Surface active agents — Detergents — Determination of alkylphenol ethoxylates
This document provides a method for the determination of alkylphenol ethoxylates (APEOs) in surfactants using high performance liquid chromatography (HPLC) and detected with diode array detector (DAD) or fluorescence detector (FLD). This method is appropriate for the detection and quantification of APEOs in surfactants.
Agents de surface — Détergents — Dosage des alkylphénols éthoxylés
Standards Content (Sample)
Surface active agents — Detergents
— Determination of alkylphenol
Agents de surface — Détergents — Dosage des alkylphénols éthoxylés
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1 Scope . 1
2 Normative references . 1
3 Terms and definitions . 1
4 Principle . 1
5 Reagents and materials . 1
6 Apparatus . 2
7 Procedure. 2
7.1 Standard preparation . 2
7.2 Sample preparation . 2
7.3 Analysis. 3
7.3.1 Chromatographic condition . 3
7.3.2 Qualitative determination . 3
7.3.3 Calibration curve . 3
7.3.4 Quantitative determination . 4
8 Expression of results . 4
8.1 Calculation . 4
8.2 E valuation . 4
8.3 Precision . 5
8.3.1 Repeatability . 5
8.3.2 Reproducibility limit. 5
9 Test report . 5
Annex A (informative) Chromatogram of alkylphenol ethoxylates (APEOs) .6
Annex B (informative) Identification test . 7
Annex C (informative) Statistical data from the ring carried out . 9
© ISO 2019 – All rights reserved iii
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INTERNATIONAL STANDARD ISO 21264:2019(E)
Surface active agents — Detergents — Determination of
This document provides a method for the determination of alkylphenol ethoxylates (APEOs) in
surfactants using high performance liquid chromatography (HPLC) and detected with diode array
detector (DAD) or fluorescence detector (FLD).
This method is appropriate for the detection and quantification of APEOs in surfactants.
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 607, Surface active agents and detergents — Methods of sample division
3 Terms and definitions
No terms and definitions are listed in this document.
ISO and IEC maintain terminological databases for use in standardization at the following addresses:
— ISO Online browsing platform: available at https: //www .iso .org/obp
— IEC Electropedia: available at http: //www .electropedia .org/
The sample is weighed into a vial and extracted in methanol using an ultrasonic bath. Subsequently
the extract is filtered and analysed using high performance chromatography system with diode array
detector (DAD) or fluorescence detector (FLD). Quantification is achieved by external quantification
method and using the relationship chromatographic peak through its unique its retention time.
5 Reagents and materials
During the analysis, use only reagents of recognized analytical grade and the water of quality for HPLC
5.1 Methanol, HPLC Grade.
5.2 Octylphenol ethoxylates (OPEO, 2 ≤ n ≤ 12), for example, Triton X-100 , CAS no. 9002-93-1,
Sigma-Aldrich Part Number T9284.
1) Triton X-1000 ®, CAS no. 9002-93-1, Sigma Aldrich Part Number T9284 is an example of products. 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.
© ISO 2019 – All rights reserved 1
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5.3 Nonylphenol ethoxylates (NPEO, 3 ≤ n ≤ 18), for example, IGEPAL CO-630 , CAS no. 9016-45-
9, Sigma-Aldrich Part Number 542334.
NOTE There is a possibility that reagents of the grade of OPEO and NPEO are not available.
Normal laboratory apparatus and, in particular, the following:
6.1 Analytical balance, weighing to an accuracy of 0,1 mg.
6.2 Ultrasonic bath, 40 kHz, with thermostat.
6.3 Membrane filter, polyamide, 0,45 μm.
6.4 Volumetric flasks, 10 ml, 50 and 100 ml capacity.
6.5 High-performance Liquid Chromatography (HPLC) with DAD or FLD detector.
6.6 Reverse phase column with guard column, a stainless-steel column 150 mm and 2,1 mm in
internal diameter filled with 3,5 µm ODS (C ) as stationary phases.
7.1 Standard preparation
7.1.1 Prepare 1 000 mg/l stock solutions of each APEOs from standard materials or certified solutions in
methanol. 100 mg of the OP EO (5.2) and NP EO (5.3) are dissolved in different 100 ml volumetric flasks
with methanol and filled up to the mark respectively.
Depending on the stock concentrations prepared, the solubility at that concentration will have to be
7.1.2 Prepare calibration curve for samples of each APEO stock standard solution at required
concentrations by adding aliquots of stock solutions to a 100 ml volumetric flask and then diluted with
methanol to the desired calibration level. If lower reporting limit is required, lower concentration of
calibration standard shall be prepared to cover the reporting limit. Each calibration curve is prepared
using the respective stock solution. The preparation of the Level 5 standard can be accomplished using
different volumes and concentrations of stock solutions as is accustomed in the individual laboratory.
The calibration vials shall be used within 24 h to ensure optimum results. Stock calibration standards
are routinely repla