GAFTI Analytical Method

Detection and Determination of

Alkylphenols (APs) and Alkylphenol
Ethoxylates (APEOs) in Textiles, Leathers
and Plastics by LC-MS or LC-MS-MS

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GAFTI Analytical Method
Detection and Determination of
Alkylphenols (APs) and Alkylphenol
Ethoxylates (APEOs) in Textiles, Leathers
and Plastics by LC-MS or LC-MS-MS

1. Scope

This method is for the detection and quantification of alkylphenols (APs) and
alkylphenol ethoxylates (APEOs) in textiles, leathers and plastics.

2. Principle

Extraction of APEOs and APs in textile and leather is carried out with
methanol. The extract is analyzed by HPLC-MS. For plastic material,
tetrahydrofuran is used for the extraction and then precipitated with
acetonitrile. The extract is then analyzed by HPLC-MS. Either MRM or
qualifier ion determination could be used for confirmation of APs.

3. Reagents

3.1. Nonylphenol ethoxylates (NPEOs) (IGEPAL CO-630), CAS no. 68412-

54-4, Sigma-Aldrich Part Number 542334.

3.2. Octylphenol ethoxylates (OPEOs) (Triton X-100), CAS no. 9002-93-1,

Sigma-Aldrich Part Number T9284.

3.3. Nonylphenols (mixture) (NPs), CAS no. 84852-15-3, Sigma-Aldrich Part

number 290858.

3.4. 4-n-Nonylphenol (4-n-NP), CAS no. 104-40-5, Sigma-Aldrich Part

number 46405.

3.5. 4-n-Octylphenol (4-n-OP), CAS no. 1806-26-4, Aldrich Part number

384445.

3.6. 4-tert-Octylphenol (4-tert-OP), CAS no. 140-66-9, Aldrich Part number

290823.

3.7. Methanol, HPLC grade.

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3.8. Tetrahydrofuran (THF), HPLC grade (inhibitor free). Note: BHT as
inhibitor in solvent will interfere the peak of APs.

3.9. Acetonitrile (ACN), HPLC grade.

3.10. MilliQ water.

3.11. Ammonium acetate.

3.12. 0.5 M Ammonium acetate.

Dissolve 9.635 g of ammonium acetate in 250 ml of MilliQ water.

3.13. 5 mM Ammonium acetate.

Pipette 5 ml of 0.5 M ammonium acetate solution (Clause 3.12) into a
500 ml volumetric flask and fill up to the mark with MilliQ water.

3.14. 5 mM Ammonium acetate in ACN.

Pipette 5 ml of 0.5 M ammonium acetate solution (Clause 3.12) into a
500 ml volumetric flask and fill up to the mark with ACN.

4. Apparatus

4.1. General laboratory apparatus.

4.2. Liquid Chromatograph Mass Spectrometer (LC-MS).

4.3. Ultrasonic bath to be set up at 70 oC 5 oC.

4.4. 0.45 m nylon filter disc.

5. Sample preparation

5.1. Standard preparation.

5.1.1. Prepare 1000 mg/L stock solutions of each APEOs and APs analyte
in methanol.

5.1.2. Prepare calibration curve for textile and leather materials with
calibrators of each APEOs and APs at 0.1, 0.5, 1, 5 mg/L* in
methanol.

5.1.3. Prepare calibration curve for plastic material with calibrators of each
APEOs and APs at 0.1, 0.5, 1, 5 mg/L* in 2:1 ACN:THF.

*If lower reporting limit is required, lower concentration of calibration

standard shall be prepared to cover the reporting limit.

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5.2. Sample preparation for textile (including textile with printing) and leather.

5.2.1. Cut the sample into small pieces and mix homogeneously, textile 5
mm x 5 mm; leather 4 mm x 4 mm.

5.2.2. Weigh 1 g of sample into an extraction vial.

5.2.3. Pipette 20 ml of methanol into the extraction vial and ensure all
sample is soaked in the solution.

5.2.4. Place the vial into an ultrasonic bath at 70 oC for 1 hour.

5.2.5. Cool the extract to room temperature and then filter through a 0.45 m
nylon filter disc into a HPLC vial for analysis.

5.3. Sample preparation for plastic (including synthetic leather).

5.3.1. Cut the sample into small pieces (2 mm x 2 mm) and mix
homogeneously.

5.3.2. Weigh 1 g of sample into an extraction vial.

5.3.3. Pipette 10 ml of THF into the extraction vial and ensure all sample is
soaked in the solution.

5.3.4. Place the vial into an ultrasonic bath at 70 oC for 1 hour.

5.3.5. Pipette 20 ml of ACN into the extraction vial and shake vigorously for
precipitation.

5.3.6. Stay for 10 minutes and then filter the solution through a 0.45 m
nylon filter disc into a HPLC vial for analysis.

6. LC-MS conditions

6.1. Chromatographic conditions

Zorbax Eclipse XDB-C18, 2.1 mm x 150 mm, 3.5 m

Column
or equivalent
Injection volume 10 l
Flow 0.3 ml/min
Column
40 oC
temperature
Eluent A 5 mM ammonium acetate (Clause 3.13)
Eluent B 5 mM ammonium acetate in ACN (Clause 3.14)

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 Gradient program:

Time (min) Eluent A (%) Eluent B (%)

0 50 50
10 2 98
15 2 98
15.5 50 50
20 50 50

6.2. SIM mass of APEOs (Positive Polarity)

NPEO Target mass (m/z) OPEO Target mass (m/z)

congeners [M + NH4] congeners [M + NH4]
NPEO 18 1030 OPEO 16 928
NPEO 17 986 OPEO 15 884
NPEO 16 942 OPEO 14 840
NPEO 15 898 OPEO 13 796
NPEO 14 854 OPEO 12 752
NPEO 13 810 OPEO 11 708
NPEO 12 766 OPEO 10 664
NPEO 11 722 OPEO 9 620
NPEO 10 678 OPEO 8 576
NPEO 9 634 OPEO 7 532
NPEO 8 590 OPEO 6 488
NPEO 7 546 OPEO 5 444
NPEO 6 502 OPEO 4 400
NPEO 5 458 OPEO 3 356
NPEO 4 414 OPEO 2 312
NPEO 3 370
NPEO 2 326

6.3. SIM mass of APs (Negative Polarity) For qualification and

quantification

Compounds Target mass (m/z)

4-n-NP 219
NP (mixture) 219
4-n-OP 205
4-tert-OP 205

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6.4. Confirmation of APs

Either one of following techniques could be adopted to confirm the

identity of APs detected in sample solution.

6.4.1. MRMs of APs (Negative Polarity)

Compounds M1 (m/z) M2 (m/z) M3 (m/z)

4-n-NP 219 106 --
NP (mixture) 219 133 117
4-n-OP 205 106 --
4-tert-OP 205 133 117

M1 is the molecular weight of the parent ion

M2 is the molecular weight of the most abundant daughter ion
M3 is the molecular weight of the second most abundant daughter ion

6.4.2. Qualifier ion of APs (Negative Polarity)

First qualifier Second qualifier

Compounds
ion (m/z) ion (m/z)
4-n-NP 106 --
NP (mixture) 133 117
4-n-OP 106 --
4-tert-OP 133 117

7. Calculation of results

7.1. Calculation of APEOs result.

7.1.1. Determination of the R value for each congeners of APEO.

Calibrate of weight fraction (R) for each APEO congeners and

calculate concentration in each APEO congeners as follows:

For example,

Where,

AO = Area response of each OPEO congeners

AOT = Total sum of OPEOs area response (from OPEO 2
OPEO 16)

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 Exact concentration (C) in
each OPEO (n) (mg/l)

Where,

Cstd = Concentration of OPEO standard mix solution in

each working solution (Clause 5.1.2 or 5.1.3)

7.1.2. Calibration curve

Prepare calibration curves by plotting the concentration of each APEO

congeners against R value.

For quantification, the calibration curve shall have a correlation

coefficient better than or equal to 0.995. (i.e. R2 = 0.99)

7.1.3. Calculation of concentration of APEOs result.

The APEO level is calculated in sample according to the following

equation:

Where,

[C] = Total sum of concentration of each NPEO/OPEO

congeners (mg/l)
V = Extraction volume (ml)
W = Weight of the sample (g)
D.F. = Dilution factor

7.2. Calculation of APs result.

Where,

C = Concentration of APs in extract (mg/l)

V = Extraction volume (ml)
W = Weight of the sample (g)
D.F. = Dilution factor

7.3. Reporting limit

Reporting limit of this method is 10 mg/kg.

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8. Test report

The test report shall contain at least the following information:

a) all information necessary for the identification of the sample

b) data and type of sampling (if known)

c) date of receipt of the laboratory sample

d) date of test

e) the results and the units in which the results have been expressed

f) any particular points observed in the course of the test

g) any operations not specified in the method or regarded as optional which

might have affected the results

9. Reference

ISO/DIS 18254 Textiles Method for the detection and determination of

alkylphenolethoxylates (APEO)

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