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Effect of oxidation and high pressure treatment on fruit and vegetable juices deallergization


Key words Apple, carrot, celery, allergens, high pressure processing
Latest version 2012/11/14
Completed by FRIP

How does it work?

Primary objective While the primary objective of high pressure treatment is the pasteurization of packed food products at low to ambient temperatures, in some specific cases (e.g. apple juice), the technology has been shown to reduce the allergenicity of fruit and vegetable products. But this idea was not confirmed by further research. The detailed studies did not confirm the effect of high pressure on immunoreactivity and allergenicity of main allergens and juices from original fruit and vegetables (apple, carrot and celery). During studies it was found that some oxidative process assisted with high pressure can be successfully applied.
Working principle Thesis [1] presented interesting results on the effect of high pressure treatment on allergenicity and structural changes of apple, main apple allergen Mal d 1, hazelnut, carrot, cherry, peach and celery. These promising results motivated other researchers to study the effect of high pressure treatment on allergens because if this technology is successful it opens new opportunities for novel technology to be applied for.

But other studies based on a battery of tests made on pure allergens and natural fruit and vegetable juices did not confirm the results of the thesis mentioned above [2-7].

  • Paper [2] dealt with buffered solutions of recombinant Bet v 1 and birch pollen extract.
  • Paper [3] dealt with buffered recombinant Api g 1 solution (main celery allergen).
  • Paper [4] dealt with recombinant main carrot allergen rDau c 1 and carrot juice that were treated by 500 MPa for 10 minutes and different temperatures (30°C, 40°C, 50°C) and also pressures 400 to 550 MPa for 3 and 10 minutes.
  • Paper [5] dealt with extract of Golden Delicious apple treated at various pressures up 800 MPa and stored for 10 months.
  • Paper [6] dealt with apple and carrot main allergens rMal d 1 and rDau c 1. Substances were treated by 400 - 550 MPa held for 3 and 10 minutes at 25°C and at 500 MPa for 10 minutes at 30 - 50°C.
  • Paper [7] treated main apple allergen rMal d 1 by high pressure at 500 MPa for 10 minutes at 30°C and showed the great changes in CD spectra when compared with the untreated samples that evidenced in structural changes. This is in agreement with conclusion of paper [9]. But these publications did not confirmed the de-allergization of products simply by high pressure treatment.

Mechanism Possible mechanism of successful de-allergization of apple slices by high pressure referred to in [1] can be speeding up the chemical reactions connected with apple components oxidative processes. Present phenolic acids are oxidized by present oxygen by activity of polyphenoloxidase [11]. Oxidation products can polymerize with allergenic proteins to form larger structures that hide the epitopes and minimize the allergenic reaction. This process was not observed in celery and carrot juices because some components of above mechanisms are missing or strong antioxidants are present (case of carrot where present carrotenoids stabilize allergenic proteins). This mechanism was successfully applied in apple juice. The juice was oxidized first by mixing with air for one hour and then vitaminized by ascorbic acid to enhance back the nutritional quality destroyed partly by oxidation. The resulting product can be pasteurised regardless method used (heat treatment or high pressure). HP pasteurisation is better because it saves vitamins, colour and other quality parameters.

Combining high pressure and relatively high temperatures (high pressure assisted sterilization process) can substantially lower the immunoreactivity on apple and celeriac [10]. But consider, for apple juices, the high pressure is not necessary tool for deallergization of the substance. The main tool is the oxidation. High pressure treatment is only the method of juice pasteurisation.

Additional effects Microbial stabilization
Important process parameters High presssure in the range of 400-600 MPa. Mixing with air for oxidation held for one hour as minimum.
Important product parameters Presence of phenolic acids, pH, and presence of polyphenoloxidase, oxygen, and low content of antioxidants are the most important parameters for successful deallergization by oxidation. Then the pasteurizastion is received by high pressure treatment.

What can it be used for?

Products Only apple slices and apple juices, or carrot juice mixed with apple juice (ratio 1:5) are known successful cases for high pressure assisted deallergization (internal FRIP report, unpublished).
Operations High pressure pasteurization, high pressure assisted sterilization.
Solutions for short comings The only opportunity to de-allergize apple juices is the oxidation. The high pressure treatment can be used only after deallergization for microbial stabilization (pasteurization).

What can it NOT be used for?

Products Apple juice, apple slices, ev. carrot-apple juices mixture. But, high pressure is used only as pasteurization method, deallergization is done by oxidative process preceding the high pressure treatment.
Operations Not applicable.
Other limitations See important parameters.
Risks or hazards Each batch of deallergized e.g. apple juice should be checked by some quick allergenicity test, e.g. by. ELISA. De-polymerization of complexes and allergenicity recovery was tested during three weeks of deallergized apple juice storage [8]. No recovery was observed.


Maturity In case of apple juice, pilot tests in industrial conditions have been done yet and these products were tested on 19 sensitized patients without any symptomatic reactions. The technology is ready to be applied in industry to produce high quality de-allergized apple juices.
Modularity /Implementation The first step (oxidation process) can be simply implemented in the current production lines (vessel with mixer and sufficient holding time).High pressure treatment needs special equipment.
Consumer aspects As far as tested in apple juice, the overall sensory acceptance by sensitized patients can be characterized by wording “excellent”, “superior”. We can consider that these people are very limited in such juices consumption.
Legal aspects Allergens presence labelling is under the EU legislation regulation.

The EFSA Journal (2004) 32, 1 – 197; Opinion of the Scientific Panel on Dietetic Products, Nutrition and Allergies on a request from the Commission relating to the evaluation of allergenic foods for labelling purposes Directive 2003/89/EC of the European Parliament and of the Council of 10 November 2003 Directive 2000/13/EC of the European Parliament and of the Council of 20 March 2000 Directive 2006/142/EC of the European Parliament and of the Council of 22 December 2006 Directive 2005/26/EC of the European Parliament and of the Council of 21 March 2005

Environmental aspects No substantial impact of this technology on energy consumption or environment is known.

Further Information

Institutes FRIP, IFR, TU München
Companies APA Processing
References [1] Scheibenzuber M. (2003). Molekulare and klinische Auswirkungen einer Hochdruckbehandlung allergener Lebensmittel. PhD thesis, TU Muenchen.

[2] Setinova, I., Kminkova, M., Strohalm, J., Heroldova, M., Novotna, P., Honzova, S., Vavrova, H., Kucera, P., Proskova, A., Houska, M. (2009). Allergenicity of main birch allergen rBet v1 and high-pressure treatment, High Pressure Research, 29, 680-685.

[3] Houska, M., Kminkova, M., Strohalm, J., Setinova, I., Heroldova, M., Novotna, P., Honzova, S., Vavrova, H., Kucera, P., Proskova, A. (2009). Allergenicity of main celery allergen rApi g1 and high-pressure treatment, High Pressure Research, 29, 686-694.

[4] Heroldova, M., Houska, M., Vavrova, H., Kucera, P., Setinova, I., Honzova, S., Kminkova, M., Strohalm, J., Novotna, P., Proskova, A. (2009). Influence of high-pressure treatment on allergenicity of rDau c1 and carrot juice demonstrated by in vitro and in vivo tests, High Pressure Research, 29, 695-704.

[5] Fernandez, A., Butz, P., Tauscher, B. (2009). IgE binding capacity of apple allergens preserved after high pressure treatment, High Pressure Research, 29, 705-712.

[6] Šetinová, I., Honzová, S., Kváčová, A., Trnková, B., Heroldová, M., Vávrová, H., Kučera, P., Houška, M., Kmínková, M., Gabrovská, D., Strohalm, J., Paulíčková, I., Julínek, O., Urbanová, M., (2009). Influence of the high pressure on reduction of allergenicity of proteins rMal d1, rDau c1 and orientation quantification of their contents in apple Golden Delicious and in carrot, Alergie, 11, 102-114 (in Czech)

[7] Houska, M., Heroldova, M., Vavrova, H., Kucera, P., Setinova, I., Havranova, M., Honzova, S., Strohalm, J., Kminkova, M., Proskova, A., Novotna, P. (2009). Is high-pressure treatment able to modify the allergenicity of the main apple juice allergen, Mal d1?, High Pressure Research, 29, 14-22.
[8] Setinova I., Kminkova M., Louckova K., Heroldova M. , Vavrova H., Pruchova J., Strohalm J., Novotna P., Gresova P., Trnkova B., Kvacova A. , Honzova S., Kucera P., Houska M. (2010). Influence of storage and antioxidant on oxidative and polymerisation processes of apple juice and protein Mal d 1, 29th Congress of the European Academy of Allergy and Clinical Imunology, 5 – 9 June 2010, London

[9] Johnson, P.E., Van der Plancken, I., Balasa, A., Husband, F.A., Grauwet, T., Hendrickx, M., Knorr, D., Mills, E.N.C., Mackie, A.R. (2010). High pressure, thermal and pulsed electric-field-induced structural changes in selected food allergens. Molecular Nutrition and Food Research, 54, 1701-1710.

[10] Husband, F.A., Aldick, T., Van der Plancken, I., Grauwet, T., Hendrickx, M., Skypala, I. and Mackie, A.R. (2011) High-pressure treatment reduces the immunoreactivity of the major allergens in apple and celeriac. Molecular Nutrition and Food Research, 55 (7), 1087-1095.

[11] Garcia, A., Wichers , J.H., Wichers H. J. (2007). Decrease of the IgE-binding by Mal d 1, the major apple allergen, by means of polyphenol oxidase and peroxidase treatments, Food Chemistry, 103, 94-100

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Created by M.Houska on 3 February 2012, at 13:20