STKM1121 Food Chemical Laboratory Sem 1 2022/2023

Name: Adrianna Beathannie Michelle Anak Asin

Matric Number: A187742
Group Number: 7
Title: Peroxidase Assay to Determine Adequacy of Blanching

Objectives:
1. To compare the peroxidase activity of samples based on different temperature.
2. To determine the relationship between peroxidase activity and blanching time.

Introduction:
Blanching is a process in which a food, normally fruits or vegetables are submerged in
boiling water and the water is removed after a period and finally plunged into ice water or
placed under cold running tap water to stop the process. Blanching vegetables before freezing
can stop the enzyme action that may causes loss of colour, flavour and texture when it places in
the freezer. Besides that, blanching also cleanses the surface from dirt and microbes, brightens
the colour and helps to retard loss of vitamins. It softens the vegetables and make them easier
to pack. It is a heat treatment applied depends upon the specificity of the vegetables.
There are two important heat resistance enzymes in the vegetables which are
peroxidase and catalase. If these enzymes are destroyed, the other enzymes in the vegetables
will also be inactivated. Peroxidase is an enzyme widely found in plants, humans and bacteria.
They involve in many physiological processes in plants such as the biosynthesis of lignin in
which make the plant more rigid and cause the cell wall to become hydrophobic and responses
to biotic and abiotic stresses. They are also involved in the reactive oxygen species scavenging
to prevent oxidation from damaging the cell. In this experiment, the function of peroxidase is to
break down hydrogen peroxide (H₂O₂), the toxins produced as a by-product of oxygen during
respiration. The activity of peroxidase can be obtained when the H₂O₂ is break down into water
and alcohol.
Result:

BLANCHING TEMPERATURE: 60˚C

Absorbance Replication
Blanching
Reading
Time (sec) 1 2 3 Mean
(min)

0 0.071 0.074 0.069 0.0713

1 0.07 0.073 0.07 0.0710

2 0.07 0.073 0.071 0.0713

3 0.07 0.073 0.073 0.0720

4 0.07 0.073 0.074 0.0723

BLANK 5 0.07 0.073 0.075 0.0727

6 0.07 0.074 0.076 0.0733

7 0.07 0.074 0.078 0.0740

8 0.07 0.074 0.077 0.0737

9 0.07 0.073 0.078 0.0737

10 0.07 0.074 0.08 0.0747

0 2.562 2.616 2.597 2.5917

1 2.715 2.714 2.787 2.7387

2 2.819 2.878 2.847 2.8480

3 2.991 3.038 2.951 2.9933

4 3.039 3.089 3.039 3.0557

0 5 3.089 3.15 3.149 3.1293

6 3.149 3.147 3.215 3.1703

7 3.148 3.214 3.215 3.1923

8 3.294 3.391 3.295 3.3267

9 3.215 3.392 3.294 3.3003

10 3.295 3.392 3.293 3.3267

 0 2.448 2.501 2.543 2.4973

1 2.53 2.629 2.647 2.6020

2 2.593 2.786 2.785 2.7213

3 2.669 2.816 2.877 2.7873

4 2.692 2.914 2.913 2.8397

50 (R1) 5 2.738 2.993 2.95 2.8937

6 2.668 3.036 2.993 2.8990

7 2.628 3.037 3.088 2.9177

8 2.609 3.089 3.091 2.9297

9 2.578 3.145 3.216 2.9797

10 2.515 3.147 3.146 2.9360

0 1.484 1.474 1.456 1.4713

1 1.557 1.54 1.516 1.5377

2 1.616 1.591 1.553 1.5867

3 1.668 1.643 1.603 1.6380

4 1.693 1.695 1.651 1.6797

50 (R2) 5 1.726 1.723 1.694 1.7143

6 1.753 1.778 1.722 1.7510

7 1.801 1.803 1.761 1.7883

8 1.84 1.83 1.793 1.8210

9 1.833 1.865 1.808 1.8353

10 1.847 1.905 1.819 1.8570

0 1.184 1.157 1.172 1.1710

1 1.289 1.28 1.29 1.2863

100 (R1)
2 1.379 1.345 1.369 1.3643

3 1.46 1.428 1.459 1.4490

 4 1.542 1.51 1.543 1.5317

5 1.615 1.574 1.625 1.6047

6 1.681 1.647 1.703 1.6770

7 1.734 1.693 1.761 1.7293

8 1.799 1.742 1.786 1.7757

9 1.839 1.796 1.84 1.8250

10 1.89 1.833 1.874 1.8657

0 0.726 0.746 0.711 0.7277

1 0.776 0.799 0.761 0.7787

2 0.821 0.846 0.808 0.8250

3 0.87 0.887 0.846 0.8677

4 0.902 0.928 0.886 0.9053

100 (R2) 5 0.946 0.971 0.919 0.9453

6 0.973 1.004 0.951 0.9760

7 1.004 1.03 0.981 1.0050

8 1.034 1.045 0.998 1.0257

9 1.063 1.077 1.022 1.0540

10 1.088 1.102 1.045 1.0783

0 0.884 0.899 0.896 0.8930

1 1.021 1.043 1.029 1.0310

2 1.139 1.184 1.159 1.1607

3 1.274 1.278 1.274 1.2753

150 (R1)
4 1.372 1.387 1.386 1.3817

5 1.5 1.474 1.471 1.4817

6 1.568 1.57 1.583 1.5737

7 1.653 1.642 1.651 1.6487

 8 1.734 1.735 1.749 1.7393

9 1.804 1.815 1.823 1.8140

10 1.863 1.891 1.867 1.8737

0 0.656 0.616 0.662 0.6447

1 0.738 0.697 0.753 0.7293

2 0.804 0.768 0.829 0.8003

3 0.861 0.83 0.893 0.8613

4 0.914 0.886 0.948 0.9160

150 (R2) 5 0.956 0.929 0.998 0.9610

6 0.998 0.971 1.042 1.0037

7 1.034 1.008 1.084 1.0420

8 1.069 1.043 1.119 1.0770

9 1.101 1.074 1.157 1.1107

10 1.133 1.105 1.187 1.1417

0 0.836 0.843 0.869 0.8493

1 0.964 0.97 1.018 0.9840

2 1.079 1.079 1.125 1.0943

3 1.198 1.186 1.229 1.2043

4 1.275 1.284 1.333 1.2973

200 (R1) 5 1.363 1.372 1.409 1.3813

6 1.44 1.457 1.497 1.4647

7 1.567 1.528 1.571 1.5553

8 1.575 1.597 1.632 1.6013

9 1.639 1.65 1.696 1.6617

10 1.689 1.777 1.748 1.7380

200 (R2) 0 0.561 0.683 0.68 0.6413

 1 0.64 0.773 0.764 0.7257

2 0.702 0.854 0.84 0.7987

3 0.761 0.927 0.906 0.8647

4 0.812 0.988 0.971 0.9237

5 0.862 1.043 1.027 0.9773

6 0.908 1.097 1.077 1.0273

7 0.948 1.142 1.124 1.0713

8 0.984 1.197 1.172 1.1177

9 1.021 1.24 1.219 1.1600

10 1.051 1.288 1.253 1.1973

0 0.996 0.953 0.94 0.9630

1 1.036 0.995 0.982 1.0043

2 1.111 1.033 1.018 1.0540

3 1.125 1.059 1.052 1.0787

4 1.141 1.109 1.099 1.1163

250 (R1) 5 1.161 1.13 1.138 1.1430

6 1.183 1.187 1.186 1.1853

7 1.217 1.206 1.221 1.2147

8 1.236 1.204 1.226 1.2220

9 1.27 1.207 1.257 1.2447

10 1.275 1.22 1.234 1.2430

0 2.349 2.368 2.328 2.3483

1 2.459 2.501 2.447 2.4690

250 (R2) 2 2.577 2.575 2.514 2.5553

3 2.632 2.669 2.63 2.6437

4 2.69 2.712 2.692 2.6980

 5 2.735 2.786 2.816 2.7790

6 2.845 2.844 2.818 2.8357

7 2.879 2.951 2.818 2.8827

8 2.911 2.992 2.952 2.9517

9 2.99 2.991 2.992 2.9910

10 2.992 3.038 2.952 2.9940

0 1.33 1.433 1.476 1.4130

1 1.441 1.495 1.554 1.4967

2 1.539 1.545 1.634 1.5727

3 1.612 1.608 1.715 1.6450

4 1.678 1.649 1.827 1.7180

300 (R1) 5 1.74 1.67 1.855 1.7550

6 1.81 1.68 1.912 1.8007

7 1.865 1.693 1.928 1.8287

8 1.902 1.701 1.999 1.8673

9 1.926 1.704 1.979 1.8697

10 1.942 1.696 1.968 1.8687

0 1.184 1.141 1.136 1.1537

1 1.264 1.202 1.193 1.2197

2 1.301 1.251 1.24 1.2640

3 1.342 1.296 1.293 1.3103

300 (R2) 4 1.385 1.343 1.331 1.3530

5 1.423 1.377 1.365 1.3883

6 1.456 1.437 1.393 1.4287

7 1.47 1.44 1.43 1.4467

8 1.504 1.49 1.518 1.5040

 9 1.557 1.512 1.517 1.5287

10 1.579 1.518 1.525 1.5407

BLANCHING TEMPERATURE: 70˚C

Absorbance
Blanching
Reading Replication
Time (sec)
(min)

1 2 3 Mean

BLANK 0 0.063 0.054 0.058 0.0583

1 0.062 0.054 0.06 0.0587

2 0.063 0.054 0.058 0.0583

3 0.062 0.054 0.058 0.0580

4 0.064 0.054 0.059 0.0590

5 0.064 0.056 0.059 0.0597

6 0.063 0.055 0.059 0.0590

7 0.063 0.055 0.059 0.0590

8 0.063 0.055 0.059 0.0590

9 0.065 0.055 0.06 0.0600

10 0.064 0.057 0.059 0.0600

50 (R1) 0 0.63 0.596 0.607 0.6110

1 0.684 0.647 0.686 0.6723

2 0.736 0.688 0.697 0.7070

3 0.776 0.729 0.754 0.7530

4 0.809 0.761 0.783 0.7843

5 0.842 0.786 0.815 0.8143

6 0.863 0.808 0.847 0.8393

 7 0.894 0.815 0.872 0.8603

8 0.917 0.841 0.898 0.8853

9 0.938 0.864 0.911 0.9043

10 0.954 0.884 0.927 0.9217

50 (R2) 0 0.789 0.787 0.791 0.7890

1 0.865 0.851 0.858 0.8580

2 0.926 0.91 0.911 0.9157

3 0.969 0.95 0.96 0.9597

4 1.009 0.988 1.004 1.0003

5 1.038 1.028 1.044 1.0367

6 1.059 1.058 1.08 1.0657

7 1.088 1.086 1.107 1.0937

8 1.106 1.114 1.128 1.1160

9 1.117 1.139 1.143 1.1330

10 1.14 1.162 1.161 1.1543

100 (R1) 0 0.721 0.721 0.68 0.7073

1 0.775 0.779 0.743 0.7657

2 0.812 0.827 0.783 0.8073

3 0.831 0.851 0.819 0.8337

4 0.851 0.864 0.847 0.8540

5 0.866 0.89 0.869 0.8750

6 0.884 0.897 0.877 0.8860

7 0.887 0.902 0.886 0.8917

8 0.893 0.898 0.881 0.8907

9 0.89 0.891 0.877 0.8860

10 0.88 0.882 0.868 0.8767

100 (R2) 0 0.762 0.743 0.746 0.7503

1 0.793 0.772 0.779 0.7813

2 0.83 0.796 0.803 0.8097

3 0.849 0.815 0.827 0.8303

4 0.858 0.844 0.85 0.8507

5 0.875 0.884 0.87 0.8763

6 0.882 0.909 0.886 0.8923

7 0.892 0.894 0.9 0.8953

8 0.911 0.906 0.915 0.9107

9 0.918 0.911 0.918 0.9157

10 0.932 0.974 0.922 0.9427

150 (R1) 0 0.62 0.596 0.627 0.6143

1 0.657 0.632 0.665 0.6513

2 0.69 0.661 0.7 0.6837

3 0.72 0.691 0.731 0.7140

4 0.744 0.716 0.759 0.7397

5 0.767 0.736 0.783 0.7620

6 0.787 0.763 0.8 0.7833

7 0.795 0.785 0.821 0.8003

8 0.807 0.792 0.841 0.8133

9 0.822 0.808 0.862 0.8307

10 0.83 0.813 0.866 0.8363

150 (R2) 0 0.56 0.575 0.549 0.5613

1 0.591 0.607 0.578 0.5920

2 0.619 0.633 0.603 0.6183

3 0.643 0.659 0.628 0.6433

 4 0.668 0.681 0.647 0.6653

5 0.688 0.7 0.663 0.6837

6 0.707 0.721 0.674 0.7007

7 0.723 0.736 0.684 0.7143

8 0.738 0.748 0.691 0.7257

9 0.756 0.759 0.704 0.7397

10 0.764 0.767 0.704 0.7450

200 (R1) 0 0.659 0.696 0.668 0.6743

1 0.703 0.742 0.712 0.7190

2 0.725 0.784 0.754 0.7543

3 0.764 0.825 0.788 0.7923

4 0.802 0.857 0.825 0.8280

5 0.833 0.889 0.858 0.8600

6 0.858 0.918 0.886 0.8873

7 0.888 0.948 0.91 0.9153

8 0.91 0.973 0.933 0.9387

9 0.929 0.996 0.958 0.9610

10 0.943 1.016 0.968 0.9757

200 (R2) 0 0.554 0.571 0.548 0.5577

1 0.592 0.608 0.59 0.5967

2 0.624 0.641 0.622 0.6290

3 0.656 0.669 0.651 0.6587

4 0.687 0.698 0.679 0.6880

5 0.717 0.728 0.704 0.7163

6 0.744 0.746 0.73 0.7400

7 0.767 0.77 0.752 0.7630

 8 0.793 0.778 0.773 0.7813

9 0.914 0.792 0. 789 0.7983

10 0.831 0.796 0.801 0.8093

BLANCHING TEMPERATURE: 80˚C

Absorbance Replication
Blanching
Reading
Time (sec) 1 2 3 Mean
(min)

0 0.362 0.344 0.343 0.3497

1 0.377 0.357 0.358 0.3640

2 0.388 0.368 0.366 0.3740

3 0.399 0.377 0.376 0.3840

4 0.407 0.386 0.383 0.3920

50 (R1) 5 0.416 0.392 0.392 0.4000

6 0.423 0.397 0.4 0.4067

7 0.427 0.398 0.407 0.4107

8 0.43 0.404 0.412 0.4153

9 0.435 0.406 0.418 0.4197

10 0.437 0.41 0.421 0.4227

0 0.303 0.295 0.302 0.3000

1 0.314 0.306 0.313 0.3110

2 0.322 0.314 0.319 0.3183

50 (R2) 3 0.33 0.318 0.326 0.3247

4 0.335 0.323 0.332 0.3300

5 0.34 0.329 0.338 0.3357

6 0.343 0.332 0.341 0.3387

 7 0.345 0.336 0.343 0.3413

8 0.35 0.339 0.344 0.3443

9 0.353 0.341 0.346 0.3467

10 0.356 0.342 0.346 0.3480

0 0.387 0.416 0.396 0.3997

1 0.406 0.439 0.417 0.4207

2 0.421 0.458 0.434 0.4377

3 0.435 0.475 0.45 0.4533

4 0.45 0.488 0.463 0.4670

100 (R1) 5 0.46 0.501 0.475 0.4787

6 0.471 0.512 0.487 0.4900

7 0.482 0.522 0.497 0.5003

8 0.491 0.532 0.508 0.5103

9 0.498 0.539 0.516 0.5177

10 0.505 0.543 0.522 0.5233

0 0.333 0.337 0.34 0.3367

1 0.344 0.35 0.354 0.3493

2 0.354 0.362 0.364 0.3600

3 0.363 0.371 0.373 0.3690

4 0.372 0.38 0.38 0.3773

100 (R2) 5 0.38 0.387 0.389 0.3853

6 0.386 0.393 0.393 0.3907

7 0.392 0.395 0.398 0.3950

8 0.398 0.399 0.402 0.3997

9 0.4 0.401 0.406 0.4023

10 0.402 0.402 0.409 0.4043

 0 0.306 0.293 0.303 0.3007

1 0.322 0.308 0.32 0.3167

2 0.336 0.321 0.333 0.3300

3 0.347 0.333 0.344 0.3413

4 0.359 0.343 0.355 0.3523

150 (R1) 5 0.371 0.351 0.364 0.3620

6 0.379 0.359 0.373 0.3703

7 0.388 0.366 0.381 0.3783

8 0.396 0.374 0.388 0.3860

9 0.402 0.378 0.396 0.3920

10 0.408 0.383 0.4 0.3970

0 0.402 0.318 0.283 0.3343

1 0.425 0.33 0.296 0.3503

2 0.442 0.343 0.308 0.3643

3 0.459 0.355 0.322 0.3787

4 0.475 0.367 0.331 0.3910

150 (R2) 5 0.496 0.379 0.34 0.4050

6 0.498 0.385 0.348 0.4103

7 0.516 0.394 0.354 0.4213

8 0.519 0.399 0.361 0.4263

9 0.526 0.405 0.365 0.4320

10 0.532 0.411 0.371 0.4380

0 0.234 0.231 0.228 0.2310

1 0.271 0.267 0.267 0.2683

200 (R1)
2 0.298 0.294 0.295 0.2957

3 0.323 0.316 0.315 0.3180

 4 0.341 0.336 0.334 0.3370

5 0.357 0.352 0.351 0.3533

6 0.372 0.368 0.366 0.3687

7 0.384 0.379 0.378 0.3803

8 0.399 0.392 0.388 0.3930

9 0.406 0.401 0.4 0.4023

10 0.415 0.411 0.407 0.4110

0 0.315 0.32 0.321 0.3187

1 0.336 0.342 0.342 0.3400

2 0.353 0.358 0.358 0.3563

3 0.367 0.373 0.373 0.3710

4 0.38 0.387 0.387 0.3847

200 (R2) 5 0.391 0.397 0.399 0.3957

6 0.403 0.409 0.409 0.4070

7 0.413 0.417 0.419 0.4163

8 0.421 0.426 0.427 0.4247

9 0.428 0.434 0.434 0.4320

10 0.434 0.441 0.439 0.4380

0 0.343 0.331 0.34 0.3380

1 0.371 0.358 0.366 0.3650

2 0.395 0.38 0.389 0.3880

3 0.416 0.4 0.41 0.4087

250 (R1)
4 0.436 0.419 0.428 0.4277

5 0.454 0.437 0.446 0.4457

6 0.468 0.451 0.459 0.4593

7 0.484 0.466 0.474 0.4747

 8 0.499 0.479 0.488 0.4887

9 0.511 0.491 0.5 0.5007

10 0.52 0.504 0.51 0.5113

0 0.419 0.372 0.389 0.3933

1 0.456 0.404 0.422 0.4273

2 0.488 0.432 0.451 0.4570

3 0.516 0.457 0.478 0.4837

4 0.542 0.479 0.5 0.5070

250 (R2) 5 0.567 0.5 0.522 0.5297

6 0.589 0.519 0.542 0.5500

7 0.607 0.538 0.562 0.5690

8 0.625 0.554 0.58 0.5863

9 0.644 0.567 0.595 0.6020

10 0.662 0.577 0.608 0.6157

0 0.397 0.388 0.397 0.3940

1 0.432 0.424 0.434 0.4300

2 0.465 0.457 0.466 0.4627

3 0.493 0.486 0.494 0.4910

4 0.518 0.514 0.522 0.5180

300 (R1) 5 0.542 0.536 0.545 0.5410

6 0.558 0.557 0.57 0.5617

7 0.575 0.576 0.593 0.5813

8 0.591 0.597 0.613 0.6003

9 0.604 0.617 0.631 0.6173

10 0.621 0.633 0.648 0.6340

300 (R2) 0 0.477 0.482 0.475 0.4780

 1 0.507 0.513 0.507 0.5090

2 0.537 0.539 0.535 0.5370

3 0.56 0.564 0.561 0.5617

4 0.582 0.585 0.584 0.5837

5 0.601 0.606 0.605 0.6040

6 0.614 0.624 0.622 0.6200

7 0.618 0.638 0.639 0.6317

8 0.631 0.653 0.657 0.6470

9 0.646 0.666 0.67 0.6607

10 0.655 0.676 0.683 0.6713

Graph of absorbance vs kinetic rate (minutes)

For blanching at 60°C,

Absorbance vs Kinetic Rate

4
Absorbance

3
2
1
0
0 2 4 6 8 10 12

y = 0.0713x + 2.6389 y = 0.0399x + 1.9928 Kinetic Rate (Minutes)

R² = 0.9279 R² = 0.9143 y = 0.0514x + 0.9256 y = 0.0725x + 0.761 y = 0.0704x + 0.734
y = 0.0467x + 1.6379 y = 0.0422x + 1.2674 R² = 0.9814 R² = 0.9837 R² = 0.9851
R² = 0.9692 R² = 0.9568

0 50 100 150
200 250 300 Linear (0)
Linear (0) Linear (50) Linear (50) Linear (100)
Linear (100) Linear (150) Linear (150) Linear (200)
Linear (200) Linear (250) Linear (250) Linear (300)
For blanching at 70°C,

Absorbance vs Kinetic Rate

1.2
y = 0.0323x + 0.6756 50
R² = 0.9612
1 100
y = 0.0166x + 0.7027
R² = 0.8757 150
0.8 y = 0.0199x + 0.5444
Absorbance

200
R² = 0.9654
0.6 250
y = 0.0275x + 0.5712
R² = 0.9826 300
0.4
y = 0.0153x + 0.3726 Linear (50)
R² = 0.9914
0.2 Linear (50)
y = 0.0327x + 0.6628
Linear (100)
0 R² = 0.9868
0 2 4 6 8 10 12 Linear (100)
Kinetic Rate (Minutes) Linear (150)

For blanching at 80°C,

Absorbance vs Kinetic Rate

0.7
y = 0.0056x + 0.2689
50
R² = 0.9419
0.6 100
y = 0.0092x + 0.3141
R² = 0.9633 150
0.5
y = 0.0096x + 0.2622 200
Absorbance

0.4 R² = 0.9707
y = 0.0141x + 0.2291 250
R² = 0.9603 300
0.3
y = 0.0192x + 0.3166 Linear (50)
R² = 0.9827
0.2
y = 0.0209x + 0.3901 Linear (50)
R² = 0.9782 Linear (100)
0.1
Linear (100)
0
Linear (100)
0 2 4 6 8 10 12
Kinetic Rate (Minutes) Linear (150)
Calculation:
For blanching temperature at 60°C

Blanching duration ΔA (changes in Activity of

(seconds) absorbance/min) peroxidase (U/mL)
0 0.0713 80.41
50 0.0399 45
100 0.0514 57.97
150 0.0725 81.77
200 0.0704 79.40
250 0.0467 52.67
300 0.0422 47.59

For blanching temperature at 70°C

Blanching duration ΔA (changes in Activity of

(seconds) absorbance/min) peroxidase (U/mL)

50 0.0323 36.43

100 0.0166 18.71

150 0.0199 22.44

200 0.0275 31.02

250 0.0153 17.26

300 0.0327 36.88

For blanching temperature at 80°C

Blanching duration ΔA (changes in Activity of

(seconds) absorbance/min) peroxidase (U/mL)

50 0.0056 6.32

100 0.0092 10.38

150 0.0096 10.83

200 0.0141 15.90

250 0.0192 21.65

 300 0.0209 23.57

Peroxidase vs Blanching time

90
80
y = -0.0441x + 70.154
70 60℃
R² = 0.0841
Peroxidase (U/mL)

60
70℃
50
80℃
40
y = 0.0037x + 26.48 Linear (60℃)
30
R² = 0.0015 Linear (60℃)
20
10 y = 0.0715x + 2.262 Linear (70℃)
R² = 0.9631
0 Linear (70℃)
0 50 100 150 200 250 300 350
Linear (80℃)
Blanching duration (sec)
Linear (80℃)

Discussion:
From the graph of activity against blanching time, the long beans blanched at 60°C have
the highest average peroxidase activity followed by those blanched at 70°C and 80°C. The
peroxidase activity is decreased with increasing blanching time. It is an inverse relationship.

Peroxidase is an enzyme which can be found in most of the plant-based food. It may
cause oxidation in organic compounds and lead to deterioration in colour, flavour and
nutritional quality of fruits and vegetables. Blanching is a heat treatment used to inactive the
enzyme. Water blanching and steam blanching are the two common blanching methods used.
Time of blanching is important and varies with the vegetable. Underblanching stimulates the
enzyme activity and is worser than no blanching while overblanching causes loss of colour,
flavours, vitamins and minerals.
Peroxidase is widely used as an indicator of blanch efficacy due to its high
thermostability. The peroxidase enzyme is destroyed when there is heat supply and a lower
peroxidase activity can be obtained. The adequacy blanching in different temperature and
duration were determined based on the peroxidase activity in the long beans. It is a simple test
by using guaiacol (colourless) to tetraguaiacol (yellow brown). The samples with high
peroxidase activity indicate that there are still many peroxidase enzymes present in the long
beans and the blanching temperature or duration is not enough to inhibit the peroxidase
enzyme.
The long beans blanched at 80°C and in duration 300 seconds have the lowest
peroxidase activity. This is due to a higher temperature and longer blanching period can
inactive or even denature the peroxidase enzyme. Hence, the peroxidase activity will be lower.
This show that the long beans blanched in a higher temperature and longer duration is effective
than those at lower temperature and shorter blanching duration.
Some precautions should be taken to obtain a more accurate result. First, the water
bath must be maintained at the desired temperature to ensure the blanching temperature is
constant along the blanching period. Next, after adding the guaiacol, we should stopper the
cuvette immediately and inverted it 1 or 2 times. The experiment needs to repeat at least twice
to get the average absorbance for a more accurate result. Last, the concentration of peroxidase
may not distribute consistently for each long beans and this may lead to inaccuracy of result.
Conclusion:
The long beans blanched at 60°C has the highest average of peroxidase activity compared to
those blanched at 70°C and 80°C. The peroxidase activity and blanching time have an inverse
relationship which the peroxidase activity decreases with increasing in blanching time. The
most adequacy blanching temperature is 80°C and duration is 300 seconds to inhibit the activity
of peroxidase.
Question:
1. What are the enzymes responsible for the deterioration in palatability, colour and
ascorbic acid content?
Lipoxygenase is the enzyme that is responsible for deterioration in palatability in fruits
and vegetables. It is an enzyme that is found in many plants, which catalyzes the
oxygenation of polyunsaturated fatty acids (PUFA) to form fatty acid hydroperoxides
which decompose and form secondary oxidation products that give strong undesirable
flavours, leading to food deterioration. Moreover, the polyphenol oxidase (PPO) that
involve in enzymatic browning reaction may affect the colour in the fruits and
vegetables. PPO converts phenolic compounds through oxidation into quinones. After
that it react with other compounds to form melanin and form brown pigment. The
chlorophyllase (a thylakoid membrane glycoprotein) can be found in green plants. This
can also cause the discolouration of green colour in some vegetables and destruction of
chlorophyll. Ascorbic acid oxidase which is a copper-containing enzyme will catalyze the
oxidation of ascorbic acid to yield dehydroascorbic acid and causing loss of Vitamin C in
the fruits and vegetables.

2. Besides blanching, what are other means to reduce quality deterioration in fruits and
vegetables?
 Besides blanching, freezing, fresh and cold storage, freeze drying, canning, drying,
irradiation, pasteurization and preservation with chemical substances such as sugar and
acid are the other means to reduce quality deterioration in fruits and vegetables. Each
of them has their own benefits and disadvantages. Freeze drying is considered to be the
best mean to reduce the quality deterioration in fruits and vegetables since it can
maintain most of the texture, colour, flavour and nutritional value.

Reference:
1. Mamot Said, Abdul Salam Babji, Liew Siew Ling, 2017, Chemical Analysis of Food,
Penerbit Universiti Kebangsaan Malaysia Bangi, page 117-118.
2. Anonymous, Determination of blanch efficacy by means of a rapid colorimetric
peroxidase assay, Biochemical Education 1985 13(2), page 85-86.

3. Michael Ruhlman (January 20, 2009). "Blanching, shocking, refreshing". ruhlman.com.

Retrieved 2018-05-19.