Animal and vegetable fats and oils - Determination of phthalates in vegetable oils

This document specifies two methods for the quantitative determination of phthalates in vegetable oils by gas chromatography-mass spectrometry (GC-MS): - Part A for the determination of di-2-ethylhexyl phthalate (DEHP). - Part B for the determination of eight phthalates: dimethyl phthalate (DMP), diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), di-isononyl phthalate (DINP), di-isodecyl phthalate (DIDP). Both methods are applicable for all vegetable oils, including crude, refined and virgin.

Corps gras d’origines animale et végétale — Dosage des phtalates dans les huiles végétales

Le présent document spécifie deux méthodes de quantification des phtalates dans les huiles végétales par chromatographie en phase gazeuse couplée à la spectrométrie de masse (CG-SM): — Partie A pour le dosage du phtalate de di-2-éthylhexyle (DEHP); — Partie B pour le dosage de huit phtalates: phtalate de diméthyle (DMP), phtalate de diéthyle (DEP), phtalate de di-isobutyle (DIBP), phtalate de dibutyle (DBP), phtalate de benzylbutyle (BBP), phtalate de di-2-éthylhexyle (DEHP), phtalate de di-isononyle (DINP), phtalate de di-isodécyle (DIDP). Les deux méthodes sont applicables aux huiles végétales, qu’il s’agisse d’huile brute, raffinée ou vierge.

General Information

Status
Published
Publication Date
30-Jan-2024
Current Stage
6060 - International Standard published
Start Date
31-Jan-2024
Due Date
07-Jul-2024
Completion Date
31-Jan-2024
Ref Project

Overview

ISO/TS 16465:2024 specifies validated laboratory methods for the quantitative determination of phthalates in vegetable oils using gas chromatography–mass spectrometry (GC‑MS). The Technical Specification is published by ISO and is split into two parts:

  • Part A – targeted determination of di‑2‑ethylhexyl phthalate (DEHP).
  • Part B – determination of eight phthalates: DMP, DEP, DIBP, DBP, BBP, DEHP, DINP, DIDP.

Both methods apply to all vegetable oils (crude, refined and virgin) and cover sample preparation, extraction, clean‑up, instrumental analysis, calibration and method performance characteristics.

Key topics and technical requirements

  • Analytical principle: extraction with acetonitrile, dispersive solid‑phase extraction (dispersive‑SPE) clean‑up using PSA (primary secondary amine) and C18 sorbents, concentration of the extract and GC‑MS analysis (SIM mode).
  • Target analytes: DEHP (Part A) and a panel of eight phthalates (Part B): DMP, DEP, DIBP, DBP, BBP, DEHP, DINP, DIDP.
  • Reagents and standards: toluene, acetonitrile, n‑hexane, n‑dodecane, PSA and C18 sorbents; use of a deuterated internal standard (DEHP‑d4) and a defined calibration series.
  • Instrumentation: capillary GC with MS detector (electron ionization at 70 eV), single ion monitoring (SIM), and high‑temperature capillary columns suitable for phthalate analysis.
  • Apparatus and cleanliness: strict glassware cleaning (e.g., 550 °C for 6 h), centrifugation, micro‑syringes and optional automated evaporator for concentration steps.
  • Quality and validation: procedures for calibration curves, quantitative calculation, and documented precision data (interlaboratory tests, repeatability and reproducibility).
  • Sampling and sample handling: analyse aliquots supplied in glass bottles; recommended lid contact materials include PTFE, aluminium or rubber - avoid plastic contact materials.

Applications and users

ISO/TS 16465:2024 supports:

  • Food and edible oil testing laboratories performing compliance and contamination testing for phthalates in vegetable oils.
  • Oil producers and refiners for in‑house quality control and supplier verification.
  • Regulatory authorities and food safety agencies monitoring phthalate residues in edible fats and oils.
  • Research institutions studying phthalate migration, exposure or processing impacts.
  • Contract testing and environmental labs when assessing phthalate contamination from packaging or processing.

Keywords: ISO/TS 16465:2024, phthalates in vegetable oils, DEHP determination, GC‑MS phthalate analysis, dispersive solid‑phase extraction, PSA C18 cleanup.

Related standards

  • ISO 661 - Preparation of test sample for animal and vegetable fats and oils (normative reference).
  • Recommended sampling guidance referenced: ISO 5555 (sampling of vegetable oils).
Technical specification
ISO/TS 16465:2024 - Animal and vegetable fats and oils — Determination of phthalates in vegetable oils Released:31. 01. 2024
English language
25 pages
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Frequently Asked Questions

ISO/TS 16465:2024 is a technical specification published by the International Organization for Standardization (ISO). Its full title is "Animal and vegetable fats and oils - Determination of phthalates in vegetable oils". This standard covers: This document specifies two methods for the quantitative determination of phthalates in vegetable oils by gas chromatography-mass spectrometry (GC-MS): - Part A for the determination of di-2-ethylhexyl phthalate (DEHP). - Part B for the determination of eight phthalates: dimethyl phthalate (DMP), diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), di-isononyl phthalate (DINP), di-isodecyl phthalate (DIDP). Both methods are applicable for all vegetable oils, including crude, refined and virgin.

This document specifies two methods for the quantitative determination of phthalates in vegetable oils by gas chromatography-mass spectrometry (GC-MS): - Part A for the determination of di-2-ethylhexyl phthalate (DEHP). - Part B for the determination of eight phthalates: dimethyl phthalate (DMP), diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-2-ethylhexyl phthalate (DEHP), di-isononyl phthalate (DINP), di-isodecyl phthalate (DIDP). Both methods are applicable for all vegetable oils, including crude, refined and virgin.

ISO/TS 16465:2024 is classified under the following ICS (International Classification for Standards) categories: 67.200.10 - Animal and vegetable fats and oils. The ICS classification helps identify the subject area and facilitates finding related standards.

You can purchase ISO/TS 16465:2024 directly from iTeh Standards. The document is available in PDF format and is delivered instantly after payment. Add the standard to your cart and complete the secure checkout process. iTeh Standards is an authorized distributor of ISO standards.

Standards Content (Sample)


Technical
Specification
ISO/TS 16465
First edition
Animal and vegetable fats and
2024-01
oils — Determination of phthalates
in vegetable oils
Corps gras d’origines animale et végétale — Dosage des phtalates
dans les huiles végétales
Reference number
© ISO 2024
All rights reserved. Unless otherwise specified, or required in the context of its implementation, no part of this publication may
be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on
the internet or an intranet, without prior written permission. Permission can be requested from either ISO at the address below
or ISO’s member body in the country of the requester.
ISO copyright office
CP 401 • Ch. de Blandonnet 8
CH-1214 Vernier, Geneva
Phone: +41 22 749 01 11
Email: copyright@iso.org
Website: www.iso.org
Published in Switzerland
ii
Contents Page
Foreword .iv
1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Part A - Determination of DEHP . 1
4.1 Principle .1
4.2 Reagents .1
4.3 Apparatus .3
4.4 Sampling .4
4.5 Preparation of the test sample .4
4.6 Procedure .4
4.6.1 Phthalates extraction from oil .4
4.6.2 Gas Chromatography .5
4.7 Quantitative determination .6
4.7.1 Calibration curve.6
4.7.2 Quantitative determination of DEHP .6
4.8 Precision of the method.7
4.8.1 Interlaboratory test .7
4.8.2 Repeatability .8
4.8.3 Reproducibility .8
4.9 Test report .8
5 Part B - Determination of eight phthalates. 8
5.1 Principle .8
5.2 Reagents .8
5.3 Apparatus .10
5.4 Sampling .11
5.5 Preparation of the test sample . 12
5.6 Procedure . 12
5.6.1 Phthalates extraction from oil . 12
5.6.2 Gas chromatography .16
5.7 Quantitative determination .17
5.7.1 Calibration curve.17
5.7.2 Quantitative determination of phthalate .18
5.8 Precision of the method.19
5.8.1 Interlaboratory test .19
5.8.2 Repeatability .19
5.8.3 Reproducibility .19
5.9 Test report .19
Annex A (informative) Results of an interlaboratory test — Part A - Determination of DEHP .20
Annex B (informative) Results of an interlaboratory test — Part B - Determination of eight
phthalates . .21
Bibliography .25

iii
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,
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with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.
The procedures used to develop this document and those intended for its further maintenance are described
in the ISO/IEC Directives, Part 1. In particular, the different approval criteria needed for the different types
of ISO document should be noted. This document was drafted in accordance with the editorial rules of the
ISO/IEC Directives, Part 2 (see www.iso.org/directives).
ISO draws attention to the possibility that the implementation of this document may involve the use of (a)
patent(s). ISO takes no position concerning the evidence, validity or applicability of any claimed patent
rights in respect thereof. As of the date of publication of this document, ISO had not received notice of (a)
patent(s) which may be required to implement this document. However, implementers are cautioned that
this may not represent the latest information, which may be obtained from the patent database available at
www.iso.org/patents. ISO shall not be held responsible for identifying any or all such patent rights.
Any trade name used in this document is information given for the convenience of users and does not
constitute an endorsement.
For an explanation of the voluntary nature of standards, the meaning of ISO specific terms and expressions
related to conformity assessment, as well as information about ISO's adherence to the World Trade
Organization (WTO) principles in the Technical Barriers to Trade (TBT), see www.iso.org/iso/foreword.html.
This document was prepared by Technical Committee ISO/TC 34, Food products, Subcommittee SC 11, Animal
and vegetable fats and oils.
Any feedback or questions on this document should be directed to the user’s national standards body. A
complete listing of these bodies can be found at www.iso.org/members.html.

iv
Technical Specification ISO/TS 16465:2024(en)
Animal and vegetable fats and oils — Determination of
phthalates in vegetable oils
1 Scope
This document specifies two methods for the quantitative determination of phthalates in vegetable oils by
gas chromatography-mass spectrometry (GC-MS):
— Part A for the determination of di-2-ethylhexyl phthalate (DEHP).
— Part B for the determination of eight phthalates: dimethyl phthalate (DMP), diethyl phthalate (DEP),
di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-2-ethylhexyl
phthalate (DEHP), di-isononyl phthalate (DINP), di-isodecyl phthalate (DIDP).
Both methods are applicable for all vegetable oils, including crude, refined and virgin.
2 Normative references
The following documents are referred to in the text in such a way that some or all of their content constitutes
requirements 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.
ISO and IEC maintain terminology databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https:// www .iso .org/ obp
— IEC Electropedia: available at https:// www .electropedia .org/
4 Part A - Determination of DEHP
4.1 Principle
Phthalates are first extracted from oil with acetonitrile followed by dispersive-solid-phase extraction
clean-up (dispersive SPE), which is done to remove organic acids, excess water and other components using
a combination of primary secondary amine (PSA) sorbent and C18 bonded phase. Finally, the extract is
concentrated and analysed by gas chromatography-mass spectrometry (GC-MS).
4.2 Reagents
WARNING — Attention is drawn to national regulations that specify the handling of hazardous
substances, and users' obligations thereunder. Technical, organizational and personal safety
measures shall be followed.
Unless otherwise specified, use only reagents of recognized analytical grade.
4.2.1 Toluene, purity 99 % min.

4.2.2 Acetonitrile, trace organic analysis grade, purity 99 % min.
4.2.3 n-Hexane, trace organic analysis grade, purity 99 % min.
1)
4.2.4 Chromabond adsorbent Diamino (PSA), Chromabond adsorbent C18 .
NOTE These powders can be sources of contamination. To remove the contaminants, the powders can be purified
using Soxhlet extraction by extracting twice with 200 ml of n-hexane during a six-hour period.
4.2.5 n-Dodecane, purity 99 % min.
4.2.6 Standards
Table 1 provides the CAS number of phtalates.
Table 1 — CAS number of phthalates
2) ®
Phthalates Synonym CAS Registry Number
di-2-ethylhexyl phthalate DEHP 117-81-7
deuterated di-2-ethylhexyl phthalate DEHP-d4 93951-87-2
4.2.6.1 Solution of DEHP-d4 internal standard, mass concentration ρ = 50 µg/ml.
As an example, in a volumetric flask of 100 ml, weigh to the nearest mg, approximately 50 mg of DEHP-d4
and dilute to 100 ml of toluene and then proceed with a second dilution of this mixture of 10 ml → 100 ml
with toluene.
4.2.6.2 DEHP calibration solutions
As an example, in a volumetric flask of 100 ml, weigh to the nearest mg, approximately 50 mg of DEHP and
dilute to 100 ml of toluene (SM= 500 µg/ml). Then prepare:
— 20 µl of SM and dilute to 10 ml of acetonitrile: SF1 = 1 µg/ml
— 200 µl of SM and dilute to 10 ml of acetonitrile: SF2 = 10 µg/ml
— 2 ml of SM and dilute to 10 ml of acetonitrile: SF3 = 100 µg/ml
Then prepare a calibration range in acetonitrile following Table 2, using glass syringes washed 10 times
with toluene and five times with related solutions.
1) Chromabond sorbent C18 ref 730611 and Chromabond sorbent Diamino (PSA) ref 730611 from Macherey Nagel
(www .mn -net .com) have proved to be free of phthalate contamination. This information is given for the convenience of
users of this document and does not constitute an endorsement by ISO of these products. ®
2) CAS Registry Number is a trademark of the American Chemical Society (ACS). This information is given for the
convenience of users of this document and does not constitute an endorsement by ISO of the product named. Equivalent
products may be used if they can be shown to lead to the same results.

Table 2 — Example of preparation for DEHP calibration
Volume (µl) [Concentration] Solution of
[Concentration]
Volume of
DEHP-d4 DEHP-d4 Total
DEHP
acetoni-
internal volume
C
cal-dph
C trile to be
SF1 SF2 SF3 SM
cal-ph
standard (µl)
µg/ml added (µl)
µg/ml
(µl)
G0 0 - - - - 4 80 920 1 000
G1 0,02 20 - - - 4 80 900 1 000
G2 0,05 50 - - 4 80 870 1 000
G3 0,1 - 10 - - 4 80 910 1 000
G4 0,5 - 50 - - 4 80 870 1 000
G5 1 - - 10 - 4 80 910 1 000
G6 2 - - 20 - 4 80 900 1 000
G7 5 - - - 10 4 80 910 1 000
G8 10 - - - 20 4 80 900 1 000
Depending on the apparatus used, it is not possible to be linear from G0 to G8. In this case, it is necessary to
increase the volume of acetonitrile of recovery in order to be linear over the greatest possible range while
being able to see the level G1.
4.3 Apparatus
Glassware used for the determination shall be thoroughly cleaned at 550 °C during 6 h, such as Pasteur
pipette, vial, conical glass sample vial, 10 ml and 15 ml capacity.
4.3.1 Conical glass sample vials, 10 ml capacity.
4.3.2 Conical glass sample vials, 15 ml capacity.
4.3.3 Glass syringe, 10 µl, 20 µl, 50 µl, 100 µl, 250 µl and 500 µl capacity.
4.3.4 Automatic evaporator, for 10 ml tube (optional), recommended operating conditions: temperature
of the water bath = 40 °C, nitrogen pressure = 5 psi.
4.3.5 Conical glass sample vials, 2 ml capacity.
4.3.6 Gas chromatograph, suitable for use with capillary column, equipped with an injector split-splitless
or equivalent device, a temperature-programmable oven and mass detector with electron ionization source
(ionization energy of 70 eV) and SIM (single ion monitoring) mode.
4.3.7 Data acquisition system, with the possibility of manual integration.
4.3.8 Capillary column, capable of being programmed up to 400 °C ("high temperature" type) for which
the following characteristics are advised: 95 % dimethyl/5 % diphenyl polysiloxane stationary phase,
length of 15 m, internal diameter of 0,25 mm, film thickness of 0,1 µm or length of 30 m, internal diameter of
0,25 mm, 0,25 µm film thickness.
4.3.9 Microsyringe, 5 µl to 10 µl capacity, suitable for split less injection in gas chromatography.
4.3.10 Analytical balance, reading accuracy 0,001 g.
4.3.11 Pasteur pipette, in glass.

4.3.12 Centrifuge, capable of attaining at least 3 000 min−1, suitable for 10 ml tubes.
4.3.13 Pipettes, capable of pipetting volumes up to 10 ml.
4.4 Sampling
A representative sample should have been sent to the laboratory. It should not have been damaged or
changed during transport or storage.
Sampling is not part of the method specified in this document. A recommended sampling method is given in
[1]
ISO 5555.
Analyse only sample aliquots packaged in glass bottles with suitable lids (the sample contact material from
the lid should be, for example, polytetrafluoroethylene (PTFE), aluminium, rubber but not plastic nor soft
seal) to avoid further contamination. Plastic packaging is not suitable.
Due to the presence of phthalates in the environment, the analysis of these compounds requires precautions
throughout the analysis:
— avoid contact with plastic material,
— test the solvents/materials used for phthalates,
— a blank sample should be analysed every four samples,
— glassware that cannot be baked needs to be rinsed with a suitable solvent (acetone),
— avoid contamination from the septum of the injection vials.
4.5 Preparation of the test sample
Prepare the test sample in accordance with ISO 661.
4.6 Procedure
4.6.1 Phthalates extraction from oil
Weigh to the nearest mg, in a conical glass sample vial of 10 ml, 1 g of oil and add 40 µl of DEHP-d4 solution
at 50 µg/ml (4.2.6.1). Shake with a vortex for 15 s.
Add 6 ml of acetonitrile and shake with a vortex for 1 min (or 5 min with an automatic vortex).
Centrifuge the conical glass for 2 min at 3 000 rpm.
Transfer the upper layer in a conical glass of 15 ml containing 400 mg of C18 (4.2.4) and 400 mg of PSA
(4.2.4).
Shake with a vortex for 1 min.
Transfer the upper layer in a conical glass of 10 ml.
Add 50 µl of n-dodecane.
Evaporate acetonitrile at 40 °C to reach 500 µl ± 50 µl.
There remains a droplet of fat at the bottom of the tube from the dodecane.
Shake with a vortex for 15 s (or 2 min with an automatic vortex).
Centrifuge the conical glass for 2 min at 3 000 rpm.
Transfer the solvent layer avoiding the fat droplet in an injection vial and inject into GC-MS.

In order to measure the level of contamination of the laboratory, prepare a blank sample. For blank sample,
add only 40 µl of DEHP-d4 internal standard solution (4.2.6.1) in a conical glass sample vial of 10 ml and
follow the same procedure as for classic sample. (Values of 0,05 mg/kg for DEHP should not be exceeded).
In order to avoid contamination from the septum of the vial (DMP), use a septum with polytetrafluoroethylene
(PTFE), otherwise a small piece of aluminium sheet between the cork and the vial can be used.
NOTE Depending on the apparatus used, the concentration of the extract can be modified and adapted according
to the system.
4.6.2 Gas Chromatography
4.6.2.1 Gas chromatography setup
Install the column (4.3.8) in the gas chromatograph (4.3.6) and check the working conditions by injecting the
solvent, acetonitrile. The baseline should be straight with a small positive drift. If the drift is high, proceed
to condition the column, for a negative drift check the connections of the column.
If the column is used for the first time, it is necessary to condition the column by heating it in the column
oven using a temperature gradient up to 370 °C (depending on the oven temperature chosen for the analysis)
in 4 h, maintain the temperature for 2 h.
4.6.2.2 Working conditions for gas chromatography analysis
The working conditions as shown in Table 3 and Table 4 have proved to be adapted to the analysis.
These conditions can be adjusted in accordance with the characteristics of the gas chromatograph and the
mass detector apparatus and the column.
Table 3 — GC conditions
Column DB5 HT (15 m – 0,25 mm – 0,10 µm film thickness or 30 m – 0,25 mm – 0,25 µm film
thickness)
Injector Liner, tapered, deactivated liner with glass wool
Injection port temperature 350 °C
Injection temperature Splitless in pressure pulse (12 psi) for 1 min
program
1 min after the injection, split at 150 ml/min
3 min after the injection, split at 20 ml/min
Injection volume 1,0 µl
Oven temperature 50 °C for 1 min, ramp at 15 °C/min to 100 °C, ramp at 40 °C/min to 290 °C, ramp at 60 °C/
min to 350 °C, hold for 2 min
Carrier gas Helium/hydrogen constant flow rate, 1,2 ml/min
Syringe solvent clean-up Acetonitrile
Table 4 — MS conditions
Ion source temperature 300 °C
Quadrupole temperature 150 °C
Transfer line temperature 325 °C
Phthalates Quantifier ion Qualifier ion Qualifier ion Qualifier ion
Parameters for SIM mode DEHP 149 167 279 132
DEHP d4 153 171 283 136
Check that the volume of acetonitrile when injected at 350 °C is not greater than the volume of the liner.

4.7 Quantitative determination
4.7.1 Calibration curve
The calibration curve of DEHP is of type y = a * x
A C
ph cal-ph
=a*
A C
dph cal-dph
The calibration curve is determined as follows:
AC *
ph cal-dph
a=
AC *
dphcal-ph
where
a is the slope;
A is the peak area of DEHP in the calibration solution;
ph
A is the peak area of the DEHP-d4 internal standard in the calibration solution;
dph
C
is the concentration of the DEHP-d4 internal standard in the calibration solution, in micrograms
cal-dph
per millilitres (see Table 2);
C
is the concentration of DEHP in the calibration solution, in micrograms per millilitres (see Table 2).
cal-ph
4.7.2 Quantitative determination of DEHP
AV ** C
 
ph dphdph
C = - C
 
ph blank
 
AM ** a
dph
 
where
a is the slope;
A is the peak area of DEHP;
ph
A is the peak area of the DEHP-d4 internal standard;
dph
C is the concentration of the DEHP-d4 internal standard solution, in micrograms per millilitres;
dph
V is the volume of the DEHP-d4 internal standard solution, in millilitres;
dph
C is the concentration of DEHP, in milligrams per kilogram;
ph
C is the concentration of DEHP in the blank, in milligrams per kilogram (see 4.6.1);
blank
M is the mass of the test portion, in grams.
If the volumes of the internal standard solution indicated in this method are respected, and if the same
solution of DEHP-d4 internal standard is used both for the calibration solutions and for the sample
preparation, the following simplified calculation formulae can be used for the determination of the
calibration curve and the quantification of phthalate:

4.7.2.1 Calibration curve
The calibration curve for DEHP is of type y = a * x
A
ph 40
=am**
cal-ph
A 80
dph
The calibration curve is determined as follows:
A *2
ph
a=
Am *
dphcal-ph
where
a is the slope from the calibration, using the same DEHP-d4 internal standard solution for the
samples and for the calibration;
A is the peak area of DEHP in the calibration solution;
ph
A is the peak area of the DEHP-d4 internal standard in the calibration solution;
dph
m
is the mass of DEHP in the solution calibration, in micrograms.
cal-ph
NOTE The coefficient of 2 is added in the formula to consider the ratio of the amount of internal standard between
the calibration solutions and the samples (80 µl of the internal standard solution for the calibration and 40 µl of the
internal standard solution for the samples).
4.7.2.2 Quantitative determination of DEHP
A
 
ph
C = - C
 
ph blank
 
AM ** a
 dph 
where
C is the concentration of DEHP, in milligrams per kilogram;
ph
a is the slope from the calibration, using the same DEHP-d4 internal standard solution for the
samples and for the calibration;
A is the peak area of the DEHP;
ph
A is the peak area of the DEHP-d4 internal standard;
dph
M is the mass of the test portion, in grams;
C is the concentration of DEHP in the blank, in milligrams per kilogram (see 4.6.1).
blank
4.8 Precision of the method
4.8.1 Interlaboratory test
Details of the test and the precision of the method are summarized in Annex A. The values derived from this
interlaboratory test will possibly not be applicable to concentration ranges and matrices other than those
given.
4.8.2 Repeatability
The absolute difference between two independent single test results, obtained with the same method on
identical test material in the same laboratory by the same operator using the same equipment within a short
interval of time, shall in not more than 5 % of cases exceed the value of r given in Table A.1.
4.8.3 Reproducibility
The absolute difference between two single test results, obtained with the same method on identical test
material in different laboratories by different operators using different equipment, shall in not more than
5 % of cases exceed the value of R given in Table A.1.
4.9 Test report
The test report shall contain at least the following information:
a) all information necessary for the complete identification of the sample;
b) the sampling method used, if known;
c) the test method used, with reference to this document, i.e. ISO/TS 16465:2024;
d) all operating details not specified in this document, or regarded as optional, together with details of any
incidents occurred when performing the method, which can have influenced the test result(s);
e) the test result obtained;
f) if the repeatability has been checked, the final quoted result obtained.
5 Part B - Determination of eight phthalates
5.1 Principle
Phthalates are first extracted from oil with acetonitrile followed by dispersive-solid-phase extraction
clean-up (dispersive SPE) which is done to remove organic acids, excess water and other components
using a combination of primary secondary amine (PSA) sorbent and C18 bonded phase. Finally, the extract
is concentrated and analysed by gas chromatography-mass spectrometry (GC-MS). For refined oil, the
acetonitrile extract is directly concentrated without using the dispersive-SPE purification step.
5.2 Reagents
WARNING — Attention is drawn to national regulations that specify the handling of hazardous
substances, and users' obligations thereunder. Technical, organizational and personal safety
measures shall be followed.
Unless otherwise specified, use only reagents of recognized analytical grade.
5.2.1 Toluene, purity 99 % min.
5.2.2 Acetonitrile, trace organic analysis grade, purity 99 % min.
5.2.3 n-Hexane, trace organic analysis grade, purity 99 % min.
3)
5.2.4 Chromabond adsorbent Diamino (PSA), Chromabond adsorbent C18 .
3) Chromabond sorbent C18 ref 730611 and Chromabond sorbent Diamino ref 730611 from Macherey Nagel
(www .mn -net .com) have proved to be free of phthalate contamination. This information is given for the convenience of
users of this document and does not constitute an endorsement by ISO of these products.
...

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ISO/TS 16465:2024 표준은 식물성 기름에서 프탈레이트의 정량적인 측정을 위한 두 가지 방법을 규정하고 있습니다. 이 표준의 범위는 다이-2-에틸헥실 프탈레이트(DEHP) 측정을 위한 방법 A와, 다양한 프탈레이트(디메틸 프탈레이트(DMP), 다이에틸 프탈레이트(DEP), 다이-이소부틸 프탈레이트(DIBP), 다부틸 프탈레이트(DBP), 벤질부틸 프탈레이트(BBP), 다이-2-에틸헥실 프탈레이트(DEHP), 다이-이소노닐 프탈레이트(DINP), 다이-이소데실 프탈레이트(DIDP))를 측정하기 위한 방법 B로 나뉘어 있습니다. 이 두 가지 방법은 원유, 정제유, 버진 오일을 포함한 모든 식물성 기름에 적용 가능하여 그 응용 범위가 넓습니다. ISO/TS 16465:2024의 강점은 GC-MS(가스크로마토그래피-질량분석기)를 이용한 고도한 분석 방법을 통해 프탈레이트의 정확한 농도를 정량적으로 측정할 수 있다는 것입니다. 이 표준은 식물성 기름의 품질 관리와 안전성을 담보할 수 있는 중요한 도구로, 식품 산업에 필수적인 지침을 제공합니다. 특히, 다양한 식물성 기름에 대한 적합성을 고려하여 설정된 여러 프탈레이트의 측정 방법은 사용자에게 실질적인 이점을 제공합니다. 이러한 이유로 ISO/TS 16465:2024는 식품 안전 및 품질 관리를 중요시하는 산업 관계자들에게 매우 관련성이 높고 유용한 표준입니다. 프탈레이트와 같은 오염 물질의 관리를 통해 소비자에게 안전한 제품을 제공하는 데 기여할 것으로 기대됩니다.

Le document ISO/TS 16465:2024 traite de la détermination des phtalates dans les huiles végétales, spécifiant deux méthodes distinctes de quantification par chromatographie en phase gazeuse-spectrométrie de masse (GC-MS). Cette norme est d'une grande pertinence, notamment pour les professionnels du secteur agroalimentaire et de l'analyse des huiles, car elle répond à un besoin croissant de contrôler les contaminants chimiques dans les produits alimentaires. La première méthode, Part A, se concentre sur la détermination du phtalate de di-2-éthylhexyle (DEHP), un composé préoccupant en raison de ses effets potentiels sur la santé humaine. La seconde méthode, Part B, élargit l'analyse à un éventail de huit phtalates, dont le diméthyl phtalate (DMP) et le dibutyl phtalate (DBP). La capacité des deux méthodes à être appliquées à tous types d'huiles végétales, qu'elles soient brutes, raffinées ou vierges, renforce leur utilité dans un contexte commercial où la diversité des produits est la norme. Parmi les points forts de la norme ISO/TS 16465:2024, on trouve la clarté des protocoles décrits, qui facilitent leur mise en œuvre par des laboratoires et des industriels. La précision apportée par la méthode GC-MS permet d'obtenir des résultats fiables et reproductibles, ce qui est essentiel pour satisfaire aux exigences réglementaires et assurer la sécurité des consommateurs. De plus, le fait que cette norme soit spécifiquement axée sur les huiles végétales souligne son importance dans un marché où la consommation d'huiles issues de sources végétales ne cesse d'augmenter. En somme, l'ISO/TS 16465:2024 est une norme incontournable pour la détermination des phtalates dans les huiles végétales, garantissant des méthodes adaptées et robustes pour une analyse précise, essentielle pour la conformité réglementaire et la protection des consommateurs.

ISO/TS 16465:2024は、植物油におけるフタル酸エステルの定量的な決定に関する重要な標準です。この文書は、ガスクロマトグラフィー-質量分析法(GC-MS)を使用した二つの方法を具体的に定義しています。第一部では、ジ2-エチルヘキシルフタレート(DEHP)の決定方法が示されており、第二部では、ジメチルフタレート(DMP)、ジエチルフタレート(DEP)、ジイソブチルフタレート(DIBP)、ジブチルフタレート(DBP)、ベンジルブチルフタレート(BBP)、ジ2-エチルヘキシルフタレート(DEHP)、ジイソノニルフタレート(DINP)、ジイソデシルフタレート(DIDP)の8種類のフタル酸エステルを対象としています。 この標準の強みは、すべての植物油-未精製、精製、バージンを含む-に適用可能であることです。また、フタル酸エステルに関連する安全性の問題が増加している中で、ISO/TS 16465:2024は、業界のニーズに応える形で科学的な根拠に基づいた方法を提供します。特に、消費者の健康と環境保護が重要視されている現代において、この標準の関連性は高まっています。 標準は、分析精度や再現性を確保するための明確な指針を提供し、研究者や業界専門家が信頼性のあるデータを得るための手助けになります。これにより、植物油の品質管理および製品の安全性保証に寄与することが期待されます。ISO/TS 16465:2024は、フタル酸エステルに関する規制遵守を推進し、植物油市場における透明性を高めるための重要なツールです。

Die Norm ISO/TS 16465:2024 beschäftigt sich mit der Bestimmung von Phthalaten in pflanzlichen Ölen und stellt zwei präzise Methoden für die quantitative Analyse dieser Stoffe durch Gaschromatographie-Massenspektrometrie (GC-MS) bereit. Der Umfang dieser Norm ist umfassend, da sie sowohl die Bestimmung von Di-2-ethylhexyl phthalat (DEHP) in Teil A als auch die gleichzeitige Analyse von acht weiteren Phthalaten in Teil B umfasst. Diese Phthalate umfassen dimethyl phthalate (DMP), diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-isononyl phthalate (DINP) und di-isodecyl phthalate (DIDP). Ein wesentlicher Vorteil dieser Norm ist die breite Anwendbarkeit der Methoden auf alle Arten von pflanzlichen Ölen, einschließlich Roh-, Raffinade- und Nativölen. Dies macht ISO/TS 16465:2024 zu einem wichtigen Instrument für Hersteller und Laboratorien, die sicherstellen möchten, dass ihre Produkte den aktuellen Sicherheits- und Gesundheitsstandards entsprechen. Die Relevanz der Norm ergibt sich aus der zunehmenden Regulierung und Überwachung von Phthalaten in Lebensmitteln und kosmetischen Produkten. Angesichts der Bedenken hinsichtlich der Auswirkungen dieser Chemikalien auf die Umwelt und die menschliche Gesundheit ist die Fähigkeit, Phthalate in pflanzlichen Ölen präzise zu bestimmen, von entscheidender Bedeutung. Die Standardisierung stellt sicher, dass die Ergebnisse zwischen verschiedenen Laboren reproduzierbar sind, was die Qualität und Sicherheit von pflanzlichen Ölen erheblich erhöht. Insgesamt bietet die Norm ISO/TS 16465:2024 eine fundierte Grundlage für die Analyse von Phthalaten in pflanzlichen Ölen und stärkt somit die Einhaltung von Sicherheitsstandards in der Lebensmittelindustrie.

ISO/TS 16465:2024 provides a comprehensive framework for the quantitative determination of phthalates in vegetable oils, addressing a crucial aspect of food safety and quality in the oils industry. The standard delineates two specific methodologies using gas chromatography-mass spectrometry (GC-MS) which enhance the reliability and accuracy of phthalate detection in various types of vegetable oils, including crude, refined, and virgin oils. Strengths of this standard are particularly evident in its dual approach. Part A focuses specifically on the determination of di-2-ethylhexyl phthalate (DEHP), a prevalent phthalate often found in materials that come into contact with food products. This targeted method ensures that one of the most common contaminants is effectively monitored. Part B expands the analysis to include seven additional phthalates (dimethyl phthalate (DMP), diethyl phthalate (DEP), di-isobutyl phthalate (DIBP), dibutyl phthalate (DBP), benzylbutyl phthalate (BBP), di-isononyl phthalate (DINP), and di-isodecyl phthalate (DIDP)), thereby providing a holistic approach to assessing phthalate contamination in vegetable oils. By doing so, the standard facilitates compliance with safety regulations and industry best practices, ultimately contributing to consumer protection. The relevance of ISO/TS 16465:2024 is underscored by the increasing scrutiny surrounding food safety and the potential health impacts of phthalate exposure. As more consumers become aware of the presence of chemical contaminants in their food sources, the standard serves as a vital tool for manufacturers and regulators who strive to uphold high safety and quality standards within the food supply. In summary, ISO/TS 16465:2024 stands out as a significant advancement in the field of food safety, offering clear methodologies that are both applicable and essential for the determination of phthalates in vegetable oils. The standard's comprehensive scope ensures that all relevant phthalates are addressed, making it an indispensable resource for industry stakeholders committed to ensuring the safety and quality of vegetable oil products.