conditions 

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

scale value. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 15 

5.4. 

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 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 gr). 

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

5.5.  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 and virgin olive oils and from C40 to 
C46 for 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= 

As= 

ms= 

m = 

area corresponding to the peak for the individual ester, in computer counts (peak 
n° 11- 13, 14-15,16-17-18 in fig.2) 

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

mass of the lauryl arachidate internal standard added, in milligrams 

mass of the sample taken for determination, in grams 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 16 

5.6.  Quantitative analysis of the ethyl esters 

With  the  aid  of  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= 

As = 

ms= 

m = 

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

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

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

mass of the sample taken for determination, in grams. 

6. 

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

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 one decimal place. 

Results should be expressed to two decimal places, up to 1 mg/kg above this with one decimal 
place. 

_________________________ 

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. 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 17 

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 gr). 

EN 

  EN 

 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 18 

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

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

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 19 

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

EN 

  EN 

 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 20 

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.  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 
outliers  
mean 
r 

Sr 
RSDr (%) 
R 

Sr 
RSDr (%) 

number of participating laboratories 
number of laboratories with outlying values 
mean of the accepted results 
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%  
repeatability standard deviation  
repeatability coefficient of variation (Sr x 100 / mean) 
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  
reproducibility coefficient of variation (Sr x 100 / mean) 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 21 

M2 

M1 

Ethyl esters (mg/kg) – Method A 15 g of silica 
Sample 
Mean 
n 
outliers 
Sr 
RSDr (%) 
r 
SR 
RSDR (%) 
R 

8.21 
16 
2 
0.330 
4.02 
0.92 
0.915 
11.15 
2.56 

10 
16 
3 
0.574 
5.74 
1.61 
0.759 
7.59 
2.12 

M3 

M4 

36.97 
16 
0 
1.316 
3.56 
3.68 
3.720 
10.06 
10.42 

50.83 
16 
0 
1.934 
3.80 
5.41 
6.736 
13.25 
18.86 

Sample 
Mean 
n 
outliers 
Sr 
RSDr (%) 
r 
SR 
RSDR (%) 
R 

Sample 
Mean 
n 
Outliers 
Sr 
RSDr (%) 
r 
SR 
RSDR (%) 
R 

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 
323.17 
16 
1 
4.346 
1.34 
12.17 
23.649 
7.32 
66.22 

M5 
2350.72 
16 
0 
35.728 
1.52 
100.04 
247.180 
10.52 
692.10 

Ethyl esters (mg/kg) -Method B  (3 g of silica) 
M2 
8.15 
15 
4 
0.225 
2.76 
0.63 
0.991 
12.16 
2.77 

M1 
10.00 
15 
4 
0.570 
5.90 
1.65 
1.681 
14.00 
3.92 

M3 
39.22 
15 
0 
1.234 
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 

EN 

  EN 

 
 
 
 
 
 
 
 
 
 
COI/T.20/Do. No 28/Rev. 3 
Page 22 

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

Sample 
Mean 
n 
Outliers 
Sr 
RSDr (%) 
r 
SR 
RSDR (%) 
R 

M1 
90.45 
15 
2 
1.770 
1.96 
4.96 
15.287 
16.90 
42.80 

M2 
45.53 
15 
1 
2.306 
5.06 
6.46 
7.066 
15.52 
19.79 

M3 
38.81 
15 
2 
1.745 
4.50 
4.89 
8.236 
21.22 
23.06 

M4 
325.47 
15 
0 
7.758 
2.38 
21.72 
23.614 
7.26 
66.12 

M5 
2354.21 
15 
0 
50.548 
2.15 
141.54 
213.990 
9.09 
599.17 

ANNEX D 

References 

ISO 5725-1:1994 
General principles and definitions  

Accuracy (trueness and precision) of measurement methods and results – Part 1: 

ISO 5725-2:1994 
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 
Alternative methods for the determination of the precision of a standard measurement method  

Accuracy (trueness and precision) of measurement methods and results – Part 5: 

ISO 5725-6:1994 
Use in practice of accuracy values 

Accuracy (trueness and precision) of measurement methods and results – Part 6: 

EN 

  EN