Determination of halogenated volatile organic …10.1007... · Web view1 University Duisburg-Essen, Instrumental Analytical Chemistry, Universitätsstr. 5, 45141 Essen, Germany 2

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: [email protected]

Table of Contents

1. Sample for testing on mass calibration of TOF-MS.................................................................................6

2. Mass spectra of the major compounds in TSP-16...................................................................................8

3. Comparison of the ionization performance of APPI and ESI for the analysis of TSP-40-ethoxylates.......11

4. Determination of the limit of quantification........................................................................................13

5. Data bases and results files for the data mining...................................................................................17

6. Results of the principal components analysis according to the score of the single compounds.............17

7. Comparison of TSP-16 of different suppliers and qualities....................................................................20

8. Statistical evaluation of the results on the content of the components in TSP-16 on their use for product identification............................................................................................................................................... 23

9. Example for interference on analysis of TSP-16 in agrochemical formulations......................................25

1

mailto:[email protected]

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 Q-

exactive.....................................................................................................................................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 signal-

to-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

2

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......................................................................................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...........................................................................................................................6

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............................................................................16

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.................................................................17

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....................20

Table S 5: Table of composition of the model agrochemical formulation containing terminal

sulfated TSP-16 alongside with TSP-16...................................................................................26

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 tN [min] Exact mass [m/z]Imidacloprid 2.0 254.0450Thiacloprid 2.5 252.0236Tebuconazole (1.Isomer) 4.3 307.1451Triadimenol 4.6 295.1088Tebuconazole (2.Isomer) 4.9 307.1451Distyrylethoxylate-5-EO 5.8 522.2981Distyrylethoxylate-6-EO 5.8 566.3244Distyrylethoxylate-7-EO 5.8 610.3506Distyrylethoxylate-8-EO 5.8 654.3768Distyrylethoxylate-9-EO 5.8 698.4030Distyrylethoxylate-10-EO 5.8 742.4292Distyrylethoxylate-11-EO 5.8 786.4554Distyrylethoxylate-12-EO 5.8 830.4816Distyrylethoxylate-13-EO 5.8 874.5079Distyrylethoxylate-14-EO 5.8 918.5341Distyrylethoxylate-15-EO 5.8 962.5603Distyrylethoxylate-16-EO 5.8 1006.5865Distyrylethoxylate-17-EO 5.9 1050.6127Distyrylethoxylate-18-EO 5.9 1094.6389Distyrylethoxylate-19-EO 5.9 1138.6651Distyrylethoxylate-20-EO 5.9 1182.6914Distyrylethoxylate-21-EO 5.9 1226.7176Distyrylethoxylate-22-EO 5.9 1270.7438Distyrylethoxylate-23-EO 5.9 1314.7700Distyrylethoxylate-24-EO 5.9 1358.7962Distyrylethoxylate-25-EO 5.9 1402.8224Distyrylethoxylate-26-EO 5.9 1446.8486Distyrylethoxylate-27-EO 5.9 1490.8749Distyrylethoxylate-28-EO 5.9 1534.9011Distyrylethoxylate-29-EO 5.9 1578.9273Distyrylethoxylate-30-EO 5.9 1622.9535Nonylphenolethoxylate-5-EO 6.6 440.3138Nonylphenolethoxylate-6-EO 6.3 484.3400Nonylphenolethoxylate-7-EO 6.2 528.3662Nonylphenolethoxylate-8-EO 6.2 572.3924Nonylphenolethoxylate-9-EO 6.2 616.4186Nonylphenolethoxylate-10-EO 6.2 660.4449Nonylphenolethoxylate-11-EO 6.2 704.4711Nonylphenolethoxylate-12-EO 6.2 748.4973Nonylphenolethoxylate-13-EO 6.2 792.5235Nonylphenolethoxylate-14-EO 6.2 836.5497Nonylphenolethoxylate-15-EO 6.2 880.5759

5

Compound tN [min] Exact mass [m/z]Nonylphenolethoxylate-16-EO 6.2 924.6022Nonylphenolethoxylate-17-EO 6.2 968.6284Nonylphenolethoxylate-18-EO 6.2 1012.6546Nonylphenolethoxylate-19-EO 6.2 1056.6808Nonylphenolethoxylate-20-EO 6.2 1100.7070Nonylphenolethoxylate-21-EO 6.2 1144.7332Nonylphenolethoxylate-22-EO 6.2 1188.7594Nonylphenolethoxylate-23-EO 6.2 1232.7857Nonylphenolethoxylate-24-EO 6.2 1276.8119Nonylphenolethoxylate-25-EO 6.2 1320.8381Nonylphenolethoxylate-26-EO 5.9 1364.8643Nonylphenolethoxylate-27-EO 5.9 1408.8905Nonylphenolethoxylate-28-EO 5.9 1452.9167Nonylphenolethoxylate-29-EO 5.9 1496.9429Nonylphenolethoxylate-30-EO 5.9 1540.9692Tristyrylethoxylate-5-EO 5.9 626.3607Tristyrylethoxylate-6-EO 5.9 670.38695Tristyrylethoxylate-7-EO 5.9 714.4132Tristyrylethoxylate-8-EO 6.5 758.4394Tristyrylethoxylate-9-EO 5.9 802.4656Tristyrylethoxylate-10-EO 5.9 846.4918Tristyrylethoxylate-11-EO 6.0 890.5180Tristyrylethoxylate-12-EO 6.0 934.5442Tristyrylethoxylate-13-EO 6.0 978.5705Tristyrylethoxylate-14-EO 6.0 1022.5967Tristyrylethoxylate-15-EO 6.0 1066.6229Tristyrylethoxylate-16-EO 6.0 1110.6491Tristyrylethoxylate-17-EO 6.0 1154.6753Tristyrylethoxylate-18-EO 6.0 1198.7015Tristyrylethoxylate-19-EO 6.0 1242.7278Tristyrylethoxylate-20-EO 6.0 1286.7540Tristyrylethoxylate-21-EO 5.9 1330.7802Tristyrylethoxylate-22-EO 5.9 1374.8064Tristyrylethoxylate-23-EO 5.9 1418.8326Tristyrylethoxylate-24-EO 5.9 1462.8588Tristyrylethoxylate-25-EO 5.9 1506.8850Tristyrylethoxylate-26-EO 5.8 1550.9113Tristyrylethoxylate-27-EO 5.8 1594.9375Tristyrylethoxylate-28-EO 5.8 1638.9637Tristyrylethoxylate-29-EO 5.8 1682.9899Tristyrylethoxylate-30-EO 5.8 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

(a)

Signal intensity [counts]

Mass-to-Charge [m/z]

PEG with 9 EO-units [M + NH4]+

PEG

7

(b)

(c)



MSP with 16 EO-units [M + NH4]+


(1) MSP-ethoxylates



DSP with 15 EO-units [M + NH4]+


(2) DSP-ethoxylates

8

(d)

(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



(3) TSP-ethoxylates

TSP with 15 EO-units [M + NH4]+PEG with 4 EO-units [M + NH4]+(3) TSP-ethoxylates



TeSP with 13 EO-units [M + NH4]+


(4) TeSP-ethoxylates

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-40-

ethoxylates. 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).

(a)Mass-to-Charge [m/z]

Relative abundance [%]

TSP with 33 EO-units [M + Na]+

10

(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


Relative abundance [%]

TSP with 43 EO-units [M + 4NH4]4+



TSP with 35 EO-units [M + NH4]+

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

shown in Table S 2.

(a)

Acquisition time [min]


12

(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.

0 5 10 15 20 25 30 35 400

2000000

4000000

6000000

8000000

10000000

12000000

f(x) = 311388.338794674 x + 73462.351121488R² = 0.999770749113375

c [mg/L]

coun

ts

(a)



13

0 20 40 60 80 100 120 1400

500000

1000000

1500000

2000000

2500000

3000000

f(x) = 22834.7144766107 x − 26727.4253717642R² = 0.999600645884538

c [mg/L]

coun

ts

(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

0 5 10 15 20 25 30 35

-1.5

-1

-0.5

0

0.5

1

1.5

2

2.5

Content TSP with 16-EO units [mg/L]

Stan

dard

isie

rte

Resi

duen

a)

14

0 20 40 60 80 100 120 140

-2

-1.5

-1

-0.5

0

0.5

1

Content ISTD [mg/L]

Stan

dard

isie

rte

Resi

duen

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 0.004Hypothesis H0 (Residues are

homoscedastic) rejectedH0 (Residues are 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 Component 1 Component 2 MassRetention Time

Hexanophenon 0 0176.120

1 4.58Monostyryphenolethoxylate-11-EO -21.518444 18.958761 682.392 4.92

Monostyryphenolethoxylate-12-EO -17.013428 17.149254726.419

8 4.94


2 4.96


9 4.99


7 5.01

16



4 5.03


5 5.05


6 5.07


2 5.08


9 5.10


3 5.11Monostyryphenolethoxylate-22-EO -18.085913 16.839708 1166.68 5.13


3 5.14


9 5.16

Distyryphenolethoxylate-5-EO -2.6254199 5.172396522.301

5 5.53


7 5.55


3 5.56


8 5.58


5 5.59


6 5.60


9 5.60


2 5.61


3 5.62


6 5.63


5 5.64


7 5.64


3 5.65


9 5.66


5 5.66


2 5.67


8 5.67


4 5.68

17


Distyryphenolethoxylate-23-EO 0.3792616 -3.9362864 1314.77 5.68


6 5.68


3 5.68

Distyryphenolethoxylate-26-EO -1.7542683 -0.269165661446.84

9 5.69

Distyryphenolethoxylate-27-EO 0.5838643 4.5801971490.87

5 5.69

Distyryphenolethoxylate-28-EO -6.2823634 -3.96885441534.90

1 5.69

Distyryphenolethoxylate-29-EO 25.656946 -21.2252371578.92

9 5.65Tristyryphenolethoxylate-5-EO -3.4106627 1.4796438 626.361 6.00

Tristyryphenolethoxylate-6-EO -1.2727805 1.1529844670.387

5 6.01


7 6.01


6 6.01


7 6.02Tristyryphenolethoxylate-10-EO -0.43556568 0.61594385 846.492 6.02


2 6.02


4 6.02


8 6.02


7 6.03


3 6.03


9 6.03

Tristyryphenolethoxylate-17-EO -0.10277939 -0.0661167351154.67

5 6.03

Tristyryphenolethoxylate-18-EO 0.025035297 -0.16225531198.70

2 6.03


8 6.03


4 6.03Tristyryphenolethoxylate-21-EO 0.42371926 -0.5586756 1330.78 6.03


7 6.03


3 6.03


9 6.02


5 6.03

18



1 6.02


7 6.03


3 6.02Tristyryphenolethoxylate-29-EO -7.3978014 -11.465272 1682.99 6.02


5 6.00

Tristyryphenolethoxylate-31-EO 13.274889 0.540102661771.04

5 6.06

Tetrastyryphenolethoxylate-5-EO -3.7890558 -8.516457730.422

8 6.31


5 6.33


7 6.36


4 6.37


2 6.35


5 6.38


1 6.38


7 6.38


3 6.38Tetrastyryphenolethoxylate-14-EO -1.3549933 -0.73320895 1126.66 6.38


6 6.38


2 6.38


8 6.37




7 6.37




5 6.34

Tetrastyryphenolethoxylate-24-EO 13.857056 -18.1752011566.91

9 6.35

Tetrastyryphenolethoxylate-25-EO 0.31084424 -0.303256061610.94

6 6.39Monostyrylphenolprop-ethoxylate-5-EO-8-PO 26.74786 13.669259

882.5688 6.09

19


Monostyrylphenolprop-ethoxylate-6-EO-8-PO 17.234734 8.269603

926.5933 6.12


970.6096 6.08


1014.637 6.06


1058.659 6.05


1102.685 6.06

Monostyrylphenolprop-ethoxylate-11-EO-8-PO 17.60985 13.992486 1146.71 6.06Monostyrylphenolprop-ethoxylate-12-EO-8-PO 31.77927 15.785249

1190.735 6.05


1234.761 6.04

Distyrylphenolprop-ethoxylate-5-EO-8-PO

57.404755 7.765355986.632

7 6.39Distyrylphenolprop-ethoxylate-6-EO-8-PO 57.82702 7.887388 1030.66 6.38Distyrylphenolprop-ethoxylate-7-EO-8-PO 58.48031 7.788493

1074.686 6.38

Distyrylphenolprop-ethoxylate-8-EO-8-PO 58.59349 7.7376966

1118.711 6.38


1162.738 6.38


1206.764 6.38


1250.789 6.37


1062.638 6.05


1294.816 6.37


1338.839 6.37


1382.868 6.37


1426.891 6.35


1470.919 6.37

Distyrylphenolprop-ethoxylate-17-EO-8-PO 6.9875383 -1.7860774

1514.936 6.29


1558.969 6.35


1602.998 6.34

20

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 Compound Array Compound1 Distyrylphenolprop-ethoxylate-16-

EO 8-PO5 Distyrylphenolethoxylate-15-EO

1 Distyrylphenolprop-ethoxylate-13-EO 8-PO

5 Distyrylphenolethoxylate-18-EO


5 Tristyrylphenolethoxylate-22-EO

















1 Monostyrylphenolprop-ethoxylate-11-EO 8-PO





5 Hexanophenone (Internal Standard)







21

Array Compound Array Compound









2 Tristyrylphenolethoxylate-31-EO 5 Tristyrylphenolethoxylate-18-EO2 Distyrylphenolprop-ethoxylate-19-

EO 8-PO5 Tristyrylphenolethoxylate-14-EO



2 Tetrastyrylphenolethoxylate-25-EO 5 Tristyrylphenolethoxylate-13-EO



2 Tetratstyrylphenolethoxylate-5-EO 5 Tristyrylphenolethoxylate-12-EO2 Distyrylphenolprop-ethoxylate-17-

EO 8-PO5 Tristyrylphenolethoxylate-28-EO

2 Monostyrylphenolethoxylate-24-EO 5 Tetrastyrylphenolethoxylate-14-EO2 Tetrastyrylphenolethoxylate-6-EO 5 Tetrastyrylphenolethoxylate-15-EO2 Tetrastyrylphenolethoxylate-24-EO 5 Distyrylphenolethoxylate-24-EO2 Distyrylphenolethoxylate-29-EO 5 Tetrastyrylphenolethoxylate-16-EO3 Monostyrylphenolethoxylate-18-EO 5 Tristyrylphenolethoxylate-6-EO3 Monostyrylphenolethoxylate-19-EO 5 Distyrylphenolethoxylate-7-EO3 Monostyrylphenolethoxylate-16-EO 5 Distyrylphenolethoxylate-8-EO3 Monostyrylphenolethoxylate-17-EO 5 Tristyrylphenolethoxylate-7-EO3 Monostyrylphenolethoxylate-15-EO 5 Distyrylphenolethoxylate-9-EO3 Monostyrylphenolethoxylate-21-EO 5 Tristyrylphenolethoxylate-25-EO3 Monostyrylphenolethoxylate-13-EO 5 Distyrylphenolethoxylate-23-EO3 Monostyrylphenolethoxylate-14-EO 5 Tristyrylphenolethoxylate-5-EO3 Monostyrylphenolethoxylate-12-EO 5 Tetrastyrylphenolethoxylate-18-EO3 Monostyrylphenolethoxylate-20-EO 5 Tetrastyrylphenolethoxylate-13-EO3 Monostyrylphenolethoxylate-11-EO 5 Tetrastyrylphenolethoxylate-12-EO3 Monostyrylphenolethoxylate-23-EO 5 Tetrastyrylphenolethoxylate-17-EO3 Monostyrylphenolethoxylate-22-EO 5 Distyrylphenolethoxylate-27-EO4 Tristyrylphenolethoxylate-29-EO 5 Distyrylphenolethoxylate-5-EO4 Tetrastyrylphenolethoxylate-21-EO 5 Distyrylphenolethoxylate-6-EO4 Tetrastyrylphenolethoxylate-23-EO 5 Distyrylphenolethoxylate-26-EO4 Tetrastyrylphenolethoxylate-8-EO 5 Tetrastyrylphenolethoxylate-22-EO4 Tetrastyrylphenolethoxylate-7-EO 5 Tetrastyrylphenolethoxylate-20-EO4 Tetrastyrylphenolethoxylate-9-EO 5 Tetrastyrylphenolethoxylate-11-EO4 Tristyrylphenolethoxylate-30-EO 5 Tetrastyrylphenolethoxylate-19-EO5 Distyrylphenolethoxylate-13-EO 5 Tetrastyrylphenolethoxylate-10-EO

22

Array Compound Array Compound5 Distyrylphenolethoxylate-19-EO 5 Tristyrylphenolethoxylate-27-EO5 Distyrylphenolethoxylate-14-EO 5 Distyrylphenolethoxylate-28-EO5 Distyrylphenolethoxylate-17-EO 5 Distyrylphenolethoxylate-25-EO5 Distyrylphenolethoxylate-16-EO

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 propoxylates-

ethoxylates 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

clustering according to the samples (x-axis) was of interest.

23

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.

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

Component 1 (63.2 %)


24

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

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

mode with the identified entities of DSP-, TSP- and TeSP ethoxylates.

25

(a)

(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





26

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 TSP-

16 alongside with TSP-16

Raw material Content [%] (w/w)

Active ingredient 23.0

TSP-16 2.5

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

analysis (PCA) shown in Figure S 9.

27

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.

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.



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Determination of halogenated volatile organic …10.1007... · Web view1 University Duisburg-Essen, Instrumental Analytical Chemistry, Universitätsstr. 5, 45141 Essen, Germany 2