1

WAT E R S SO LU T IO NS

ACQUITY UPC2® System

Xevo® TQD

MassLynx® Software

ACQUITY UPC2 PDA Detector

Empower® 3 Software

K E Y W O R D S

Triton-X, cosmetics, personal care products,

household and industrial cleaning products

A P P L I C AT IO N B E N E F I T S

UPC2® with either UV or MS detection for the

analysis of non-ionic surfactant, offers:

■■ High-efficiency separation with excellent

resolution for approximately 20 oligomers.

■■ Analysis time less than 2 min

with PDA detection.

■■ Reduction in consumption

of organic solvents.

■■ Analysis at lower temperatures

than in GC or SFC.

■■ The detection of: additional minor series

components; by-products; impurities;

degradation products or contaminants.

IN T RO DU C T IO N

The non-ionic surfactant Triton X-100 (Figure 1), an excellent detergent and

wetting agent, is readily biodegradable and achieves effective performance across

a broad temperature range. It can also be used as a dispersant and emulsifier for

oil in water systems. Because of these properties, Triton X-100 is used in many

household and industrial cleaning products, paints and coatings, pulp and paper,

oil fields, textiles, agrochemicals, cosmetics, and industrial materials.

Analysis of the Non-Ionic Surfactant Triton-X Using UltraPerformance Convergence Chromatography (UPC2) with MS and UV Detection Jane Cooper,1 Baiba Cabovska2

1Waters Corporation, Wilmslow, UK2 Waters Corporation, Milford, MA, USA

(C14H22O(C2H4O)n) n= 9-10 Figure 1. Triton-X-100 structure

and chemical formula.

It is essential to be able to monitor the composition of the non-ionic, octylphenol

ethoxylate surfactant Triton X-100, because differences in the ethoxy chain length

can affect characteristics of the mixture such as viscosity, solubility, and polarity.

The ability to detect the presence of by-products, impurities, degradation

products or contaminants present in surfactants is equally important. In addition

to identifying potential carcinogenic or allergenic compounds, the presence of

impurities can also affect the efficiency of the surfactant.

Surfactants are typically analyzed using techniques such as High Performance

Liquid Chromatography (HPLC),1,2 Supercritical Fluid Chromatography (SFC),3 or

Gas Chromatography (GC).4,5 Analysis by GC and HPLC can be time consuming, as

these techniques may require additional derivatization stages in order to improve

sensitivity, separation or resolve volatilization issues. GC or traditional SFC

techniques that employ high column temperatures can also limit the analysis of

thermally labile compounds. In some cases, baseline separations for oligomers

using HPLC, SFC or GC analyses are not achieved.

2

E X P E R IM E N TA L

UV conditionsUV system: ACQUITY UPC2 PDA Detector

Range: 210 to 400 nm

Resolution: 4.8 nm

UPC2 System: ACQUITY UPC2

Column: ACQUITY UPC2 BEH 2.1 mm x 50 mm, 1.7 µm

Column temp.: 40 °C

Convergence column manager back pressure: 1500 psi

Injection volume: 1.0 µL

Mobile phase B: Methanol

Mobile phase gradient for UV detection is detailed in Table 1.

Time (min)

Flow rate (mL/min) %A %B Curve

1 Initial 2.00 98.0 2.0 —

2 1.25 2.00 65.0 35.0 6

3 1.30 2.00 98.0 2.0 6

4 2.00 2.00 98.0 2.0 6

Table 1. ACQUITY UPC 2 mobile phase gradient for UV detection.

Instrument control, data acquisition, and result processing

Empower 3 Software was used to control the ACQUITY UPC2

System and ACQUITY UPC2 PDA Detector, and provide data

acquisition and processing.

MassLynx Software was used to control the ACQUITY UPC2 System

and Xevo TQD, and provide data acquisition and processing.

Waters® UltraPerformance Convergence Chromatography™ (UPC2) System, builds on the potential of normal-phase separation techniques

such as SFC, while using proven Waters’ easy-to-use UPLC® Technology.

This application note describes the analysis Triton X-100 utilizing UPC2 with PDA and MS detection. Excellent resolution for approximately

20 oligomers has been achieved using lower temperatures than GC or traditional SFC analysis, making UPC2 more amenable for the analysis

of thermally labile compounds. A significant reduction in the consumption of toxic solvents was also achieved compared to normal phase

HPLC analysis.

MS conditions MS system: Xevo TQD

Ionization mode: ESI +

Capillary voltage: 3.5 kV

Source temp.: 150 °C

Desolvation temp.: 500 °C

Desolvation gas flow: 800 L/hr

Cone gas flow: 50 L/hr

Acquisition: Full scan

UPC2 System: ACQUITY UPC2

Column: ACQUITY UPC2 BEH 2.1 mm x 50 mm, 1.7 µm

Column temp.: 65 °C

CCM back pressure: 1600 psi

Injection volume: 1.0 µL

Mobile phase B: Methanol

Mobile phase gradient for MS detection is detailed in Table 2.

Time (min)

Flow rate (mL/min) %A %B Curve

1 Initial 2.00 97.0 3.0 –

2 20.00 2.00 80.0 20.0 6

3 21.00 2.00 97.0 3.0 6

4 23.00 2.00 97.0 3.0 6

Table 2. ACQUITY UPC 2 mobile phase gradient for MS detection.

Analysis of the Non-Ionic Surfactant Triton-X Using UltraPerformance Convergence Chromatography (UPC2)

3

R E SU LT S A N D D IS C U S S IO N

UV detection results

UPC2 conditions were optimized for the separation and detection of 20 Triton X-100 oligomers. The UV chromatogram for a 10 mg/mL

standard in isopropanol alcohol is shown in Figure 2.

MS detection results

The UV method demonstrated the speed and simplicity of UPC2 for the analysis of Triton X-100. With further optimization of the separation,

in this example using a slower gradient, with MS detection additional characterization of the surfactant was achieved.

The chromatogram for Triton X-100 with MS detection, using the described UPC2 and MS conditions, is shown in Figure 3. The oligomers

detected can be further identified considering the MS spectra, shown in Figure 4 for the oligomers identified as a, b, c, and d in Figure 3.

Figure 2. UV chromatogram for a 10 mg/mL Triton X-100 standard.

Analysis of the Non-Ionic Surfactant Triton-X Using UltraPerformance Convergence Chromatography (UPC2)

4

m/z540 560 580 600 620 640 660 680 700 720 740

%

0

100

m/z540 560 580 600 620 640 660 680 700 720 740

%

0

100

m/z540 560 580 600 620 640 660 680 700 720 740

%

0

100

m/z540 560 580 600 620 640 660 680 700 720 740

%

0

100 709

692

634580559543 552 570 618601590605 620 634 646

657 675674 682693

710

714

715730

716 731 739

665

648

589574543556557 574 591603 612 646630621 634 649

655

666670

671686

672 687 702 732709710 717 733739

621

604

545 546553560 586582571 601605

610

626

627642

628 643 658665 688666 673 689 703704 739724724 733743

577

560

543 552

561

566

578582

583 598584 599 613 659644621622 630 645 660 696679668 681 731697 716722 734743744

b

a

c

d

44

44

44

m/z200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

%

0

100

m/z200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

%

0

100

m/z200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

%

0

100

m/z200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400

%

0

100 709365

214339295

692373

454 634486580

710

714

715

731776 828908 14361017931 1083 1456

343

214 338295

665

648351

454438 589574

666

671

687732 784 877 1036920 1017 13811347 1421 1492

321

214295

621

604329

454440364 545

626

627

658703 820743 997950855 14551106

299

214295

577

560

307

454339 447486

578

583

599659696 1171884774 861 954 1097978 13021206 14611426 1481

Figure 4. Mass spectra for the individual Triton-X oligomers as indicated in Figure 3.

Figure 3. MS chromatogram for a Triton X-100 standard.

Time2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00

%

18

5.725.71

4.98

4.223.51

2.822.81

2.80

2.18

1.67

0.01

1.25

5.74

7.197.21

7.22

7.91

8.62

9.31

9.96

10.56

11.16

11.71

12.26

14.3615.95

16.36

cdb

a

Analysis of the Non-Ionic Surfactant Triton-X Using UltraPerformance Convergence Chromatography (UPC2)

5

By using a slower gradient additional details can be observed, such as the detection of: additional minor series

components, by-products, impurities, degradation products, or contaminants. An additional minor series

present in the analyzed sample of Triton X-100 is shown in Figure 5.

Time

3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50 9.00

%

30

Time0.50 1.50 2.50 3.50 4.50 5.50 6.50 7.50 8.50 9.50 10.50 11.50 12.50 13.50 14.50 15.50 16.50

%

18

5.72

5.714.984.23

4.223.51

3.50

2.822.81

2.80

2.18

1.67

0.01

1.25

1.050.880.22 1.482.08 2.47

2.83

3.19

2.99 3.694.13 4.43

4.74

4.99

5.195.52

5.74

7.196.485.75

5.78

6.185.95

6.21

6.49

6.50

7.17

6.67

6.91

7.21

7.22

7.91

7.417.77

8.62

7.97

8.11

8.42

9.31

8.65

8.67

8.98

9.96

9.34

9.52

10.56

10.16

11.16

10.96

11.71

11.39

12.26

12.08

14.3612.77

12.63 13.14 13.29 15.9514.64 15.5416.36 16.77

g hfe

m/z480 500 520 540 560 580 600 620 640 660

%

0

100

m/z480 500 520 540 560 580 600 620 640 660

%

0

100

m/z480 500 520 540 560 580 600 620 640 660

%

0

100

m/z480 500 520 540 560 580 600 620 640 660

%

0

100 620.58

485.99603.68569.35491.15 542.39

621.58

626.61

576.59

485.82 559.55489.89 547.20

577.58

582.61 613.61 639.05

532.55

485.95515.51

489.85

537.53

538.49 561.07 604.63 660.81646.85

488.51

493.53

509.40 659.64570.91542.47 635.06603.07

f

e

g

h

44

44

44

Figure 5. Additional minor series highlighted in the analyzed sample of Triton X-100, with respective mass spectra.

Analysis of the Non-Ionic Surfactant Triton-X Using UltraPerformance Convergence Chromatography (UPC2)

Waters Corporation 34 Maple Street Milford, MA 01757 U.S.A. T: 1 508 478 2000 F: 1 508 872 1990 www.waters.com

Waters, UPLC, UPC,2 ACQUITY UPC,2 MassLynx, Empower, and The Science of What’s Possible are registered trademarks of Waters Corporation. UltraPerformance Convergence Chromatography is a trademark of Waters Corporation. All other trademarks are the property of their respective owners.

©2015 Waters Corporation. Produced in the U.S.A. September 2015 720005496EN AG-PDF

CO N C LU S IO NS■■ Rapid, high efficiency separation with analysis time of less than 2 min

with PDA detection.

■■ Excellent resolution for approximately 20 oligomers.

■■ Analysis occurs at lower temperature than in GC or SFC.

■■ Reduction in consumption of organic solvents.

■■ MS detection can be used to further characterize the surfactant, such as the

identification of specific oligomers, detection of additional series components,

by-products, impurities, degradation products of contaminants.

References

1. R A Escott, S J Brinkworth, T A Steedman. The determination of ethoxylate oligomers distribution of Non-Ionic and Anionic Surfactants by HPLC. J Chromatography. 282: 655–661, 1983.

2. K Nakamura, Y Morikawa, I Matsumoto. Rapid Analsyis of Ionic and Non-Ionic Surfactants Homologs by HPLC. Journal of the American Oil Chemists’ Society. 58: 72–77, 1981.

3. B J Hoffman, L T Taylor. A Study of Polyethoxylated Alkylphenols by Packed Column Supercritical Fluid Chromatography. Journal of Chromatography. 40: 61–68; Feb 2002.

4. C Bor Fuh, M Lai, H Y Tsai, C M Chang. Impurity analysis of 1,4-dioxane in nonionic surfactants and cosmetics using headspace solid-phase microextraction coupled with gas chromatography and gas chromatography-mass spectrometry. Journal of Chromatography A. 1071: 141–145; 2005.

5. J A Field, D J Miller, T M Field, S B Hawthorne, W Giger. Quantitative determination of sulfonated aliphatic and aromatic surfactants in sewage sludge by ion-pair/supercritical fluid extraction and derivatization gas chromatography/mass spectrometry. Analytical Chemistry. 64(24): 3161–3167; 1992.