EP0235528A1 - Process for preparing stable ozonized oils from unsaturated vegetable oils - Google Patents

Process for preparing stable ozonized oils from unsaturated vegetable oils Download PDF

Info

Publication number
EP0235528A1
EP0235528A1 EP87100725A EP87100725A EP0235528A1 EP 0235528 A1 EP0235528 A1 EP 0235528A1 EP 87100725 A EP87100725 A EP 87100725A EP 87100725 A EP87100725 A EP 87100725A EP 0235528 A1 EP0235528 A1 EP 0235528A1
Authority
EP
European Patent Office
Prior art keywords
oil
oils
unsaturated vegetable
ascorbic acid
vegetable oils
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
EP87100725A
Other languages
German (de)
French (fr)
Other versions
EP0235528B1 (en
Inventor
Josef Prof. Dr. Dr. Dipl.-Ing. Washüttl
Renate Dr. Rer. Nat. Dipl.-Chem. Viebahn
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hansler Dr J GmbH
Original Assignee
Hansler Dr J GmbH
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hansler Dr J GmbH filed Critical Hansler Dr J GmbH
Priority to AT87100725T priority Critical patent/ATE42965T1/en
Publication of EP0235528A1 publication Critical patent/EP0235528A1/en
Application granted granted Critical
Publication of EP0235528B1 publication Critical patent/EP0235528B1/en
Expired legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11C—FATTY ACIDS FROM FATS, OILS OR WAXES; CANDLES; FATS, OILS OR FATTY ACIDS BY CHEMICAL MODIFICATION OF FATS, OILS, OR FATTY ACIDS OBTAINED THEREFROM
    • C11C3/00—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom
    • C11C3/006—Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom by oxidation

Definitions

  • the invention relates to a process for the production of stable ozonized oils from unsaturated vegetable oils with removal of physiologically questionable low molecular weight aldehydes and ketones.
  • the process according to the invention provides valuable products which are suitable for therapeutic and cosmetic use in humans and animals.
  • reaction of ozone with unsaturated fatty acids produces peroxidic products as therapeutically valuable active substances and low-molecular aldehydes and ketones, especially malondialdehyde, as undesirable by-products.
  • the peroxidic products are used, for example, in dermatology for fungal diseases, leg ulcers, poorly healing wounds, infected wounds and the like.
  • the ketones and aldehydes formed as by-products are therapeutically ineffective and largely physiologically questionable. The latter is particularly true for malondialdehyde, which occurs in considerable quantities when ozonization of unsaturated vegetable oils is complete (see Registry of Toxic Effects of Chemical Substances, U.S. Department of Health and Human Services, 1984).
  • the object of the invention is to provide ozonized products from unsaturated vegetable oils which are therapeutically valuable products, are stable for a long time and have the lowest possible content of low molecular weight aldehydes and ketones, in particular malondialdehyde.
  • the invention relates to a process for producing stable ozonized oils from unsaturated vegetable oils by introducing an ozone-oxygen mixture into the oil until saturation, which is characterized in that after the ozonization, an extraction treatment in an acidic environment in the presence of a redox system, preferably a biological redox system which is capable of a moderate radical reaction and which carries out in the presence of an artificial or natural antioxidant or reducing agent.
  • a redox system preferably a biological redox system which is capable of a moderate radical reaction and which carries out in the presence of an artificial or natural antioxidant or reducing agent.
  • unsaturated vegetable oils are olive oil, safflower oil, wheat germ oil, linseed oil, almond oil, nut oil, sunflower oil, poppy seed oil, sesame oil, castor oil, croton oil, soybean oil and palm oil. Olive and safflower oils are preferred. Olive oil is especially preferred.
  • the redox system used which is capable of radical reactions, acts in the organism both as an electron acceptor and as an electron donor.
  • Preferred examples of redox systems are ascorbic acid, the essentially membrane-associated redox system vitamin E, vitamin A and the biological quinoid / benzoid systems. Ascorbic acid is particularly preferred.
  • a pH of 3.5 to 6.5 has proven to be particularly suitable for carrying out the extraction treatment.
  • ascorbic acid or citric acid can be used.
  • antioxidants or reducing agents are butylated hydroxyanisole, gallate and hydrogen sulfite.
  • the ozone-oxygen mixture is produced in compliance with the regulations for the production or filling of medical oxygen.
  • the ozone is generated by a silent electrical discharge (absolutely nitrogen-free to avoid the formation of aggressive and reactive nitrogen oxides, especially radicals).
  • the ozonization of the oil is carried out with a continuous pass with a gas flow in the range of 1 to 2 liters per minute.
  • the ozone concentration used is preferably in the range from 50 to 70 ⁇ g per ml.
  • the oil is bubbled through using a cylindrical vessel with the highest possible “water column” with thermostatting at about 20 ° C.
  • a continuous flow is important, especially just before the saturation limit, which becomes apparent when the product solidifies at 20 ° C.
  • For 12 liters with a total height of the cylindrical vessel of 60 cm and a diameter of approx. 20 cm, a uniform bubbling time of 180 to 300 hours is required, depending on the ozone concentration of the initial ozone-oxygen mixture.
  • aqueous extraction solution For the extraction treatment, about 2 to 20, in particular 3 to 10 and preferably about 5 ml of aqueous extraction solution are used per 1 g of ozonized oil.
  • the extraction is preferably carried out by shaking for 10 to 60 minutes. The shaking process can be carried out several times to increase the effectiveness. Any excess sulfite still present in the extract can be removed by shaking out the oily phase with 2% aqueous ascorbic acid solution.
  • Particularly preferred combinations for the extraction treatment according to the invention are ascorbic acid + butylated hydroxyanisole, citric acid and butylated hydroxyanisole as well as ascorbic acid and hydrogen sulfite.
  • the determination is based on the reaction of malondialdehyde with 2-methylindole in alcoholic-hydrochloric acid solution to form a stable, intensely colored complex.
  • Determination of malondialdehyde in ozonized oil About 1 g of the solid oil is weighed exactly and warmed to liquefy under an N2 atmosphere (approx. 30 ° C). Since the oil is immiscible with the reagents, 1 ml of distilled H2O are added in addition to 2 ml of methyl indole and stirred with a magnetic stirrer under the N2 atmosphere during the 20 min reaction time to allow a complete reaction of the malondialdehyde. The alcoholic phase is then measured (after the fat has been removed) in comparison with the calibration series that is applied in parallel each time.
  • the product obtained has a peroxide number (POZ) of 929 and a malondialdehyde content of 445 ⁇ g / g of oil.
  • the extraction treatment removed 85 percent of the malondialdehyde, while the peroxide number only decreased by 5.7 percent.
  • the aqueous extract contains no peroxides, while the malondialdehyde has almost completely passed into the aqueous phase.
  • Samples of the ozonated olive oil produced according to Example 1 are subjected to different extraction treatments. For this purpose, 1 g ozonized oil is shaken out with various aqueous solutions for 15 minutes. The POZ and malondialdehyde content are then determined. The results are summarized in the table below.
  • x) 1 means that the parameter in question has not been determined
  • x) 2 excess sulfite was completely removed by shaking the oil with 10 ml H2O (2 g ascorbic acid / 100 ml), whereby the POZ and malondialdehyde content of the ozonated oil did not change significantly: the peroxide number decreased from 876 to 860, the malondialdehyde content from 68 ⁇ g / g to 52 ⁇ g / g.

Landscapes

  • Chemical & Material Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Fats And Perfumes (AREA)
  • Cosmetics (AREA)
  • Medicinal Preparation (AREA)

Abstract

1. Method of producing stable ozonised oils from unsaturated vegetable oils by introducing an ozone-oxygen mixture into the oil to saturation, characterized in that, after the ozonising, there is effected an extraction treatment in acid environment in the presence of a redox system, which is capable of a radical reaction, and in the presence of a synthetic or natural anti-oxidant or reducing agent.

Description

Die Erfindung betrifft ein Verfharen zur Her­stellung von stabilen ozonisierten Ölen aus ungesät­tigten Pflanzenölen unter Entfernung von physiolo­gisch bedenklichen niedermolekularen Aldehyden und Ketonen. Das erfindungsgemäße Verfahren liefert wert­volle Produkte, die sich zur therapeutischen und kos­metischen Anwendung bei Mensch und Tier eignen.The invention relates to a process for the production of stable ozonized oils from unsaturated vegetable oils with removal of physiologically questionable low molecular weight aldehydes and ketones. The process according to the invention provides valuable products which are suitable for therapeutic and cosmetic use in humans and animals.

Die keimtötende Wirkung von ozonisiertem Oli­venöl ist seit längerer Zeit bekannt (vgl. beispiels­weise G. Cronheim, Journal of the American Pharma­ceutical Association, Bd. 36 (1947), S. 274). Ent­sprechende Handelsprodukte wurden jedoch wegen ihrer leichten Zersetzbarkeit bald wieder aus dem Handel gezogen (vgl. M. Schönbauer, OzoNachrichten, Bd. 3 (1984), S. 28).The germicidal effect of ozonized olive oil has been known for a long time (see, for example, G. Cronheim, Journal of the American Pharmaceutical Association, Vol. 36 (1947), p. 274). Corresponding commercial products, however, were soon withdrawn from the market because of their easy decomposability (cf. M. Schönbauer, OzoNachrichten, Vol. 3 (1984), p. 28).

Bei der Umsetzung von Ozon mit ungesättigten Fettsäuren entstehen als therapeutisch wertvolle Wirkstoffe peroxidische Produkte und als unerwünsch­te Nebenprodukte niedermolekulare Aldehyde und Ke­tone, insbesondere Malondialdehyd.The reaction of ozone with unsaturated fatty acids produces peroxidic products as therapeutically valuable active substances and low-molecular aldehydes and ketones, especially malondialdehyde, as undesirable by-products.

Die peroxidischen Produkte werden aufgrund ihrer keimtötenden Wirkung gegenüber Bakterien, Pil­zen und Hefen beispielsweise in der Dermatologie bei Pilzerkrankungen, Ulcus cruris, schlecht heilenden Wunden, infizierten Wunden und dergleichen eingesetzt. Demgegenüber sind die als Nebenprodukte gebildeten Ketone und Aldehyde therapeutisch ohne Wirkung und großenteils physiologisch bedenklich. Letzteres trifft insbesondere für Malondialdehyd zu, der bei vollständiger Ozonisierung von ungesättigten Pflan­zenölen in nicht unbeträchtlichen Mengen auftritt (vgl. Registry of Toxic Effects of Chemical Substan­ces, U.S. Department of Health and Human Services, 1984).Because of their germicidal action against bacteria, fungi and yeasts, the peroxidic products are used, for example, in dermatology for fungal diseases, leg ulcers, poorly healing wounds, infected wounds and the like. In contrast, the ketones and aldehydes formed as by-products are therapeutically ineffective and largely physiologically questionable. The latter is particularly true for malondialdehyde, which occurs in considerable quantities when ozonization of unsaturated vegetable oils is complete (see Registry of Toxic Effects of Chemical Substances, U.S. Department of Health and Human Services, 1984).

Aufgabe der Erfindung ist es, ozonisierte Pro­dukte aus ungesättigten Pflanzenölen bereitzustellen, die therapeutisch wertvolle Produkte darstellen, über lange Zeit hinweg stabil sind und einen mög­lichst geringen Gehalt an niedermolekularen Aldehy­den und Ketonen, insbesondere Malondialdehyd, auf­weisen.The object of the invention is to provide ozonized products from unsaturated vegetable oils which are therapeutically valuable products, are stable for a long time and have the lowest possible content of low molecular weight aldehydes and ketones, in particular malondialdehyde.

Im Verlauf von Untersuchungen, die zur Lösung dieser Aufgabe angestellt wurden, wurde festgestellt, daß sich Malondialdehyd aus Produkten, die durch Einleiten eines Ozon-Sauerstoff-Gemisches bis zur Sättigung des Produkts erhalten worden sind, leicht zu weit über 50 Prozent mit Wasser extrahieren läßt. Dieses Verfahren ist jedoch nicht praktikabel, da dabei die therapeutisch nutzbaren Peroxide zu etwa 90 Prozent zerfallen. Erfindungsgemäß wurde ein spe­zielles Extraktionsverfahren aufgefunden, das eine weitgehende Entfernung von Malondialdehyd ermöglicht, ohne daß es zu einer erheblichen Verminderung der Peroxidzahl, die ein Maß für den Gehalt an peroxi­dischen Produkten und somit für die Güte des Pro­dukts darstellt, kommt.In the course of investigations which were carried out to achieve this object, it was found that malondialdehyde can easily be extracted from water which has been obtained by introducing an ozone-oxygen mixture to saturation of the product to far more than 50 percent with water . However, this method is not practical since the peroxides which can be used therapeutically disintegrate to about 90 percent. According to the invention, a special extraction process has been found which enables extensive removal of malondialdehyde without significantly reducing the Peroxide number, which is a measure of the content of peroxidic products and thus of the quality of the product.

Gegenstand der Erfindung ist ein Verfahren zur Herstellung von stabilen ozonisierten Ölen aus ungesättigten Pflanzenölen durch Einleiten eines Ozon-Sauerstoff-Gemisches in das Öl bis zur Sätti­gung, das dadurch gekennzeichnet ist, daß man nach der Ozonisierung eine Extraktionsbehandlung in sau­rem Milieu in Gegenwart eines Redoxsystems, vorzugs­weise eines biologischen Redoxsystems, das zu einer gemäßigten radikalischen Reaktion fähig ist und in Gegenwart eines künstlichen oder natürlichen Anti­oxidationsmittels oder Reduktionsmittels durchführt. Als ungesättigte Pflanzenöle kommen beispielsweise Olivenöl, Distelöl, Weizenkeimöl, Leinöl, Mandelöl, Nußöl, Sonnenblumenöl, Mohnöl, Sesamöl, Rizinusöl, Krotonöl, Sojaöl und Palmöl in Frage. Oliven- und Distelöl werden bevorzugt. Ganz besonders bevorzugt wird Olivenöl.The invention relates to a process for producing stable ozonized oils from unsaturated vegetable oils by introducing an ozone-oxygen mixture into the oil until saturation, which is characterized in that after the ozonization, an extraction treatment in an acidic environment in the presence of a redox system, preferably a biological redox system which is capable of a moderate radical reaction and which carries out in the presence of an artificial or natural antioxidant or reducing agent. Examples of unsaturated vegetable oils are olive oil, safflower oil, wheat germ oil, linseed oil, almond oil, nut oil, sunflower oil, poppy seed oil, sesame oil, castor oil, croton oil, soybean oil and palm oil. Olive and safflower oils are preferred. Olive oil is especially preferred.

Das verwendete Redoxsystem, das zu radikali­schen Reaktionen fähig ist, wirkt im Organismus so­wohl als Elektronenakzeptor als auch als Elektronen­donator. Bevorzugte Beispiele für Redoxsysteme sind Ascorbinsäure, das im wesentlichen membranassoziierte Redoxsystem Vitamin E, Vitamin A und die biologi­schen chinoiden/benzoiden-Systeme. Besonders bevor­zugt wird Ascorbinsäure.The redox system used, which is capable of radical reactions, acts in the organism both as an electron acceptor and as an electron donor. Preferred examples of redox systems are ascorbic acid, the essentially membrane-associated redox system vitamin E, vitamin A and the biological quinoid / benzoid systems. Ascorbic acid is particularly preferred.

Zur Durchführung der Extraktionsbehandlung hat sich ein pH-Wert von 3,5 bis 6,5 als besonders geeignet erwiesen. Zur Gewährleistung dieses pH-Be­reichs können beispielsweise Ascorbinsäure oder Citronensäure eingesetzt werden.A pH of 3.5 to 6.5 has proven to be particularly suitable for carrying out the extraction treatment. To ensure this pH range, for example, ascorbic acid or citric acid can be used.

Bevorzugte Beispiele für Antioxidationsmittel bzw. Reduktionsmittel sind Butylhydroxyanisol, Gallat und Hydrogensulfit.Preferred examples of antioxidants or reducing agents are butylated hydroxyanisole, gallate and hydrogen sulfite.

Nachstehend wird die Erfindung unter Bezug­nahme auf Olivenöl näher erläutert. Entsprechende Ausführungen gelten auch für die Verwendung von Di­stelöl und anderen ungesättigten Pflanzenölen.The invention is explained in more detail below with reference to olive oil. Corresponding statements also apply to the use of safflower oil and other unsaturated vegetable oils.

Es wird reines Olivenöl entsprechend DAB 8 verwendet. Die Herstellung des Ozon-Sauerstoff-Gemi­sches erfolgt dabei unter Beachtung der Vorschriften zur Herstellung oder Abfüllung von medizinischem Sauerstoff. Das Ozon wird durch stille elektrische Entladung (absolut stickstofffrei, um die Bildung von aggressiven und reaktionsfähigen Stickoxiden, insbesondere Radikalen, zu vermeiden) gebildet. Bei der Ozonisierung des Öls wird mit einem kontinuier­lichen Durchlauf mit einem Gasfluß im Bereich von 1 bis 2 Liter pro Minute gearbeitet. Die verwendete Ozon-Konzentration liegt vorzugsweise im Bereich von 50 bis 70 µg pro ml. Die Durchperlung des Öls er­folgt unter Verwendung eines zylindrischen Gefäßes mit möglichst hoher "Wassersäule" unter Thermostati­sierung bei ca. 20°C. Dabei ist ein kontinuierlicher Durchfluß, insbesondere kurz vor der Sättigungsgren­ze, die sich durch Festwerden des Produkts bei 20°C bemerkbar macht, wichtig. Für 12 Liter bei einer Ge­samthöhe des zylindrischen Gefäßes von 60 cm und einem Durchmesser von ca. 20 cm ist eine gleichmäßi­ge Durchperlungszeit von 180 bis 300 Stunden je nach Ozonkonzentration des Ozon-Sauerstoff-Ausgangsgemi­sches erforderlich.Pure olive oil according to DAB 8 is used. The ozone-oxygen mixture is produced in compliance with the regulations for the production or filling of medical oxygen. The ozone is generated by a silent electrical discharge (absolutely nitrogen-free to avoid the formation of aggressive and reactive nitrogen oxides, especially radicals). The ozonization of the oil is carried out with a continuous pass with a gas flow in the range of 1 to 2 liters per minute. The ozone concentration used is preferably in the range from 50 to 70 μg per ml. The oil is bubbled through using a cylindrical vessel with the highest possible “water column” with thermostatting at about 20 ° C. A continuous flow is important, especially just before the saturation limit, which becomes apparent when the product solidifies at 20 ° C. For 12 liters with a total height of the cylindrical vessel of 60 cm and a diameter of approx. 20 cm, a uniform bubbling time of 180 to 300 hours is required, depending on the ozone concentration of the initial ozone-oxygen mixture.

Zur Extraktionsbehandlung werden pro 1 g ozo­nisiertes Öl etwa 2 bis 20, insbesondere 3 bis 10 und vorzugsweise etwa 5 ml wässrige Extraktionslö­sung verwendet. Die Extraktion erfolgt vorzugsweise durch eine 10- bis 60-minütige Ausschüttelung. Zur Steigerung der Effektivität kann der Ausschüttelvor­gang mehrfach durchgeführt werden. Im Extrakt even­tuell noch vorhandenes überschüssiges Sulfit kann durch Ausschütteln der öligen Phase mit 2-prozenti­ger wässriger Ascorbinsäurelösung entfernt werden.For the extraction treatment, about 2 to 20, in particular 3 to 10 and preferably about 5 ml of aqueous extraction solution are used per 1 g of ozonized oil. The extraction is preferably carried out by shaking for 10 to 60 minutes. The shaking process can be carried out several times to increase the effectiveness. Any excess sulfite still present in the extract can be removed by shaking out the oily phase with 2% aqueous ascorbic acid solution.

Maßgebend für die Qualität des Produkts sind insbesondere die Peroxidzahl, bestimmt gemäß Sully (DGF-Einheitsmethoden zur Untersuchung von Fetten, Fettprodukten und verwandten Stoffen, Deutsche Ge­sellschaft für Fettwissenschaft, Münster/Westfalen, 1950, S. 76) und der Gehalt an Malondialdehyd (be­stimmt gemäß H. Scherz et al., Mikrochem. Acta, Heft 5, 1967).Decisive for the quality of the product are in particular the peroxide number, determined according to Sully (DGF standard methods for the investigation of fats, fat products and related substances, German Society for Fat Science, Münster / Westphalia, 1950, p. 76) and the content of malondialdehyde (determined according to H. Scherz et al., Mikrochem. Acta, Issue 5, 1967).

Besonders bevorzugte Kombinationen für die erfindungsgemäße Extraktionsbehandlung sind Ascor­binsäure + Butylhydroxyanisol, Citronensäure und Butylhydroxyanisol sowie Ascorbinsäure und Hydrogen­sulfit.Particularly preferred combinations for the extraction treatment according to the invention are ascorbic acid + butylated hydroxyanisole, citric acid and butylated hydroxyanisole as well as ascorbic acid and hydrogen sulfite.

Nachstehend wird die Erfindung anhand von Bei­spielen näher erläutert.The invention is explained in more detail below with the aid of examples.

Bestimmung von MalondialdehydDetermination of malondialdehyde

Die Bestimmung beruht auf der Reaktion von Malondialdehyd mit 2-Methylindol in alkoholisch-salz­saurer Lösung zu einem beständigen, intensiv gefärb­ten Komplex.The determination is based on the reaction of malondialdehyde with 2-methylindole in alcoholic-hydrochloric acid solution to form a stable, intensely colored complex.

Reagentien und Standardlösungen:Reagents and standard solutions:

0,1 g 2-Methylenindol in 100 ml reinem EtOH gelöst; kurz vor der Verwendung 25 ml konz. HC1 zu­gesetzt.
Eichreihe: 45,86 mg Malondialdehydtetraäthylacetal werden mit 3 ml konz. HC1 versetzt und 3 min auf 50°C erwärmt. Danach wird mit destilliertem H₂O auf 500 ml ergänzt. 100, 200, 300, 400 und 500 µl dieser Lösung werden auf 1 ml aufgefüllt und mit 2 ml 2-Methylin­dollösung versetzt. Gleichzeitig wird ein Blindwert bestimmt, wobei 1 ml destilliertes H₂O mit 2 ml Rea­genslösung versetzt wird. Nach 20 min wird die Ex­tinktion bei 555 nm gemessen.0.1 g of 2-methylene indole dissolved in 100 ml of pure EtOH; shortly before use 25 ml conc. HC1 added.
Calibration series: 45.86 mg malondialdehyde tetraethyl acetal are mixed with 3 ml conc. HC1 added and heated to 50 ° C for 3 min. Then it is supplemented with distilled H₂O to 500 ml. 100, 200, 300, 400 and 500 µl of this solution are made up to 1 ml and 2 ml of 2-methanol solution are added. At the same time, a blank value is determined, 1 ml of distilled H₂O being mixed with 2 ml of reagent solution. After 20 min, the absorbance is measured at 555 nm.

Bestimmungen von Malondialdehyd im ozonisier­ten Öl: Etwa 1 g des festen Öls wird genau eingewo­gen und unter N₂-Atmosphäre zur Verflüssigung etwas erwärmt (ca. 30°C). Da das Öl mit den Reagentien nicht mischbar ist, werden außer 2 ml Methylindol noch 1 ml destilliertes H₂O zugesetzt und während der 20 min Reaktionszeit unter N₂-Atmosphäre mit einem Magnetrührer gerührt, um eine vollständige Reaktion des Malondialdehyds zu ermöglichen. Gemes­sen wird dann die alkoholische Phase (nach dem Ab­setzen des Fettes) im Vergleich mit der jedes Mal parallel angesetzten Eichreihe.Determination of malondialdehyde in ozonized oil: About 1 g of the solid oil is weighed exactly and warmed to liquefy under an N₂ atmosphere (approx. 30 ° C). Since the oil is immiscible with the reagents, 1 ml of distilled H₂O are added in addition to 2 ml of methyl indole and stirred with a magnetic stirrer under the N₂ atmosphere during the 20 min reaction time to allow a complete reaction of the malondialdehyde. The alcoholic phase is then measured (after the fat has been removed) in comparison with the calibration series that is applied in parallel each time.

Bestimmung der PeroxidzahlDetermination of the peroxide number

In einem trockenen, entfetteten Kolben mit aufgesetztem Kühler werden 10 ml Eisessig und 10 ml Chloroform zur Entlüftung der Mischung 2 min unter Rückfluß erhitzt, danach 1 kg KJ in 1,3 ml H₂O (frisch zubereitet) zugegeben, ohne das Sieden zu unterbrechen, und nach weiteren 2 min etwa 1 g Öl (genau eingewogen) zugesetzt. Nach 4 min werden 50 ml destilliertes H₂O zugegeben und auf Zimmertemperatur abgekühlt.In a dry, degreased flask with an attached cooler, 10 ml of glacial acetic acid and 10 ml of chloroform are vented for 2 minutes under reflux to vent the mixture, then 1 kg of KJ in 1.3 ml of H₂O (freshly prepared) are added without boiling interrupt, and after a further 2 minutes add about 1 g of oil (weighed exactly). After 4 min, 50 ml of distilled H₂O are added and the mixture is cooled to room temperature.

Nach Zugabe von 1 ml 1-prozentiger Stärkelö­sung werden unter Umschwenken mit 0,1 n Natriumsul­fatlösung bis zur Farblosigkeit der wässrigen Schicht titriert.After adding 1 ml of a 1% starch solution, titrate with 0.1 N sodium sulfate solution until the aqueous layer is colorless.

Berechnung:Calculation:

Figure imgb0001
Figure imgb0001

a ... verbrauchte ml Thiosulfatlösung
E ... Einwaage in ga ... used ml of thiosulfate solution
E ... weight in g

Beispiel 1example 1

12 Liter Olivenöl (gemäß DAB 8) werden in ein zylindrisches Gefäß von ca. 20 cm Durchmesser gege­ben. Die Füllhöhe beträgt ca. 60 cm. Das Gefäß wird auf 20°C thermostatisiert. Durch das GEfäß wird 240 Stunden lang gleichmäßig ein Ozon-Sauerstoff-Gemisch mit einer Durchlaufgeschwirdigkeit von 1 bis 2 Li­ter/min durchgeperlt. Dieses Ozon-Sauerstoff-Gemisch ist durch stille elektrische Entladung von reinem, stickstofffreiem Sauerstoff hergestellt worden. Seine Ozon-Konzentration beträgt ca. 60 µg/ml.12 liters of olive oil (according to DAB 8) are placed in a cylindrical vessel with a diameter of approx. 20 cm. The filling height is approx. 60 cm. The vessel is thermostatted to 20 ° C. An ozone-oxygen mixture with a flow rate of 1 to 2 liters / min is evenly bubbled through the vessel for 240 hours. This ozone-oxygen mixture was produced by the silent electrical discharge of pure, nitrogen-free oxygen. Its ozone concentration is approximately 60 µg / ml.

Das erhaltene Produkt weist eine Peroxidzahl (POZ) von 929 und einen Malondialdehyd-Gehalt von 445 µg/g Öl auf.The product obtained has a peroxide number (POZ) of 929 and a malondialdehyde content of 445 µg / g of oil.

100 g des ozonisierten Öls werden unter Stick­stoffatmosphäre unter Rühren auf 30 °C bis zur Ver­ flüssigung langsam entfernt und mit 500 ml einer 2-prozentigen Ascorbinsäurelösung in Gegenwart von 5 ml 38-prozentiger NaHSO₃-Lösung versetzt. Nach 15-minütigem Ausschütteln werden die Phasen getrennt und die ölige Phase erneut mit 300 ml einer 2-pro­zentigen Ascorbinsäurelösung ausgeschüttelt, um das überschüssige Sulfit vollständig zu entfernen. Bei erneuter Peroxidzahlbestimmung des Produkts ergibt sich ein Wertvron 876. Im Wässrigen Extrakt ergibt sich eine POZ von 0. Das ölige Produkt ehthält Malon­dialdehyd in einer Menge von 68 µg/g Öl und der wäss­rige Extract 424 µg/g Extraktionsmittel.100 g of the ozonated oil are stirred under a nitrogen atmosphere at 30 ° C. until Ver liquid slowly removed and mixed with 500 ml of a 2 percent ascorbic acid solution in the presence of 5 ml of 38 percent NaHSO₃ solution. After shaking for 15 minutes, the phases are separated and the oily phase is shaken again with 300 ml of a 2% ascorbic acid solution in order to completely remove the excess sulfite. When the peroxide number of the product is determined again, a value of 876 results. The POZ in the aqueous extract is 0. The oily product contains malondialdehyde in an amount of 68 µg / g oil and the aqueous extract contains 424 µg / g extractant.

Dies bedeutet, daß durch die Extraktionsbe­handlung der Malondialdehyd zu 85 Prozent entfernt worden ist, während sich die Peroxidzahl nur um 5,7 Prozent verrigerte. Der wässrige Extrakt enthält, wie gewünscht, keine Peroxide, während der Malon­dialdehyd fast vollständig in die wässrige Phase übergegangen ist.This means that the extraction treatment removed 85 percent of the malondialdehyde, while the peroxide number only decreased by 5.7 percent. As desired, the aqueous extract contains no peroxides, while the malondialdehyde has almost completely passed into the aqueous phase.

Beispiel 2Example 2

Proben des gemäß Beispiel 1 hergestellten ozonisierten Olivenöls werden unterschiedlichen Ex­traktionsbehandlungen unterworfen. Hierzu wird je­weils 1 g ozonisiertes Öl mit verschiedenen wässri­gen Lösungen 15 Minuten lang ausgeschüttelt. An­schließend werden POZ und Malondialdehydgehalt er­mittelt. Die Ergebnisse sind in nachstehender Ta­belle zusammengestellt.

Figure imgb0002
Samples of the ozonated olive oil produced according to Example 1 are subjected to different extraction treatments. For this purpose, 1 g ozonized oil is shaken out with various aqueous solutions for 15 minutes. The POZ and malondialdehyde content are then determined. The results are summarized in the table below.
Figure imgb0002

Sämtliche hier angeführten Werte sind Mittel­werte aus 2 Bestimmungen.
x)1 bedeutet, daß der betreffende Parameter nicht be­stimmt wurde,
x)2 überschüssiges Sulfit wurde durch einmaliges Aus­schütteln des Öls mit 10 ml H₂O (2 g Ascorbin­säure/100 ml) vollständig entfernt, wobei sich POZ und Malondialdehydgehalt des ozonisierten Öls nicht wesentlich veränderten: die Peroxid­zahl sank von 876 auf 860, der Malondialdehydge­halt von 68 µg/g auf 52 µg/g.
X ButylhydroxyanisolAll values listed here are mean values from two determinations.
x) 1 means that the parameter in question has not been determined,
x) 2 excess sulfite was completely removed by shaking the oil with 10 ml H₂O (2 g ascorbic acid / 100 ml), whereby the POZ and malondialdehyde content of the ozonated oil did not change significantly: the peroxide number decreased from 876 to 860, the malondialdehyde content from 68 µg / g to 52 µg / g.
X butylated hydroxyanisole

Besonders günstige Ergebnisse werden demnach durch Kombination von Ascorbinsäure und Butylhydro­xyanisol, Citronensäure und Butylhydroxyanisol sowie Ascorbinsäure und Hydrogensulfit erhalten.

Figure imgb0003
Accordingly, particularly favorable results are obtained by combining ascorbic acid and butylated hydroxyanisole, citric acid and butylated hydroxyanisole as well as ascorbic acid and bisulfite.
Figure imgb0003

Claims (7)

1. Verfahren zur Herstellung von stabilen ozoni­sierten Ölen aus ungesättigten Pflanzenölen durch Einleiten eines Ozon-Sauerstoff-Gemisches in das Öl bis zur Sättigung, dadurch gekennzeichnet, daß man nach der Ozonisierung eine Extraktionsbe­handlung in saurem Milieu in Gegenwart eines Redox­systems, das zu einer radikalischen Reaktion fähig ist und in Gegenwart eines künstlichen oder natürli­chen Antioxidationsmittels oder Reduktionsmittels durchführt.1. A process for the preparation of stable ozonized oils from unsaturated vegetable oils by introducing an ozone-oxygen mixture into the oil until saturation, characterized in that after the ozonization, an extraction treatment in an acidic environment in the presence of a redox system which leads to a radical reaction capable and performs in the presence of an artificial or natural antioxidant or reducing agent. 2. Verfahren nach Anspruch 1, dadurch ge­kennzeichnet, daß man als ungesättigtes pflanzliches Öl Olivenöl, Distelöl, Weizenkeimöl, Leinöl, Mandelöl, Nußöl, Sonnenblumenöl, Mohnöl, Sesamöl, Rizinusöl, Krotonöl, Sojaöl oder Palmöl verwendet.2. The method according to claim 1, characterized in that olive oil, safflower oil, wheat germ oil, linseed oil, almond oil, nut oil, sunflower oil, poppy oil, sesame oil, castor oil, croton oil, soybean oil or palm oil are used as the unsaturated vegetable oil. 3. Verfahren nach Anspruch 1 oder 2, dadurch gekennzeichnet, daß man als Redoxsystem Ascorbinsäure, Vitamin A, Vitamin E oder chinoide/­benzoide Systeme verwendet.3. The method according to claim 1 or 2, characterized in that ascorbic acid, vitamin A, vitamin E or quinoid / benzoid systems are used as the redox system. 4. Verfahren nach einem der vorstehenden Ansprü­che, dadurch gekennzeichnet, daß man die Extraktion bei einem pH-Wert von 3,5 bis 6,5 durch­führt.4. The method according to any one of the preceding claims, characterized in that one carries out the extraction at a pH of 3.5 to 6.5. 5. Verfahren nach einem der vorstehenden Ansprü­che, dadurch gekennzeichnet, daß man zur Gewährleistung des sauren Milieus Ascorbinsäure oder Citronensäure verwendet.5. The method according to any one of the preceding claims, characterized in that ascorbic acid or citric acid is used to ensure the acidic environment. 6. Verfahren nach einem der vorstehenden Ansprü­che, dadurch gekennzeichnet, daß man als Antioxidationsmittel Butylhydroxyanisol oder Gallat bzw. biologische Antioxidantien verwendet.6. The method according to any one of the preceding claims, characterized in that butylhydroxyanisole or gallate or biological antioxidants are used as antioxidants. 7. Verfahren nach einem der Ansprüche 1 bis 5, dadurch gekennzeichnet, daß man als Re­duktionsmittel Hydrogensulfit verwendet.7. The method according to any one of claims 1 to 5, characterized in that hydrogen sulfite is used as the reducing agent.
EP87100725A 1986-03-01 1987-01-20 Process for preparing stable ozonized oils from unsaturated vegetable oils Expired EP0235528B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AT87100725T ATE42965T1 (en) 1986-03-01 1987-01-20 PROCESS FOR THE PRODUCTION OF STABLE OZONIZED OILS FROM UNSATURATED VEGETABLE OILS.

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE3606735A DE3606735C2 (en) 1986-03-01 1986-03-01 Process for the production of stable ozonized oils from unsaturated vegetable oils
DE3606735 1986-03-01

Publications (2)

Publication Number Publication Date
EP0235528A1 true EP0235528A1 (en) 1987-09-09
EP0235528B1 EP0235528B1 (en) 1989-05-10

Family

ID=6295281

Family Applications (1)

Application Number Title Priority Date Filing Date
EP87100725A Expired EP0235528B1 (en) 1986-03-01 1987-01-20 Process for preparing stable ozonized oils from unsaturated vegetable oils

Country Status (3)

Country Link
EP (1) EP0235528B1 (en)
AT (1) ATE42965T1 (en)
DE (1) DE3606735C2 (en)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996025409A1 (en) * 1995-02-16 1996-08-22 Cosmart Ag Cosmetic or pharmaceutical compositions containing an ozonide-enriched natural vegetable oil, and uses thereof
FR2784388A1 (en) * 1998-10-07 2000-04-14 Bernard Sonnois Ozonized oil obtained using pure oxygen, useful as cellular regeneration promoter in cosmetic or pharmaceutical applications, e.g. for treating skin, joint or circulatory disorders
WO2000078699A1 (en) * 1999-06-18 2000-12-28 E.I. Du Pont De Nemours Oxidative cleavage of unsaturated oils and products obtained therefrom
WO2001037829A1 (en) * 1999-11-25 2001-05-31 Gomez Moraleda Manuel Antonio Composition comprising ozonized oils and/or other ozonized natural and/or synthetic products and their use in pharmaceutical, cosmetic, dietetic or food supplement compositions in human and veterinary medicine
WO2006021924A1 (en) * 2004-08-26 2006-03-02 Pieter Josua Le Roux Therapeutic oil from plant origine
EP1681338A1 (en) * 2003-10-02 2006-07-19 Sun Care Fuels Corporation Modifier for biodiesel fuel, fuel, methods relating to those
WO2007046122A2 (en) * 2005-10-17 2007-04-26 Bioo3Pharma S.R.L. Ozonised oil, process for the preparation and use thereof in medical field
WO2010039064A1 (en) * 2008-10-03 2010-04-08 Sca Hygiene Products Ab Wipe with odour control substance
RU2462271C1 (en) * 2008-10-03 2012-09-27 Ска Хайджин Продактс Аб Deodorising wet wipes
ITUB20155250A1 (en) * 2015-11-24 2017-05-24 Apuzzo Dario New nutraceutical composition, orally administered, useful for exercising a powerful stimulation of the metabolism, in particular of the muscular energy metabolism.
IT202000024967A1 (en) * 2020-10-23 2022-04-23 Giovanni Sandomenico OZONIZED FLUID FOR PROTECTION - DEFENSE OF THE FIRST RESPIRATORY TRACT

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE10157260B4 (en) * 2001-11-22 2005-12-15 Pharmoxid Arznei Gmbh + Co. Kg Spreadable ointment with ozonides and peroxides of fatty acids or their esters
DE102012007239A1 (en) 2012-04-10 2013-10-10 Wolfgang Winkelmann A pharmaceutical composition containing an oxygenated unsaturated fatty acid and an organic solvent
DE102015001289A1 (en) 2015-01-29 2016-08-04 Wolfgang Winkelmann Pharmaceutical compositions and pesticides containing oxygenated vegetable oils or fatty acids

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US2083572A (en) * 1928-04-25 1937-06-15 Susan M Mckee Process of treating oil
DE1068251B (en) * 1959-11-05 Chemische Fabrik Stockhausen S. Oie, Krefeld Process for the production of water-soluble or emulsifiable, electrolyte-resistant lubricants

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5869207A (en) * 1981-10-21 1983-04-25 Nippon Petrochem Co Ltd Purification of epoxidized product
DE3373675D1 (en) * 1983-01-06 1987-10-22 Lepeltier Marie Francoise Process for the preparation of peroxydated fats by biological low energy transformation, products obtained by this process, mixtures of these products with additives and new medicines made up of these products

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE1068251B (en) * 1959-11-05 Chemische Fabrik Stockhausen S. Oie, Krefeld Process for the production of water-soluble or emulsifiable, electrolyte-resistant lubricants
US2083572A (en) * 1928-04-25 1937-06-15 Susan M Mckee Process of treating oil

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996025409A1 (en) * 1995-02-16 1996-08-22 Cosmart Ag Cosmetic or pharmaceutical compositions containing an ozonide-enriched natural vegetable oil, and uses thereof
FR2730636A1 (en) * 1995-02-16 1996-08-23 Cosmart Ag NATURAL VEGETABLE OIL ENRICHED IN OZONIDES, COSMETIC OR PHARMACEUTICAL COMPOSITIONS CONTAINING THE SAME AND USES THEREOF
FR2784388A1 (en) * 1998-10-07 2000-04-14 Bernard Sonnois Ozonized oil obtained using pure oxygen, useful as cellular regeneration promoter in cosmetic or pharmaceutical applications, e.g. for treating skin, joint or circulatory disorders
WO2000078699A1 (en) * 1999-06-18 2000-12-28 E.I. Du Pont De Nemours Oxidative cleavage of unsaturated oils and products obtained therefrom
US6768029B1 (en) 1999-06-18 2004-07-27 Cambridge Biopolymers Limited Oxidative cleavage of unsaturated oils and products obtained therefrom
WO2001037829A1 (en) * 1999-11-25 2001-05-31 Gomez Moraleda Manuel Antonio Composition comprising ozonized oils and/or other ozonized natural and/or synthetic products and their use in pharmaceutical, cosmetic, dietetic or food supplement compositions in human and veterinary medicine
ES2162586A1 (en) * 1999-11-25 2001-12-16 Moraleda Manuel Gomez Composition comprising ozonized oils and/or other natural and/or synthetic ozonized products and use thereof in pharmaceutical, cosmetic, dietetic or food supplement compositions, in the human and veterinary fields
EP1681338A1 (en) * 2003-10-02 2006-07-19 Sun Care Fuels Corporation Modifier for biodiesel fuel, fuel, methods relating to those
EP1681338A4 (en) * 2003-10-02 2008-11-19 Sun Care Fuels Corp Modifier for biodiesel fuel, fuel, methods relating to those
WO2006021924A1 (en) * 2004-08-26 2006-03-02 Pieter Josua Le Roux Therapeutic oil from plant origine
WO2007046122A2 (en) * 2005-10-17 2007-04-26 Bioo3Pharma S.R.L. Ozonised oil, process for the preparation and use thereof in medical field
WO2007046122A3 (en) * 2005-10-17 2007-06-14 Biopharma 3M S R L Ozonised oil, process for the preparation and use thereof in medical field
WO2010039064A1 (en) * 2008-10-03 2010-04-08 Sca Hygiene Products Ab Wipe with odour control substance
RU2462271C1 (en) * 2008-10-03 2012-09-27 Ска Хайджин Продактс Аб Deodorising wet wipes
ITUB20155250A1 (en) * 2015-11-24 2017-05-24 Apuzzo Dario New nutraceutical composition, orally administered, useful for exercising a powerful stimulation of the metabolism, in particular of the muscular energy metabolism.
IT202000024967A1 (en) * 2020-10-23 2022-04-23 Giovanni Sandomenico OZONIZED FLUID FOR PROTECTION - DEFENSE OF THE FIRST RESPIRATORY TRACT

Also Published As

Publication number Publication date
ATE42965T1 (en) 1989-05-15
DE3606735C2 (en) 1994-11-10
EP0235528B1 (en) 1989-05-10
DE3606735A1 (en) 1987-09-10

Similar Documents

Publication Publication Date Title
EP0235528B1 (en) Process for preparing stable ozonized oils from unsaturated vegetable oils
DE3874428T2 (en) MULBERRY TREE EXTRACTS CONTAINING LIPOSOME COMPOSITION.
EP0060553B1 (en) Preparation for treating outer skin wounds and process for manufacturing said preparation
DE69807070T2 (en) TOCOFEROL ESTERN AND THEIR USE IN COSMETICS AND PHARMACEUTICAL
DE4210112A1 (en) POLYETHOXYLATED VITAMIN E AND METHOD FOR THE PRODUCTION THEREOF
DE1917393A1 (en) Perfumed and flavored products
DE2102172C3 (en) New means of treating and caring for the skin
DE3633453A1 (en) COMPOSITIONS BASED ON LYSINE AND ARGININE SALTS
DE2700448C2 (en) Ointment for the treatment of ulcus cruris
DE2129654B2 (en) Medicament for reducing capillary permeability and increasing capillary stability from the extraction of the flavanol-containing fruits of Vitis vinifera
AT414082B (en) PROCESS FOR THE PREPARATION OF TOCOTRIENOL-ENRICHED PREPARATIONS
EP0339382B1 (en) Process for deodorizing mixtures of fatty acid esters
DE69711633T2 (en) STABLE DERIVATIVES OF UBIQUINOL, METHOD FOR THE PRODUCTION THEREOF AND THEIR PHARMACEUTICAL USE
DE19728609C2 (en) Triglyceride-sorbic acid-pantothenyl-acetal peroxides from natural vegetable oils
DE3643323C2 (en)
EP0005121B1 (en) Process for stabilisation of 4-methyl-5-(2'-chlorethyl)-thiazol
DE1568320B2 (en) Cosmetic active ingredient and cosmetic agent with water-retaining properties
DE580515C (en) Process for the production of triaryl stib iodides and their solutions
EP0951904B1 (en) Agent for enhancing dermal microcirculation
DE2855860C2 (en) Process for the purification of organic peroxides containing water
DE2531260A1 (en) D-panthenol anhydrous spray solutions - contg an oil as solvent and an alcohol and a chlorinated hydrocarbon as solubilisers
DE60122696T2 (en) METHODS AND COMPOSITION FOR IMPROVED OXYGEN FEED FOR CELLS
DE2714342C2 (en) Bis (acetylsalicylic acid) thiaalkanediol ester and therapeutic composition
DE850054C (en) Toilet or shaving soap
DE624231C (en) Process for the production of hydrogenated tachysterin

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH FR IT LI NL

17P Request for examination filed

Effective date: 19871015

17Q First examination report despatched

Effective date: 19880713

GRAA (expected) grant

Free format text: ORIGINAL CODE: 0009210

AK Designated contracting states

Kind code of ref document: B1

Designated state(s): AT BE CH FR IT LI NL

REF Corresponds to:

Ref document number: 42965

Country of ref document: AT

Kind code of ref document: T

ET Fr: translation filed
ITF It: translation for a ep patent filed

Owner name: DR. ING. A. RACHELI & C.

PLBE No opposition filed within time limit

Free format text: ORIGINAL CODE: 0009261

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: NO OPPOSITION FILED WITHIN TIME LIMIT

26N No opposition filed
ITTA It: last paid annual fee
PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: FR

Payment date: 20031217

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: AT

Payment date: 20040116

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: BE

Payment date: 20040129

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: NL

Payment date: 20040130

Year of fee payment: 18

PGFP Annual fee paid to national office [announced via postgrant information from national office to epo]

Ref country code: CH

Payment date: 20040413

Year of fee payment: 18

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: IT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES;WARNING: LAPSES OF ITALIAN PATENTS WITH EFFECTIVE DATE BEFORE 2007 MAY HAVE OCCURRED AT ANY TIME BEFORE 2007. THE CORRECT EFFECTIVE DATE MAY BE DIFFERENT FROM THE ONE RECORDED.

Effective date: 20050120

Ref country code: AT

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050120

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: CH

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050131

Ref country code: LI

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050131

Ref country code: BE

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050131

BERE Be: lapsed

Owner name: *J. HANSLER G.M.B.H.

Effective date: 20050131

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: NL

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050801

REG Reference to a national code

Ref country code: CH

Ref legal event code: PL

PG25 Lapsed in a contracting state [announced via postgrant information from national office to epo]

Ref country code: FR

Free format text: LAPSE BECAUSE OF NON-PAYMENT OF DUE FEES

Effective date: 20050930

NLV4 Nl: lapsed or anulled due to non-payment of the annual fee

Effective date: 20050801

REG Reference to a national code

Ref country code: FR

Ref legal event code: ST

BERE Be: lapsed

Owner name: *J. HANSLER G.M.B.H.

Effective date: 20050131