SIST EN ISO 12872:2014
(Main)Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010)
Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010)
EN ISO 12872 specifies a procedure for the determination of the content, as a percentage mass fraction, of 2-glyceryl monopalmitate in olive oils and olive-pomace oils that are liquid at ambient temperature (20 °C).
Olivenöle und Oliventresteröle - Verfahren zur Bestimmung des Prozentanteiles von 2-Glyceryl Monopalmitat in Olivenölen (ISO 12872:2010)
Diese Internationale Norm beschreibt ein Verfahren zur Bestimmung des Gehalts, als Massenanteil in Prozent, an 2-Glycerylmonopalmitat in Olivenölen und Oliventresterölen, die bei Umgebungstemperatur (20 °C) flüssig sind.
ANMERKUNG Diese Internationale Norm basiert auf COI/T.20/Doc. 23:2006 [6].
Huiles d'olive et huiles de grignons d'olive - Détermination de la teneur en 2-glycéryl monopalmitate (ISO 12872:2010)
L'ISO 12872:2010 spécifie une méthode pour la détermination de la teneur, sous forme de fraction massique en pourcentage, en 2-glycéryl monopalmitate des huiles d'olive et des huiles de grignons d'olive liquides à température ambiante (20 °C).
Oljčna olja in olja iz oljčnih tropin - Določevanje vsebnosti 2-glyceryl monopalmitata (ISO 12872:2010)
Standard EN ISO 12872 navaja postopek za določevanje vsebnosti 2-gliceril monopalmitata kot odstotek masnega deleža v olivnih oljih in oljih iz oljčnih tropin, ki so pri temperaturi okolja (20° C) v tekočem stanju.
General Information
Relations
Standards Content (Sample)
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.O\FHU\OOlivenöle und Oliventresteröle - Verfahren zur Bestimmung des Prozentanteiles von 2-Glyceryl Monopalmitat in Olivenölen (ISO 12872:2010)Huiles d'olive et huiles de grignons d'olive - Détermination de la teneur en 2-glycéryl monopalmitate (ISO 12872:2010)Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010)67.200.10Animal and vegetable fats and oilsICS:Ta slovenski standard je istoveten z:EN ISO 12872:2014SIST EN ISO 12872:2014en01-julij-2014SIST EN ISO 12872:2014SLOVENSKI
STANDARD
SIST EN ISO 12872:2014
EUROPEAN STANDARD NORME EUROPÉENNE EUROPÄISCHE NORM
EN ISO 12872
April 2014 ICS 67.200.10 English Version
Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content (ISO 12872:2010)
Huiles d'olive et huiles de grignons d'olive - Détermination de la teneur en 2-glycéryl monopalmitate (ISO 12872:2010) Olivenöle und Oliventresteröle - Verfahren zur Bestimmung des Prozentanteiles von 2-Glyceryl Monopalmitat in Olivenölen (ISO 12872:2010) This European Standard was approved by CEN on 11 April 2014.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre:
Avenue Marnix 17,
B-1000 Brussels © 2014 CEN All rights of exploitation in any form and by any means reserved worldwide for CEN national Members. Ref. No. EN ISO 12872:2014 ESIST EN ISO 12872:2014
EN ISO 12872:2014 (E) 2 Contents Page Foreword .3 SIST EN ISO 12872:2014
EN ISO 12872:2014 (E) 3 Foreword The text of ISO 12872:2010 has been prepared by Technical Committee ISO/TC 34 “Food products” of the International Organization for Standardization (ISO) and has been taken over as EN ISO 12872:2014 by Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products - Methods of sampling and analysis” the secretariat of which is held by AFNOR. This European Standard shall be given the status of a national standard, either by publication of an identical text or by endorsement, at the latest by October 2014, and conflicting national standards shall be withdrawn at the latest by October 2014. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights. According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom. Endorsement notice The text of ISO 12872:2010 has been approved by CEN as EN ISO 12872:2014 without any modification.
SIST EN ISO 12872:2014
SIST EN ISO 12872:2014
Reference numberISO 12872:2010(E)© ISO 2010
INTERNATIONAL STANDARD ISO12872First edition2010-05-01Olive oils and olive-pomace oils — Determination of the 2-glyceryl monopalmitate content Huiles d'olive et huiles de grignons d'olive — Détermination de la teneur en 2-glycéryl monopalmitate
SIST EN ISO 12872:2014
ISO 12872:2010(E) PDF disclaimer This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat accepts no liability in this area. Adobe is a trademark of Adobe Systems Incorporated. Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
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©
ISO 2010 All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO's member body in the country of the requester. ISO copyright office Case postale 56 • CH-1211 Geneva 20 Tel.
+ 41 22 749 01 11 Fax
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copyright@iso.org Web
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ii © ISO 2010 – All rights reserved
SIST EN ISO 12872:2014
ISO 12872:2010(E) © ISO 2010 – All rights reserved iii Contents Page Foreword.iv Introduction.v 1 Scope.1 2 Normative references.1 3 Terms and definitions.1 4 Principle.1 5 Reagents.1 6 Apparatus.2 7 Sampling.3 8 Preparation of the test sample.3 9 Procedure.4 9.1 Preparatory steps.4 9.2 Column chromatography.4 9.3 Hydrolysis with pancreatic lipase.4 9.4 Preparation of the silylated derivatives and gas chromatography.5 9.5 Gas chromatography.5 10 Expression of results.5 11 Precision.6 11.1 Interlaboratory test.6 11.2 Repeatability.6 11.3 Reproducibility.6 12 Test report.6 Annex A (informative)
Chromatograms.7 Annex B (informative)
Results of an interlaboratory test.10 Annex C (informative)
Preparation and activity of the lipase.11 Bibliography.13
SIST EN ISO 12872:2014
ISO 12872:2010(E) iv © ISO 2010 – All rights reserved Foreword ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organizations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization. International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2. The main task of technical committees is to prepare International Standards. Draft International Standards adopted by the technical committees are circulated to the member bodies for voting. Publication as an International Standard requires approval by at least 75 % of the member bodies casting a vote. Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. ISO 12872 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal and vegetable fats and oils. SIST EN ISO 12872:2014
ISO 12872:2010(E) © ISO 2010 – All rights reserved v Introduction As part of the Trade standard applying to olive oils and olive-pomace oils, the International Olive Council (IOC) published COI/T.20/Doc. 23:2006[6]. COI/T.20/Doc. 23 was applicable to olive and olive-pomace oils and was used to distinguish between lampante virgin olive oils and crude olive-pomace oils. Olive pomace is the residual paste which still contains a variable amount of water and oil after pressing or centrifuging. In 2008, the IOC submitted the document to ISO/TC 34/SC 11 for adoption as an International Standard.
SIST EN ISO 12872:2014
SIST EN ISO 12872:2014
INTERNATIONAL STANDARD ISO 12872:2010(E) © ISO 2010 – All rights reserved 1 Olive oils and olive-pomace oils — Determination of the
2-glyceryl monopalmitate content 1 Scope This International Standard specifies a procedure for the determination of the content, as a percentage mass fraction, of 2-glyceryl monopalmitate in olive oils and olive-pomace oils that are liquid at ambient temperature (20 °C). NOTE This International Standard is based on COI/T.20/Doc. 23:2006[6]. 2 Normative references The following referenced documents are indispensable for the application of this document. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including any amendments) applies. ISO 661, Animal and vegetable fats and oils — Preparation of test sample 3 Terms and definitions For the purposes of this document, the following terms and definitions apply. 3.1 2-glyceryl monopalmitate content mass fraction of 2-glyceryl monopalmitate in the monoacylglycerol fraction, determined according to the method specified in this International Standard NOTE The 2-glyceryl monopalmitate content is expressed as a percentage. 4 Principle The oil, after suitable preparation, is subjected to the action of pancreatic lipase. A partial hydrolysis takes place that is specific for positions 1 and 3 of the triacylglycerol molecule so that 2-monoacylglycerols are obtained as reaction products. The percentage of 2-glyceryl monopalmitate in the monoacylglycerol fraction is determined, after silylation, by capillary gas chromatography. 5 Reagents WARNING — Comply with any local regulations which specify the handling of hazardous substances. Technical, organizational and personal safety measures shall be followed. During the analysis, unless otherwise stated, use only reagents of recognized analytical grade, and distilled or demineralized water or water of equivalent purity. SIST EN ISO 12872:2014
ISO 12872:2010(E) 2 © ISO 2010 – All rights reserved 5.1 Silica gel, with a particle size of 0,063 mm to 0,200 mm (70/280 mesh), prepared as follows: put the silica gel into a porcelain cup, dry in an oven at 160 °C for 4 h, then cool at ambient temperature in a desiccator. Add a volume of water equivalent to 5 % of the mass of the silica gel as follows: weigh 152 g of silica gel into a 500 ml Erlenmeyer flask, add 8 g of water, stopper and homogenize carefully. Leave to settle for at least 12 h before using. 5.2 n-Hexane, chromatography grade. 5.3 Isopropanol. 5.4 Isopropanol-water mixture, volume fractions 50 ml/100 ml. 5.5 Pancreatic lipase, activity between 2,0 and 10 lipase units per milligram (see Annex C). 5.6 Buffer solution of tris(hydroxymethyl)aminomethane: prepare an aqueous solution (1 mol/l) with pH 8 and mix with concentrated HCl, volume fractions 50 ml/100 ml. 5.7 Sodium cholate, special enzyme grade, aqueous solution, mass fraction 0,1 g/100 g. Use this solution within 15 days of preparation. 5.8 Calcium chloride, aqueous solution, mass fraction 22 g/100 g. 5.9 Diethyl ether, chromatography grade. 5.10 Elution solvent: mixture of n-hexane-diethyl ether, volume fraction of n-hexane 87 ml/100 ml and of diethyl ether 13 ml/100 ml. 5.11 Sodium hydroxide, aqueous solution, mass fraction 12 g/100 g. 5.12 Phenolphthalein, ethanolic solution, mass concentration 1 g/100 ml. 5.13 Carrier gas: hydrogen or helium, gas chromatography grade. 5.14 Auxiliary gases: hydrogen, free from moisture and organic substances, and synthetic air, gas chromatography grade. 5.15 Silylation reagent: mixture of pyridine, hexamethyldisilazane (HMDS) and trimethylchlorosilane (TMCS); volume fractions: 9 ml/13 ml, 3 ml/13 ml, and 1 ml/13 ml, respectively. 5.16 Reference samples: pure monoacylglycerols and mixtures of monoacylglycerols with a known composition similar to that of the sample. 6 Apparatus Usual laboratory equipment and in particular the following. 6.
...
SLOVENSKI STANDARD
kSIST FprEN ISO 12872:2014
01-januar-2014
2OMþQDROMDLQROMDL]ROMþQLKWURSLQ'RORþHYDQMHJO\FHU\OPRQRSDOPLWDWD ,62
Olive oils and olive-pomace oils - Determination of the 2-glyceryl monopalmitate content
(ISO 12872:2010)
Olivenöle und Oliventresteröle - Verfahren zur Bestimmung des Prozentanteiles von 2-
Glyceryl Monopalmitat in Olivenölen (ISO 12872:2010)
Huiles d'olive et huiles de grignons d'olive - Détermination de la teneur en 2-glycéryl
monopalmitate (ISO 12872:2010)
Ta slovenski standard je istoveten z: FprEN ISO 12872
ICS:
67.200.10 5DVWOLQVNHLQåLYDOVNH Animal and vegetable fats
PDãþREHLQROMD and oils
kSIST FprEN ISO 12872:2014 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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kSIST FprEN ISO 12872:2014
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kSIST FprEN ISO 12872:2014
EUROPEAN STANDARD
FINAL DRAFT
FprEN ISO 12872
NORME EUROPÉENNE
EUROPÄISCHE NORM
October 2013
ICS 67.200.10
English Version
Olive oils and olive-pomace oils - Determination of the 2-glyceryl
monopalmitate content (ISO 12872:2010)
Huiles d'olive et huiles de grignons d'olive - Détermination Olivenöle und Oliventresteröle - Verfahren zur Bestimmung
de la teneur en 2-glycéryl monopalmitate (ISO 12872:2010) des Prozentanteiles von 2-Glyceryl Monopalmitat in
Olivenölen (ISO 12872:2010)
This draft European Standard is submitted to CEN members for unique acceptance procedure. It has been drawn up by the Technical
Committee CEN/TC 307.
If this draft becomes a European Standard, CEN members are bound to comply with the CEN/CENELEC Internal Regulations which
stipulate the conditions for giving this European Standard the status of a national standard without any alteration.
This draft European Standard was established by CEN in three official versions (English, French, German). A version in any other language
made by translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are aware and to
provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without notice and
shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2013 CEN All rights of exploitation in any form and by any means reserved Ref. No. FprEN ISO 12872:2013: E
worldwide for CEN national Members.
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kSIST FprEN ISO 12872:2014
FprEN ISO 12872:2013 (E)
Contents Page
Foreword .3
2
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kSIST FprEN ISO 12872:2014
FprEN ISO 12872:2013 (E)
Foreword
The text of ISO 12872:2010 has been prepared by Technical Committee ISO/TC 34 “Food products” of the
International Organization for Standardization (ISO) and has been taken over as FprEN ISO 12872:2013 by
Technical Committee CEN/TC 307 “Oilseeds, vegetable and animal fats and oils and their by-products -
Methods of sampling and analysis” the secretariat of which is held by AFNOR.
This document is currently submitted to the Unique Acceptance Procedure.
Endorsement notice
The text of ISO 12872:2010 has been approved by CEN as FprEN ISO 12872:2013 without any modification.
3
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kSIST FprEN ISO 12872:2014
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kSIST FprEN ISO 12872:2014
INTERNATIONAL ISO
STANDARD 12872
First edition
2010-05-01
Olive oils and olive-pomace oils —
Determination of the 2-glyceryl
monopalmitate content
Huiles d'olive et huiles de grignons d'olive — Détermination de la teneur
en 2-glycéryl monopalmitate
Reference number
ISO 12872:2010(E)
©
ISO 2010
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kSIST FprEN ISO 12872:2014
ISO 12872:2010(E)
PDF disclaimer
This PDF file may contain embedded typefaces. In accordance with Adobe's licensing policy, this file may be printed or viewed but
shall not be edited unless the typefaces which are embedded are licensed to and installed on the computer performing the editing. In
downloading this file, parties accept therein the responsibility of not infringing Adobe's licensing policy. The ISO Central Secretariat
accepts no liability in this area.
Adobe is a trademark of Adobe Systems Incorporated.
Details of the software products used to create this PDF file can be found in the General Info relative to the file; the PDF-creation
parameters were optimized for printing. Every care has been taken to ensure that the file is suitable for use by ISO member bodies. In
the unlikely event that a problem relating to it is found, please inform the Central Secretariat at the address given below.
COPYRIGHT PROTECTED DOCUMENT
© ISO 2010
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or
ISO's member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2010 – All rights reserved
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kSIST FprEN ISO 12872:2014
ISO 12872:2010(E)
Contents Page
Foreword .iv
Introduction.v
1 Scope.1
2 Normative references.1
3 Terms and definitions .1
4 Principle.1
5 Reagents.1
6 Apparatus.2
7 Sampling.3
8 Preparation of the test sample.3
9 Procedure.4
9.1 Preparatory steps .4
9.2 Column chromatography.4
9.3 Hydrolysis with pancreatic lipase.4
9.4 Preparation of the silylated derivatives and gas chromatography .5
9.5 Gas chromatography .5
10 Expression of results.5
11 Precision.6
11.1 Interlaboratory test.6
11.2 Repeatability .6
11.3 Reproducibility .6
12 Test report.6
Annex A (informative) Chromatograms .7
Annex B (informative) Results of an interlaboratory test .10
Annex C (informative) Preparation and activity of the lipase.11
Bibliography.13
© ISO 2010 – All rights reserved iii
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kSIST FprEN ISO 12872:2014
ISO 12872:2010(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards bodies
(ISO member bodies). The work of preparing International Standards is normally carried out through ISO
technical committees. Each member body interested in a subject for which a technical committee has been
established has the right to be represented on that committee. International organizations, governmental and
non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the
International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. ISO shall not be held responsible for identifying any or all such patent rights.
ISO 12872 was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
iv © ISO 2010 – All rights reserved
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kSIST FprEN ISO 12872:2014
ISO 12872:2010(E)
Introduction
As part of the Trade standard applying to olive oils and olive-pomace oils, the International Olive Council
[6]
(IOC) published COI/T.20/Doc. 23:2006 . COI/T.20/Doc. 23 was applicable to olive and olive-pomace oils
and was used to distinguish between lampante virgin olive oils and crude olive-pomace oils. Olive pomace is
the residual paste which still contains a variable amount of water and oil after pressing or centrifuging.
In 2008, the IOC submitted the document to ISO/TC 34/SC 11 for adoption as an International Standard.
© ISO 2010 – All rights reserved v
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kSIST FprEN ISO 12872:2014
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kSIST FprEN ISO 12872:2014
INTERNATIONAL STANDARD ISO 12872:2010(E)
Olive oils and olive-pomace oils — Determination of the
2-glyceryl monopalmitate content
1 Scope
This International Standard specifies a procedure for the determination of the content, as a percentage mass
fraction, of 2-glyceryl monopalmitate in olive oils and olive-pomace oils that are liquid at ambient temperature
(20 °C).
[6]
NOTE This International Standard is based on COI/T.20/Doc. 23:2006 .
2 Normative references
The following referenced documents are indispensable for the application of this document. For dated
references, only the edition cited applies. For undated references, the latest edition of the referenced
document (including any amendments) applies.
ISO 661, Animal and vegetable fats and oils — Preparation of test sample
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
3.1
2-glyceryl monopalmitate content
mass fraction of 2-glyceryl monopalmitate in the monoacylglycerol fraction, determined according to the
method specified in this International Standard
NOTE The 2-glyceryl monopalmitate content is expressed as a percentage.
4 Principle
The oil, after suitable preparation, is subjected to the action of pancreatic lipase. A partial hydrolysis takes
place that is specific for positions 1 and 3 of the triacylglycerol molecule so that 2-monoacylglycerols are
obtained as reaction products. The percentage of 2-glyceryl monopalmitate in the monoacylglycerol fraction is
determined, after silylation, by capillary gas chromatography.
5 Reagents
WARNING — Comply with any local regulations which specify the handling of hazardous substances.
Technical, organizational and personal safety measures shall be followed.
During the analysis, unless otherwise stated, use only reagents of recognized analytical grade, and distilled or
demineralized water or water of equivalent purity.
© ISO 2010 – All rights reserved 1
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kSIST FprEN ISO 12872:2014
ISO 12872:2010(E)
5.1 Silica gel, with a particle size of 0,063 mm to 0,200 mm (70/280 mesh), prepared as follows: put the
silica gel into a porcelain cup, dry in an oven at 160 °C for 4 h, then cool at ambient temperature in a
desiccator. Add a volume of water equivalent to 5 % of the mass of the silica gel as follows: weigh 152 g of
silica gel into a 500 ml Erlenmeyer flask, add 8 g of water, stopper and homogenize carefully. Leave to settle
for at least 12 h before using.
5.2 n-Hexane, chromatography grade.
5.3 Isopropanol.
5.4 Isopropanol-water mixture, volume fractions 50 ml/100 ml.
5.5 Pancreatic lipase, activity between 2,0 and 10 lipase units per milligram (see Annex C).
5.6 Buffer solution of tris(hydroxymethyl)aminomethane: prepare an aqueous solution (1 mol/l) with
pH 8 and mix with concentrated HCl, volume fractions 50 ml/100 ml.
5.7 Sodium cholate, special enzyme grade, aqueous solution, mass fraction 0,1 g/100 g.
Use this solution within 15 days of preparation.
5.8 Calcium chloride, aqueous solution, mass fraction 22 g/100 g.
5.9 Diethyl ether, chromatography grade.
5.10 Elution solvent: mixture of n-hexane-diethyl ether, volume fraction of n-hexane 87 ml/100 ml and of
diethyl ether 13 ml/100 ml.
5.11 Sodium hydroxide, aqueous solution, mass fraction 12 g/100 g.
5.12 Phenolphthalein, ethanolic solution, mass concentration 1 g/100 ml.
5.13 Carrier gas: hydrogen or helium, gas chromatography grade.
5.14 Auxiliary gases: hydrogen, free from moisture and organic substances, and synthetic air, gas
chromatography grade.
5.15 Silylation reagent: mixture of pyridine, hexamethyldisilazane (HMDS) and trimethylchlorosilane
(TMCS); volume fractions: 9 ml/13 ml, 3 ml/13 ml, and 1 ml/13 ml, respectively.
5.16 Reference samples: pure monoacylglycerols and mixtures of monoacylglycerols with a known
composition similar to that of the sample.
6 Apparatus
Usual laboratory equipment and in particular the following.
6.1 Erlenmeyer flasks, of capacity 25 ml.
6.2 Beakers, of capacities 100 ml, 250 ml, and 300 ml.
6.3 Glass chromatography column, 21 mm to 23 mm internal diameter, 400 mm in length, with septum
and
...
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