Supporting Information
Analytical characterization and comparison of
tristyrylphenol ethoxylates used in agrochemical
formulation
Johannes Glaubitz1,2, Torsten C. Schmidt1,*
1 University Duisburg-Essen, Instrumental Analytical Chemistry, Universitätsstr. 5,
45141 Essen, Germany
2 Bayer CropScience, Formulation Technology Analysis & Services, Alfred-Nobel-Straße 50,
40789 Monheim am Rhein, Germany
*Corresponding author:
Phone: +49 201 183-6774
Fax: +49 201 183-6773
E-mail: torsten.schmidt@uni-due.de
Table of Contents
1.
Sample for testing on mass calibration of TOF-MS .................................................................................. 5
2.
Mass spectra of the major compounds in TSP-16 .................................................................................... 7
3.
Comparison of the ionization performance of APPI and ESI for the analysis of TSP-40-ethoxylates ...... 10
4.
Determination of the limit of quantification ......................................................................................... 12
5.
Data bases and results files for the data mining .................................................................................... 16
6.
Results of the principal components analysis according to the score of the single compounds ............. 16
7.
Comparison of TSP-16 of different suppliers and qualities .................................................................... 19
8.
Statistical evaluation of the results on the content of the components in TSP-16 on their use for product
identification .................................................................................................................................................. 22
9.
Example for interference on analysis of TSP-16 in agrochemical formulations ...................................... 24
1
List of Figures
Figure S 1: The mass spectra of the major compounds are displayed in Figure S 1 (a) for PEG,
in Figure S 1 (b) for MSP ethoxylates, in Figure S 1 (c) for DSP ethoxylates, in Figure S 1 (d)
for TSP ethoxylates and in Figure S 1 (e) for TeSP ethoxylates. These mass spectra were
obtained using APCI as coupling to the TOF mass spectrometer .............................................. 7
Figure S 2: Ionization behavior of TSP-40 ionized by APPI (a) and ESI (b). In each case the
mass spectrum over the peak of TSP-ethoxylates is displayed. For each experiment the same
elution conditions with water and methanol as mobile phase, plus 5 mmol/L ammonium
formiate, were chosen. The mass spectrometer used for this experiments was a Thermo Qexactive. .................................................................................................................................... 11
Figure S 3: Chromatograms for determination of the signal-to-noise ratio at the defined LOQ
level for TSP with 16 EO units (a) and hexanophenone (b). The LOQ was defined as a signalto-noise ratio of at least 20:1, which has been achieved for both analytes. ............................. 13
Figure S 4: Linear ranges for TSP with 16 EO units (a) and hexanophenone (b) including the
bands of prediction indicated green for the upper and red for the lower limit ......................... 14
Figure S 5: Distribution of residues for the regression analysis of standards of TSP with 16
EO units a) and the internal standard hexanophenone b) ......................................................... 15
Figure S 6: Principle component analysis of the data sets from supplier A (Cross), B1
(Arrow), B2 (Horizontal Bar) and C (Vertical bar) together with the data of the formulation
samples containing TSP-16 of supplier A (Square), B1 (Diamond), B2 (Circle) and C
(Triangle). For the PCA the whole data set was taken including the 3 repetition analysis each
production batch and formulation sample. ............................................................................... 22
Figure S 7: Combined hierarchical clustering of the samples (x-axis) and the compounds (yaxis) detected in the samples of supplier A, B1, B2 and C together with sample of formulation
2
containing TSP-16 of Supplier A, B1, B2 and C. Each sample is the average of 3 repetition
analyses. The content of a compound in the analyzed sample is coded via black-white
rectangles in the column beneath the respective samples. The color ranges from black,
compound not detected, over grey, compound as abundant as internal standard, to white,
compound with the maximum content. .................................................................................... 23
Figure S 8: Extracted ion chromatograms obtained in the positive ionization mode of terminal
phosphated (a) and sulfated (b) commercially available TSP-16. Indicated are the identified
entities of DSP-, TSP and TeSP ethoxylates. ........................................................................... 25
Figure S 9: Principal component analysis of the data sets from supplier A (Cross), B1
(Arrow), B2 (Horizontal Bar) and C (Vertical bar) together with the data of the formulation
samples containing TSP-16 of supplier A (Square), B2 (Circle) and C (Triangle). For the PCA
the whole data set was taken including the 3 repetition analysis each production batch and
formulation sample. .................................................................................................................. 26
3
List of Tables
Table S 1: Retention time and exact masses for compounds in the test sample for checking on
mass calibration .......................................................................................................................... 5
Table S 2: Results for the test on heteroscedasticity against a level of significance of 0.05 on
the data set used for linear regression of TSP-with 16 EO units and the internal standard
hexanophenone according to Breusch-Pagan ........................................................................... 15
Table S 3: Given are the scores each compound for component 1 and 2 of the principle
component analysis performed on the data set of the different suppliers of TSP-16, as it is
graphically displayed in Figure 6 in the manuscript. ............................................................... 16
Table S 4: Compounds used for the combined hierarchical clustering listed together with the
corresponding arrays as defined in Figure 7 (see manuscript). The compounds are sorted
according to the order obtained by the hierarchical clustering of the compounds. .................. 19
Table S 5: Table of composition of the model agrochemical formulation containing terminal
sulfated TSP-16 alongside with TSP-16 .................................................................................. 25
4
1. Sample for testing on mass calibration of TOF-MS
The retention times and exact masses for the compounds in the test sample for checking mass
calibration of the used TOF-MS are given in Table S 1.
Table S 1: Retention time and exact masses for compounds in the test sample for checking on mass
calibration
Compound
Imidacloprid
Thiacloprid
Tebuconazole (1.Isomer)
Triadimenol
Tebuconazole (2.Isomer)
Distyrylethoxylate-5-EO
Distyrylethoxylate-6-EO
Distyrylethoxylate-7-EO
Distyrylethoxylate-8-EO
Distyrylethoxylate-9-EO
Distyrylethoxylate-10-EO
Distyrylethoxylate-11-EO
Distyrylethoxylate-12-EO
Distyrylethoxylate-13-EO
Distyrylethoxylate-14-EO
Distyrylethoxylate-15-EO
Distyrylethoxylate-16-EO
Distyrylethoxylate-17-EO
Distyrylethoxylate-18-EO
Distyrylethoxylate-19-EO
Distyrylethoxylate-20-EO
Distyrylethoxylate-21-EO
Distyrylethoxylate-22-EO
Distyrylethoxylate-23-EO
Distyrylethoxylate-24-EO
Distyrylethoxylate-25-EO
Distyrylethoxylate-26-EO
Distyrylethoxylate-27-EO
Distyrylethoxylate-28-EO
Distyrylethoxylate-29-EO
Distyrylethoxylate-30-EO
Nonylphenolethoxylate-5-EO
Nonylphenolethoxylate-6-EO
Nonylphenolethoxylate-7-EO
Nonylphenolethoxylate-8-EO
Nonylphenolethoxylate-9-EO
Nonylphenolethoxylate-10-EO
Nonylphenolethoxylate-11-EO
Nonylphenolethoxylate-12-EO
Nonylphenolethoxylate-13-EO
Nonylphenolethoxylate-14-EO
Nonylphenolethoxylate-15-EO
tN [min]
2.0
2.5
4.3
4.6
4.9
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.8
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
6.6
6.3
6.2
6.2
6.2
6.2
6.2
6.2
6.2
6.2
6.2
Exact mass [m/z]
254.0450
252.0236
307.1451
295.1088
307.1451
522.2981
566.3244
610.3506
654.3768
698.4030
742.4292
786.4554
830.4816
874.5079
918.5341
962.5603
1006.5865
1050.6127
1094.6389
1138.6651
1182.6914
1226.7176
1270.7438
1314.7700
1358.7962
1402.8224
1446.8486
1490.8749
1534.9011
1578.9273
1622.9535
440.3138
484.3400
528.3662
572.3924
616.4186
660.4449
704.4711
748.4973
792.5235
836.5497
880.5759
5
Compound
Nonylphenolethoxylate-16-EO
Nonylphenolethoxylate-17-EO
Nonylphenolethoxylate-18-EO
Nonylphenolethoxylate-19-EO
Nonylphenolethoxylate-20-EO
Nonylphenolethoxylate-21-EO
Nonylphenolethoxylate-22-EO
Nonylphenolethoxylate-23-EO
Nonylphenolethoxylate-24-EO
Nonylphenolethoxylate-25-EO
Nonylphenolethoxylate-26-EO
Nonylphenolethoxylate-27-EO
Nonylphenolethoxylate-28-EO
Nonylphenolethoxylate-29-EO
Nonylphenolethoxylate-30-EO
Tristyrylethoxylate-5-EO
Tristyrylethoxylate-6-EO
Tristyrylethoxylate-7-EO
Tristyrylethoxylate-8-EO
Tristyrylethoxylate-9-EO
Tristyrylethoxylate-10-EO
Tristyrylethoxylate-11-EO
Tristyrylethoxylate-12-EO
Tristyrylethoxylate-13-EO
Tristyrylethoxylate-14-EO
Tristyrylethoxylate-15-EO
Tristyrylethoxylate-16-EO
Tristyrylethoxylate-17-EO
Tristyrylethoxylate-18-EO
Tristyrylethoxylate-19-EO
Tristyrylethoxylate-20-EO
Tristyrylethoxylate-21-EO
Tristyrylethoxylate-22-EO
Tristyrylethoxylate-23-EO
Tristyrylethoxylate-24-EO
Tristyrylethoxylate-25-EO
Tristyrylethoxylate-26-EO
Tristyrylethoxylate-27-EO
Tristyrylethoxylate-28-EO
Tristyrylethoxylate-29-EO
Tristyrylethoxylate-30-EO
tN [min]
6.2
6.2
6.2
6.2
6.2
6.2
6.2
6.2
6.2
6.2
5.9
5.9
5.9
5.9
5.9
5.9
5.9
5.9
6.5
5.9
5.9
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
6.0
5.9
5.9
5.9
5.9
5.9
5.8
5.8
5.8
5.8
5.8
Exact mass [m/z]
924.6022
968.6284
1012.6546
1056.6808
1100.7070
1144.7332
1188.7594
1232.7857
1276.8119
1320.8381
1364.8643
1408.8905
1452.9167
1496.9429
1540.9692
626.3607
670.38695
714.4132
758.4394
802.4656
846.4918
890.5180
934.5442
978.5705
1022.5967
1066.6229
1110.6491
1154.6753
1198.7015
1242.7278
1286.7540
1330.7802
1374.8064
1418.8326
1462.8588
1506.8850
1550.9113
1594.9375
1638.9637
1682.9899
1727.0161
6
2. Mass spectra of the major compounds in TSP-16
In the following Figure S 1 the mass spectra of the major compounds in TSP-16 are shown. In
Figure S 1 (a) the mass spectrum of PEG, in Figure S 1 (b) the mass spectrum of MSP
ethoxylates, in Figure S 1 (c) the mass spectrum of DSP ethoxylates, in Figure S 1 (d) the
mass spectrum of TSP ethoxylates and in Figure S 1 (e) the mass spectrum of TeSP (4)
ethoxylates are given.
Figure S 1: The mass spectra of the major compounds are displayed in Figure S 1 (a) for PEG, in Figure S
1 (b) for MSP ethoxylates, in Figure S 1 (c) for DSP ethoxylates, in Figure S 1 (d) for TSP ethoxylates and
in Figure S 1 (e) for TeSP ethoxylates. These mass spectra were obtained using APCI as coupling to the
TOF mass spectrometer
Signal intensity [counts]
PEG
PEG with 9 EO-units [M + NH4]
Mass-to-Charge [m/z]
+
(a)
7
MSP with 16 EO-units [M + NH4]
+
Signal intensity [counts]
(1) MSP-ethoxylates
PEG with 4 EO-units [M + NH4]
+
Mass-to-Charge [m/z]
DSP with 15 EO-units [M + NH4]
(2) DSP-ethoxylates
+
+
Signal intensity [counts]
PEG with 4 EO-units [M + NH4]
(b)
Mass-to-Charge [m/z]
(c)
8
(3) TSP-ethoxylates
(3) TSPTSP with 15 EO-units [M + NH ]
ethoxylat
+
4
Signal intensity [counts]
PEG with 4 EO-units [M + NH4]
+
Mass-to-Charge [m/z]
TeSP with 13 EO-units [M + NH4]
(d)
+
Signal intensity [counts]
(4) TeSP-ethoxylates
PEG with 5 EO-units [M + NH4]
+
Mass-to-Charge [m/z]
(e)
As shown all compounds were identified as [M + NH4]-adducts, due to the addition 5mM of
ammonium formate to the eluents. Furthermore, were identified entities of PEG for each of
the styrenated phenol ethoxylates. This PEG is due to insource degradation of the respective
9
styrenated phenol ethoxylates, as PEG originating from the sample of TSP-16 has been
chromatographically separated and is eluting before the styrenated phenol ethoxylates
between tR=1.0 min and tR=7.0 min.
3. Comparison of the ionization performance of APPI and ESI for the analysis
of TSP-40-ethoxylates
The ionization performance of APPI and ESI was compared for the analysis of TSP-40ethoxylates. For comparison the mass spectra of TSP-ethoxylates were taken for each
ionization technique. Results for APPI are displayed in Figure S 2 (a) and for ESI in Figure S 2
(b).
+
Relative abundance [%]
TSP with 33 EO-units [M + Na]
Mass-to-Charge [m/z]
(a)
10
TSP with 39 EO-units [M + 3NH4]3+
TSP with 38 EO-units [M + 2NH4]2+
Relative abundance [%]
TSP with 43
EO-units
[M + 4NH4]4+
TSP with 35 EO-units [M + NH4]+
Mass-to-Charge [m/z]
(b)
Figure S 2: Ionization behavior of TSP-40 ionized by APPI (a) and ESI (b). In each case the mass
spectrum over the peak of TSP-ethoxylates is displayed. For each experiment the same elution conditions
with water and methanol as mobile phase, plus 5 mmol/L ammonium formiate, were chosen. The mass
spectrometer used for this experiments was a Thermo Q-exactive.
For APPI a complex spectrum was obtained with a wide variety of signals, which can only
partly be assigned to TSP-ethoxylates like the signal of TSP ethoxylate with 33 EO units.
Given that the distribution of TSP-40 has its center on TSP with 33 EO units and not 40 EO
units and taking into account the scatter of smaller peaks underlying the distribution it may be
assumed that APPI is limited to ionization of entities with shorter EO chains. The ionization
process, however, of entities with longer chain length leads to some sort of degradation
shifting the center of distribution of ethoxylates and giving a wide variety of mass peaks,
being fragments of this process. As the spectrum is very hard to interpret APPI is less
favorable for characterization of TSP ethoxylates with longer EO chain lengths.
By comparison, the spectrum obtained by ESI shows only single to fourfold-charged mol
peaks of TSP ethoxylates without apparent degradation products or fragments. Analogous to
the spectrum obtained for TSP-16 in the manuscript in Figure 3 (b) the higher charged entities
11
are dominant for longer EO chain lengths. The spectrum obtained by ESI was easier to
interpret and without apparent degradation products and so ESI was taken as coupling to the
mass spectrometer in this work.
4. Determination of the limit of quantification
The limits of quantification (LOQ) for both analytes hexanophenone and TSP with 16 EO
units has been defined as a signal-to-noise ratio of at least 20:1 to ensure acceptable
quantification results. In the following the respective chromatograms at LOQ level are given
for TSP with 16 EO units (a) and hexanophenone (b) in Figure S 3, the linearity plots for TSP
with 16 EO units (a) and hexanophenone (b) in Figure S 4 and the distribution of residues for
TSP with 16 EO units (a) and hexanophenone (b) in Figure S 5. The data for the linear
regression were found to be heteroscedastic according to the Breusch-Pagan with the results
Signal intensity [counts]
shown in Table S 2.
Acquisition time [min]
(a)
12
Signal intensity [counts]
Acquisition time [min]
(b)
Figure S 3: Chromatograms for determination of the signal-to-noise ratio at the defined LOQ level for
TSP with 16 EO units (a) and hexanophenone (b). The LOQ was defined as a signal-to-noise ratio of at
least 20:1, which has been achieved for both analytes.
12000000
10000000
counts
8000000
6000000
y = 311388x + 73462
R² = 0.9998
4000000
2000000
0
0
5
10
15
20
25
30
35
40
c [mg/L]
(a)
13
3000000
2500000
counts
2000000
1500000
1000000
y = 22835x - 26727
R² = 0.9996
500000
0
0
20
40
60
80
100
120
140
c [mg/L]
(b)
Figure S 4: Linear ranges for TSP with 16 EO units (a) and hexanophenone (b) including the bands of
prediction indicated green for the upper and red for the lower limit
Standardisierte Residuen
2.5
2
1.5
1
0.5
0
-0.5
0
5
10
15
20
25
30
35
-1
-1.5
Content TSP with 16-EO units [mg/L]
a)
14
Standardisierte Residuen
1
0.5
0
0
20
40
60
80
100
120
140
-0.5
-1
-1.5
-2
Content ISTD [mg/L]
b)
Figure S 5: Distribution of residues for the regression analysis of standards of TSP with 16 EO units a)
and the internal standard hexanophenone b)
Table S 2: Results for the test on heteroscedasticity against a level of significance of 0.05 on the data set
used for linear regression of TSP-with 16 EO units and the internal standard hexanophenone according to
Breusch-Pagan
TSP with 16 EO units
Hexanophenone (ISTD)
p-Value
0.004
0.004
Hypothesis
H0 (Residues are
H0 (Residues are
homoscedastic) rejected
homoscedastic) rejected
Although the residues for the linear regressions were heteroscedastic, this was not considered
for the calculation of the linear regression. As the aim of this work is the relative comparison
of different suppliers of TSP-16 and not an absolute quantification of the single components
this limitation is acceptable.
15
5. Data bases and results files for data mining
The compound search applied in this work, is a targeted on known compounds. The
corresponding database is added as .csv file to the Supporting Information (TSP exact mass
data base.csv). The resulting data set for the 2-step data mining approach on the nonionic
surfactants in all the analyzed samples is added as a .txt file (TSP-16-ethoxylates_final.txt) to
the Supporting Information together with the intermediate data sets obtained during data
mining (TSP-16-ethoxylates molecular feature extractor.txt), normalization and reduction
(TSP-16-ethoxylates Find-by-Formular.txt) to the defined linear range. The resulting data set
on TSP 16 and TSP-16 in formulation samples is also added here (TSP-16-ethoxylates plus
formulation samples.txt).
6. Results of the principal components analysis according to the score of the
single compounds
In the following Table S 3 are shown the single scores each compound for component 1 and 2
of the principle component analysis as shown graphically in Figure 6 in the manuscript.
Table S 3: Given are the scores each compound for component 1 and 2 of the principle component analysis
performed on the data set of the different suppliers of TSP-16, as it is graphically displayed in Figure 6 in
the manuscript.
Compound
Hexanophenon
Monostyryphenolethoxylate-11-EO
Monostyryphenolethoxylate-12-EO
Monostyryphenolethoxylate-13-EO
Monostyryphenolethoxylate-14-EO
Monostyryphenolethoxylate-15-EO
Monostyryphenolethoxylate-16-EO
Monostyryphenolethoxylate-17-EO
Monostyryphenolethoxylate-18-EO
Monostyryphenolethoxylate-19-EO
Component 1
0
-21.518444
-17.013428
-20.885107
-25.291218
-32.93415
-25.511671
-24.716763
-24.73755
-26.527723
Component 2
0
18.958761
17.149254
20.679117
23.80306
28.694887
39.14467
39.249256
38.80539
40.04413
Retention
Mass
Time
176.1201
4.58
682.392
4.92
726.4198
4.94
770.4452
4.96
814.4709
4.99
858.4977
5.01
902.5234
5.03
946.5495
5.05
990.5766
5.07
1034.602
5.08
16
Compound
Monostyryphenolethoxylate-20-EO
Monostyryphenolethoxylate-21-EO
Monostyryphenolethoxylate-22-EO
Monostyryphenolethoxylate-23-EO
Monostyryphenolethoxylate-24-EO
Distyryphenolethoxylate-5-EO
Distyryphenolethoxylate-6-EO
Distyryphenolethoxylate-7-EO
Distyryphenolethoxylate-8-EO
Distyryphenolethoxylate-9-EO
Distyryphenolethoxylate-10-EO
Distyryphenolethoxylate-11-EO
Distyryphenolethoxylate-12-EO
Distyryphenolethoxylate-13-EO
Distyryphenolethoxylate-14-EO
Distyryphenolethoxylate-15-EO
Distyryphenolethoxylate-16-EO
Distyryphenolethoxylate-17-EO
Distyryphenolethoxylate-18-EO
Distyryphenolethoxylate-19-EO
Distyryphenolethoxylate-20-EO
Distyryphenolethoxylate-21-EO
Distyryphenolethoxylate-22-EO
Distyryphenolethoxylate-23-EO
Distyryphenolethoxylate-24-EO
Distyryphenolethoxylate-25-EO
Distyryphenolethoxylate-26-EO
Distyryphenolethoxylate-27-EO
Distyryphenolethoxylate-28-EO
Distyryphenolethoxylate-29-EO
Tristyryphenolethoxylate-5-EO
Tristyryphenolethoxylate-6-EO
Tristyryphenolethoxylate-7-EO
Tristyryphenolethoxylate-8-EO
Tristyryphenolethoxylate-9-EO
Tristyryphenolethoxylate-10-EO
Tristyryphenolethoxylate-11-EO
Tristyryphenolethoxylate-12-EO
Tristyryphenolethoxylate-13-EO
Tristyryphenolethoxylate-14-EO
Tristyryphenolethoxylate-15-EO
Tristyryphenolethoxylate-16-EO
Tristyryphenolethoxylate-17-EO
Tristyryphenolethoxylate-18-EO
Tristyryphenolethoxylate-19-EO
Tristyryphenolethoxylate-20-EO
Component 1
Component 2
-15.867841
32.809223
-19.358452
27.466583
-18.085913
16.839708
-15.225504
13.234964
-10.874819
8.432772
-2.6254199
5.172396
-4.185789
2.4134681
-0.9710141
2.3733385
-1.0998861
3.294223
-2.7358875
1.6297657
-1.941136
1.4260204
-1.9511255
1.3822399
-1.9803915
1.2908698
-1.9467528
1.2308027
-1.8844413
1.1279616
-1.8335105
1.0220265
-1.718206
0.90500146
-1.6626208
0.81457263
-1.5110463
0.68221706
-1.4079044
0.5977382
-1.2562836
0.45425314
-1.150982
0.40137506
-0.9551027
0.27022606
0.3792616
-3.9362864
-0.6929657
0.028805576
-4.2088923
7.1047454
-1.7542683
-0.26916566
0.5838643
4.580197
-6.2823634
-3.9688544
25.656946
-21.225237
-3.4106627
1.4796438
-1.2727805
1.1529844
-1.2660158
1.2475859
-0.28587636
0.8669842
-0.3896259
0.77770567
-0.43556568
0.61594385
-0.47407123
0.57370925
-0.4584877
0.45355675
-0.41510287
0.36597556
-0.36435264
0.29770425
-0.27751935
0.1869533
-0.193165
0.05831029
-0.10277939
-0.066116735
0.025035297
-0.1622553
0.14945453
-0.3214445
0.26133797
-0.3555113
Retention
Mass
Time
1078.629
5.10
1122.653
5.11
1166.68
5.13
1210.703
5.14
1254.729
5.16
522.3015
5.53
566.3277
5.55
610.3523
5.56
654.3778
5.58
698.4035
5.59
742.4296
5.60
786.4559
5.60
830.4822
5.61
874.5083
5.62
918.5346
5.63
962.5605
5.64
1006.587
5.64
1050.613
5.65
1094.639
5.66
1138.665
5.66
1182.692
5.67
1226.718
5.67
1270.744
5.68
1314.77
5.68
1358.796
5.68
1402.823
5.68
1446.849
5.69
1490.875
5.69
1534.901
5.69
1578.929
5.65
626.361
6.00
670.3875
6.01
714.4137
6.01
758.4396
6.01
802.4657
6.02
846.492
6.02
890.5182
6.02
934.5444
6.02
978.5708
6.02
1022.597
6.03
1066.623
6.03
1110.649
6.03
1154.675
6.03
1198.702
6.03
1242.728
6.03
1286.754
6.03
17
Compound
Tristyryphenolethoxylate-21-EO
Tristyryphenolethoxylate-22-EO
Tristyryphenolethoxylate-23-EO
Tristyryphenolethoxylate-24-EO
Tristyryphenolethoxylate-25-EO
Tristyryphenolethoxylate-26-EO
Tristyryphenolethoxylate-27-EO
Tristyryphenolethoxylate-28-EO
Tristyryphenolethoxylate-29-EO
Tristyryphenolethoxylate-30-EO
Tristyryphenolethoxylate-31-EO
Tetrastyryphenolethoxylate-5-EO
Tetrastyryphenolethoxylate-6-EO
Tetrastyryphenolethoxylate-7-EO
Tetrastyryphenolethoxylate-8-EO
Tetrastyryphenolethoxylate-9-EO
Tetrastyryphenolethoxylate-10-EO
Tetrastyryphenolethoxylate-11-EO
Tetrastyryphenolethoxylate-12-EO
Tetrastyryphenolethoxylate-13-EO
Tetrastyryphenolethoxylate-14-EO
Tetrastyryphenolethoxylate-15-EO
Tetrastyryphenolethoxylate-16-EO
Tetrastyryphenolethoxylate-17-EO
Tetrastyryphenolethoxylate-18-EO
Tetrastyryphenolethoxylate-19-EO
Tetrastyryphenolethoxylate-20-EO
Tetrastyryphenolethoxylate-21-EO
Tetrastyryphenolethoxylate-22-EO
Tetrastyryphenolethoxylate-23-EO
Tetrastyryphenolethoxylate-24-EO
Tetrastyryphenolethoxylate-25-EO
Monostyrylphenolprop-ethoxylate5-EO-8-PO
Monostyrylphenolprop-ethoxylate6-EO-8-PO
Monostyrylphenolprop-ethoxylate7-EO-8-PO
Monostyrylphenolprop-ethoxylate8-EO-8-PO
Monostyrylphenolprop-ethoxylate9-EO-8-PO
Monostyrylphenolprop-ethoxylate10-EO-8-PO
Monostyrylphenolprop-ethoxylate11-EO-8-PO
Monostyrylphenolprop-ethoxylate12-EO-8-PO
Component 1
Component 2
0.42371926
-0.5586756
0.46660414
-0.41179907
0.70624554
-0.7611214
0.3835046
-0.1521237
-0.5297137
-1.205153
-0.9155471
0.63756394
-4.92467
-0.46643406
-2.411118
1.9084175
-7.3978014
-11.465272
40.23051
-31.489828
13.274889
0.54010266
-3.7890558
-8.516457
-12.51962
-19.076212
-4.578877
-27.566341
-19.086279
-12.592126
-6.1373577
-26.144741
-13.65941
-10.564696
-6.8554664
-3.5307236
-7.446675
-6.3279986
-3.7851732
-4.298947
-1.3549933
-0.73320895
-1.3206834
-0.87198585
-2.1694055
-0.7420312
-6.312905
-8.372017
-4.6640363
-1.1875252
-15.150156
-6.937315
-7.81072
-5.7653356
-14.134495
-24.12576
-14.034096
-10.08268
-17.998505
-12.262659
13.857056
-18.175201
0.31084424
-0.30325606
Retention
Mass
Time
1330.78
6.03
1374.807
6.03
1418.833
6.03
1462.859
6.02
1506.885
6.03
1550.911
6.02
1594.937
6.03
1638.963
6.02
1682.99
6.02
1727.015
6.00
1771.045
6.06
730.4228
6.31
774.4505
6.33
818.4767
6.36
862.5024
6.37
906.5262
6.35
950.5555
6.38
994.5801
6.38
1038.607
6.38
1082.633
6.38
1126.66
6.38
1170.686
6.38
1214.712
6.38
1258.738
6.37
1302.764
6.37
1346.79
6.37
1390.817
6.37
1434.843
6.35
1478.87
6.36
1522.895
6.34
1566.919
6.35
1610.946
6.39
26.74786
13.669259 882.5688
6.09
17.234734
8.269603 926.5933
6.12
29.101526
14.494107 970.6096
6.08
33.514618
16.18774 1014.637
6.06
35.42035
15.915241 1058.659
6.05
25.031956
16.850344 1102.685
6.06
17.60985
13.992486
1146.71
6.06
31.77927
15.785249 1190.735
6.05
18
Compound
Monostyrylphenolprop-ethoxylate13-EO-8-PO
Distyrylphenolprop-ethoxylate-5EO-8-PO
Distyrylphenolprop-ethoxylate-6EO-8-PO
Distyrylphenolprop-ethoxylate-7EO-8-PO
Distyrylphenolprop-ethoxylate-8EO-8-PO
Distyrylphenolprop-ethoxylate-9EO-8-PO
Distyrylphenolprop-ethoxylate-10EO-8-PO
Distyrylphenolprop-ethoxylate-11EO-8-PO
Distyrylphenolprop-ethoxylate-12EO-4-PO
Distyrylphenolprop-ethoxylate-12EO-8-PO
Distyrylphenolprop-ethoxylate-13EO-8-PO
Distyrylphenolprop-ethoxylate-14EO-8-PO
Distyrylphenolprop-ethoxylate-15EO-8-PO
Distyrylphenolprop-ethoxylate-16EO-8-PO
Distyrylphenolprop-ethoxylate-17EO-8-PO
Distyrylphenolprop-ethoxylate-18EO-8-PO
Distyrylphenolprop-ethoxylate-19EO-8-PO
Component 1
Component 2
Mass
Retention
Time
5.6304994
0.3463299 1234.761
6.04
57.404755
7.765355 986.6327
6.39
57.82702
7.887388
1030.66
6.38
58.48031
7.788493 1074.686
6.38
58.59349
7.7376966 1118.711
6.38
58.646797
7.669096 1162.738
6.38
58.14472
7.6415954 1206.764
6.38
57.51143
7.6519156 1250.789
6.37
1.0730832
0.5492888 1062.638
6.05
56.77911
7.385921 1294.816
6.37
54.253757
9.28449 1338.839
6.37
54.626804
7.914997 1382.868
6.37
37.83858
4.5463367 1426.891
6.35
49.122604
8.906896 1470.919
6.37
6.9875383
-1.7860774 1514.936
6.29
37.346233
2.8904805 1558.969
6.35
3.4635794
0.73942816 1602.998
6.34
7. Comparison of TSP-16 of different suppliers and qualities
In Table S 4 the compounds in the different arrays determined by the hierarchical clustering
(HCA) performed on the combined supplier data set are given.
Table S 4: Compounds used for the combined hierarchical clustering listed together with the
corresponding arrays as defined in Figure 7 (see manuscript). The compounds are sorted according to the
order obtained by the hierarchical clustering of the compounds.
Array
1
Compound
Distyrylphenolprop-ethoxylate-16-
Array
5
Compound
Distyrylphenolethoxylate-15-EO
19
Array
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
2
2
Compound
EO 8-PO
Distyrylphenolprop-ethoxylate-13EO 8-PO
Array
Compound
5
Distyrylphenolethoxylate-18-EO
Distyrylphenolprop-ethoxylate-11EO 8-PO
Distyrylphenolprop-ethoxylate-12EO 8-PO
Distyrylphenolprop-ethoxylate-14EO 8-PO
Distyrylphenolprop-ethoxylate-8-EO
8-PO
Distyrylphenolprop-ethoxylate-7-EO
8-PO
Distyrylphenolprop-ethoxylate-9-EO
8-PO
Distyrylphenolprop-ethoxylate-5-EO
8-PO
Distyrylphenolprop-ethoxylate-6-EO
8-PO
Distyrylphenolprop-ethoxylate-10EO 8-PO
Monostyrylphenolprop-ethoxylate11-EO 8-PO
Monostyrylphenolprop-ethoxylate8-EO 8-PO
Monostyrylphenolprop-ethoxylate9-EO 8-PO
Monostyrylphenolprop-ethoxylate5-EO 8-PO
Monostyrylphenolprop-ethoxylate12-EO 8-PO
Monostyrylphenolprop-ethoxylate7-EO 8-PO
5
Tristyrylphenolethoxylate-22-EO
5
Tristyrylphenolethoxylate-21-EO
5
Tristyrylphenolethoxylate-11-EO
5
Tristyrylphenolethoxylate-24-EO
5
Tristyrylphenolethoxylate-10-EO
5
Distyrylphenolethoxylate-21-EO
5
Distyrylphenolethoxylate-12-EO
5
Distyrylphenolethoxylate-20-EO
5
Tristyrylphenolethoxylate-9-EO
5
Tristyrylphenolethoxylate-23-EO
5
Tristyrylphenolethoxylate-8-EO
5
Hexanophenone (Internal Standard)
5
Distyrylphenolethoxylate-22-EO
5
Distyrylphenolethoxylate-11-EO
5
Distyrylphenolethoxylate-10-EO
Monostyrylphenolprop-ethoxylate10-EO 8-PO
Monostyrylphenolprop-ethoxylate6-EO 8-PO
Distyrylphenolprop-ethoxylate-15EO 8-PO
Distyrylphenolprop-ethoxylate-18EO 8-PO
Tristyrylphenolethoxylate-31-EO
Distyrylphenolprop-ethoxylate-19EO 8-PO
5
Tristyrylphenolethoxylate-26-EO
5
Tristyrylphenolethoxylate-16-EO
5
Tristyrylphenolethoxylate-15-EO
5
Tristyrylphenolethoxylate-17-EO
5
5
Tristyrylphenolethoxylate-18-EO
Tristyrylphenolethoxylate-14-EO
20
Array
2
2
2
2
2
2
2
2
2
3
3
3
3
3
3
3
3
3
3
3
3
3
4
4
4
4
4
4
4
5
5
5
5
5
Compound
Distyrylphenolprop-ethoxylate-12EO 8-PO
Tetrastyrylphenolethoxylate-25-EO
Array
5
Compound
Tristyrylphenolethoxylate-19-EO
5
Tristyrylphenolethoxylate-13-EO
Monostyrylphenolprop-ethoxylate13-EO 8-PO
Tetratstyrylphenolethoxylate-5-EO
Distyrylphenolprop-ethoxylate-17EO 8-PO
Monostyrylphenolethoxylate-24-EO
Tetrastyrylphenolethoxylate-6-EO
Tetrastyrylphenolethoxylate-24-EO
Distyrylphenolethoxylate-29-EO
Monostyrylphenolethoxylate-18-EO
Monostyrylphenolethoxylate-19-EO
Monostyrylphenolethoxylate-16-EO
Monostyrylphenolethoxylate-17-EO
Monostyrylphenolethoxylate-15-EO
Monostyrylphenolethoxylate-21-EO
Monostyrylphenolethoxylate-13-EO
Monostyrylphenolethoxylate-14-EO
Monostyrylphenolethoxylate-12-EO
Monostyrylphenolethoxylate-20-EO
Monostyrylphenolethoxylate-11-EO
Monostyrylphenolethoxylate-23-EO
Monostyrylphenolethoxylate-22-EO
Tristyrylphenolethoxylate-29-EO
Tetrastyrylphenolethoxylate-21-EO
Tetrastyrylphenolethoxylate-23-EO
Tetrastyrylphenolethoxylate-8-EO
Tetrastyrylphenolethoxylate-7-EO
Tetrastyrylphenolethoxylate-9-EO
Tristyrylphenolethoxylate-30-EO
Distyrylphenolethoxylate-13-EO
Distyrylphenolethoxylate-19-EO
Distyrylphenolethoxylate-14-EO
Distyrylphenolethoxylate-17-EO
Distyrylphenolethoxylate-16-EO
5
Tristyrylphenolethoxylate-20-EO
5
5
Tristyrylphenolethoxylate-12-EO
Tristyrylphenolethoxylate-28-EO
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
5
Tetrastyrylphenolethoxylate-14-EO
Tetrastyrylphenolethoxylate-15-EO
Distyrylphenolethoxylate-24-EO
Tetrastyrylphenolethoxylate-16-EO
Tristyrylphenolethoxylate-6-EO
Distyrylphenolethoxylate-7-EO
Distyrylphenolethoxylate-8-EO
Tristyrylphenolethoxylate-7-EO
Distyrylphenolethoxylate-9-EO
Tristyrylphenolethoxylate-25-EO
Distyrylphenolethoxylate-23-EO
Tristyrylphenolethoxylate-5-EO
Tetrastyrylphenolethoxylate-18-EO
Tetrastyrylphenolethoxylate-13-EO
Tetrastyrylphenolethoxylate-12-EO
Tetrastyrylphenolethoxylate-17-EO
Distyrylphenolethoxylate-27-EO
Distyrylphenolethoxylate-5-EO
Distyrylphenolethoxylate-6-EO
Distyrylphenolethoxylate-26-EO
Tetrastyrylphenolethoxylate-22-EO
Tetrastyrylphenolethoxylate-20-EO
Tetrastyrylphenolethoxylate-11-EO
Tetrastyrylphenolethoxylate-19-EO
Tetrastyrylphenolethoxylate-10-EO
Tristyrylphenolethoxylate-27-EO
Distyrylphenolethoxylate-28-EO
Distyrylphenolethoxylate-25-EO
21
8. Statistical evaluation of the results on the content of the components in
TSP-16 on their use for product identification
As described, there were substantial differences in the contents of the main components,
MSP-, DSP-, TSP- and TeSP ethoxylates, and the by-products, copolymerized propoxylatesethoxylates of MSP and DSP, in the commercial TSP-16. These differences could be used as
signature for product identification. Therefore, model formulation samples were prepared
containing TSP-16 of each supplier and quality (A, B1, B2, C). In order to validate the
clustering on TSP-16 according to its suppliers, the samples used for the model formulation
were not part of the original data set. These samples were then analyzed in order to test if they
were assigned correctly to their suppliers using both PCA and hierarchical clustering. In the
following, the results of the PCA (Figure S 6) and of the HCA (Figure S 7) are displayed. In
case of the HCA the clustering according to the compounds (y-axis) is compressed as only the
Component 2 (22.1 %)
clustering according to the samples (x-axis) was of interest.
Component 1 (63.2 %)
Figure S 6: Principle component analysis of the data sets from supplier A (Cross), B1 (Arrow), B2
(Horizontal Bar) and C (Vertical bar) together with the data of the formulation samples containing TSP16 of supplier A (Square), B1 (Diamond), B2 (Circle) and C (Triangle). For the PCA the whole data set
was taken including the 3 repetition analysis each production batch and formulation sample.
22
Figure S 7: Combined hierarchical clustering of the samples (x-axis) and the compounds (y-axis) detected
in the samples of supplier A, B1, B2 and C together with sample of formulation containing TSP-16 of
Supplier A, B1, B2 and C. Each sample is the average of 3 repetition analyses. The content of a compound
in the analyzed sample is coded via black-white rectangles in the column beneath the respective samples.
The color ranges from black, compound not detected, over grey, compound as abundant as internal
standard, to white, compound with the maximum content.
As demonstrated, both data analysis techniques were correctly assigning the TSP-16 in the
model formulation to their corresponding suppliers. Using the PCA all four formulation
samples were identified in their corresponding supplier cluster. They were all group within the
clusters of their suppliers, as shown in the top dendrogram. The linkage of the formulation
samples to a sample of the corresponding supplier cluster in the HCA was formed for all four
samples at least two levels lower than the linkage of the respective supplier cluster. The
assignments to the corresponding suppliers displayed in Figure S 7 were thus reasonable. The
developed method combining instrumental analysis and multivariate data mining enables the
identification of a supplier of TSP-16, without apparent matrix interference even though
23
another functionalized PEG, an ethoxylated alcohol, had been used as well in the chosen
model formulation.
9. Example for interference on analysis of TSP-16 in agrochemical
formulations
The identification of the different suppliers in an agrochemical formulation can be interfered
by end group sulfated or phosphated TSP ethoxylates, if they are contained in the
agrochemical formulation. The chromatograms of commercially available TSP-16 terminal
phosphated (a) and sulfated (b) are shown in Figure S 8 obtained in the positive ionization
Signal intensity [counts]
mode with the identified entities of DSP-, TSP- and TeSP ethoxylates.
Acquisition time [min]
(a)
24
Signal intensity [counts]
Acquisition time [min]
(b)
Figure S 8: Extracted ion chromatograms obtained in the positive ionization mode of terminal phosphated
(a) and sulfated (b) commercially available TSP-16. Indicated are the identified entities of DSP-, TSP and
TeSP ethoxylates.
As shown there are entities of DSP-, TSP- and TeSP ethoxylates detectable in commercially
available terminal sulfated and phosphated TSP-16 in the chosen ionization mode. These
entities of DSP-, TSP- and TeSP ethoxylates can be explained by incomplete phosphating or
sulfating of the educt TSP-16 which was not removed after the reaction from the final
commercially product. Possible interferences of these entities on the identification of the
different suppliers of TSP-16 in the matrix of the model agrochemical formulations were
investigated next. TSP-16 of supplier A, B2 and C and terminal sulfated TSP-16 were mixed
in the model agrochemical as shown in Table S 5.
Table S 5: Table of composition of the model agrochemical formulation containing terminal sulfated TSP16 alongside with TSP-16
Raw material
Content [%] (w/w)
Active ingredient
23.0
TSP-16
2.5
25
TSP-16, sulfated
2.5
Dispersing agent (non-ionic)
10.0
Emulsifier 1 (non-ionic, functionalized PEG)
15.0
Emulsifier 2 (non-ionic, functionalized PPG-PEG-co-polymer)
9.0
Hydrophbically modified Clay
0.1
Acid
0.4
Solvent
37.5
These formulation samples were subjected to the analysis and multivariate data analysis
techniques developed and used in this work, with the results of the principal component
Component 2 (33.1
%)
analysis (PCA) shown in Figure S 9.
Component 1 (45.2 %)
Figure S 9: Principal component analysis of the data sets from supplier A (Cross), B1 (Arrow), B2
(Horizontal Bar) and C (Vertical bar) together with the data of the formulation samples containing TSP16 of supplier A (Square), B2 (Circle) and C (Triangle). For the PCA the whole data set was taken
including the 3 repetition analysis each production batch and formulation sample.
26
As shown the entities of DSP-, TSP- and TeSP ethoxylates contained in end group sulfated
TSP-16 interfere with the developed method. As formulations using a combination of TSP-16
and another TSP ethoxylates derivate are not widely spread this potential interference can be
accepted. Nevertheless, further investigations should test the possibility for a correction of the
observed interferences.
27