analysis of biocides using uplc · mit cmit carbendazim bit phenoxyethanol benzoic acid...

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Meet Regulatory Mandates to Protect

Methylparaben

Benzoic acid

MIT

CMIT

Carbendazim

BIT

Phenoxyethanol

ANALYSIS OF BIOCIDES USING UPLC®

Peter J. Lee and Alice J. Di Gioia Waters Corporation, Milford, MA, USA

What Do They Have in Common? Solution for Biocide Analysis

S

N

O

S

N

O

Cl

SNH O

NH

N

NH

O

O

O

OH

O OH

OH

O O

1a 1b2

3 4 5 6

MITCMIT

Carbendazim

BIT Phenoxyethanol Benzoic acid Methylparaben

Kathon™

MIT C

MIT

Car

bend

azim

BIT

Phe

noxy

etha

nol B

enzo

ic a

cid

Met

hylp

arab

en

AU

0.00

0.05

0.10

Minutes0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00

Kathon

5 ppm

1

2

3

4

5

6

7

8

9

Mean

Std. Dev .

% RSD

MIT(min)

CMIT(min)

Carbendazim(min)

BIT(min)

Phenoxyethanol(min)

Benzoic acid(min)

Methy lparaben(min)

0.286

0.286

0.287

0.286

0.286

0.286

0.287

0.286

0.287

0.286

0.001

0.19

0.908

0.908

0.908

0.908

0.908

0.908

0.908

0.908

0.909

0.908

0.000

0.04

1.147

1.147

1.147

1.148

1.148

1.149

1.148

1.150

1.150

1.148

0.001

0.10

1.600

1.601

1.601

1.602

1.601

1.602

1.601

1.602

1.602

1.601

0.001

0.03

2.141

2.141

2.141

2.142

2.141

2.142

2.141

2.142

2.142

2.142

0.001

0.03

2.400

2.401

2.400

2.401

2.400

2.401

2.400

2.401

2.401

2.401

0.001

0.02

2.671

2.672

2.671

2.672

2.671

2.672

2.671

2.672

2.672

2.672

0.001

0.02

Table 1.Component Summary For Retention Time

1

2

3

4

5

6

7

8

9

Mean

Std. Dev .

% RSD

MIT CMIT Carbendazim BIT Phenoxyethanol Benzoic acid Methy lparaben

34745

34730

34684

34741

34761

34673

34753

34781

34782

34739

38

0.1

68748

68894

69423

69168

68952

69132

68903

68736

69050

69001

219

0.3

134846

134698

134511

135187

134533

134817

135014

135018

134694

134813

229

0.2

227719

228692

227840

228238

228331

228461

228616

227710

228489

228233

383

0.2

57857

57882

57682

57388

58008

57802

57863

57845

57809

57793

174

0.3

172510

173440

170783

173125

170433

170579

172557

170954

172072

171828

1157

0.7

173458

172053

172192

172113

172156

171725

171723

171833

172143

172155

523

0.3

Table 2.Component Summary For Area

1

2

3

4

5

6

7

Component Match1Spect. Name

Match1Angle

Match1Threshold

Purity1Angle

Purity1Threshold

MIT

CMIT

Carbendazim

BIT

Phenoxyethanol

Benzoic acid

Methy lparaben

2-methy l-4-isothiazolin-3-one

5-chloro-2-methy l-4-isothiazoline-3-one

carbendazim

1,2-benzisothiazol-3-one

2-phenoxyethanol

benzoic acid

methy lparaben

0.488

0.293

0.169

0.181

0.820

0.217

0.288

1.585

1.465

1.239

1.277

2.663

1.387

1.385

0.705

0.694

0.320

0.371

2.062

0.508

0.465

1.035

0.842

0.546

0.616

2.449

0.703

0.726

PDA library matching results table

< <

Calibration Plot

Name: Kathon; R 2̂: 0.999934Name: Carbendazim; R 2̂: 0.999941Name: BIT; R 2̂: 0.999919Name: Phenoxyethanol; R 2̂: 0.999958Name: Benzoic acid; R 2̂: 0.999800Name: Methylparaben; R 2̂: 0.999979

Area

0

100000

200000

300000

400000

500000

600000

700000

800000

900000

Amount0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00

(ppm)

BIT

Methylparaben

Benzoic acid

Carbendazim

Kathon

Phenoxyethanol

ACQUITY UPLC® System rapidly and accurately analyzes biocides

● Separate 7 components in 3 minutes ♦ Baseline resolution

● Excellent reproducibility ● Great sensitivity

Empower™ data automation, from collection to reporting

● Peak purity & identification ● Calibration & quantification ● Custom calculations ● Automate report generation

Ideal solution for: QC, product development, and problem-solving

Experiment:

Instrument and Operation Conditions

LC: ACQUITY UPLC® with PDA Software: Empower™ Column: ACQUITY UPLC BEH C18 2.1 mm x 50 mm Weak Wash: 95:5 H2O/CH3CN (600 µL) Strong Wash: 50:50 H2O/CH3CN (600 µL) Mobile Phase A: 0.05v% of TFA in H2O Mobile Phase B: 0.05v% of TFA in CH3CN Linear Gradient: 5% B to 15% B in 2.9 min Flow Rate: 1 mL/min Sampling Rate: 20 pt/s Injection Volume: 5 µL Column Temperature: 30 °C PDA: 210-500 nm PDA Filter Response: 0.1 sec

Overlay 9 Replicate Injections: PDA Timed Chromatograms

UPLC® RT Reproducibility

UPLC® Area Reproducibility

1. P. J. Lee, A. J. Di Gioia, ACQUITY UPLC PDA ANALYSIS OF BIOCIDES (PART 1), Waters Corporation, Application Note 720001832EN, 2006.

2. P. J. Lee, A. J. Di Gioia, ACQUITY UPLC PDA ANALYSIS OF BIOCIDES (PART 2): PASS OR FAIL CUSTOM CALCULATION, Waters Corporation, Application Note 720001833EN, 2006.

Available for download at www.waters.com

References

Conclusions

UV/PDA Calibration Curves

3-minute analysis of a blend of common biocides

Biocides Kill Micro-organisms

UV Spectra

Extracted from PDA Chromatograms for Library & Peak Purity: ● Confirm peak identity ● Assure non-coelution of spectrally dissimilar components

Protected by Biocides!

No biocides With biocides

Needs for Biocide Analysis

Sensitivity

Resolution

Speed

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