Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-03-07
1997-01-28
Scheiner, Toni R.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 79, 435 792, 436 63, 552650, 552638, 552640, 552641, 552624, 552625, 552633, G01N 3353, G01N 33537, G01N 33543, G01N 3348
Patent
active
055977014
DESCRIPTION:
BRIEF SUMMARY
The invention relates to a process for determining androstenone contents in adipose tissues and to a device for carrying out this process.
5.alpha.-Androst-16-en-3-one, which is responsible for the sexual odor of the boar and has an intense urine-like odor, accumulates in the adipose tissue of the animals. This accumulation in the fat does not depend on the anatomical position of adipose tissue and affects equally, for example, subcutaneous fat (back fact), intermuscular fat, intramuscular fat and organ fat. This results in the considerable problem that meat from boars may cause, especially on heating during preparation, an odor of urine which is organoleptically extremely unpleasant. On the other hand, because of the anabolic effect of the hormones which are likewise, besides androstenone, produced in the testis, boars show considerable growth advantages. The approval of boar fattening within the EC demands a possible investigation in order to be able routinely to identify and cull those boars in which, because of advanced pubertal development, the deposition of sexual odor in the fat has already reached a very high level. Currently, the limit for the androstenone content which is still tolerable is suggested to be 0.5 .mu.g/g of adipose tissue.
Although reliable measurement processes, in particular with an enzyme-immunological bases, for determining androstenone in adipose tissue have already been disclosed to data, the known processes permit at the most laboratory investigations on random samples, but by no means continuous monitoring in parallel with the slaughter line because of the very long time required for the analyses.
It is an object of the invention to improve the known measurement processes so far that they can be carried out in a considerably shorter time and, if possible, can also be applied in a technically simple, automatable process.
This object is achieved according to the invention in a process of the type described at the outset by the determination process comprising the following steps:
a) liquefying an adipose tissue sample by heating at a predetermined temperature in the range from 45.degree. to 60.degree. C.;
b) mixing a defined amount of the liquid fat with a water-soluble solvent for the androstenone at the temperature of the liquid fat;
c) cooling the fat/solvent mixture to a predetermined temperature at which considerable proportions of the fat dissolved in the solvent on the one hand are separated out of the solution, and on the other hand however the predominant part of the androstenone dissolved in the solvent phase remains dissolved in the solvent;
d) removing a defined amount of the androstenone-containing solvent phase and diluting in a predetermined ratio with an aqueous buffer solution which is suitable for the detection process used;
e) measuring the androstenone content in the solvent/buffer solution phase by means of competitive immunological detection reactions known per se.
A particularly suitable detection process is represented by the measurement process which has been disclosed in the publication by R. Claus, G. Mahler and E. Munster in "ARCHIV FUR LEBENSMITTELHYGIENE", volume 39, number 4/1988, pages 87-90. Reference is hereby made to the entire contents of this description of the competitive immunological measurement process.
Said immunological measurement process is based on a defined amount of specific antibodies raised against the substance to be measured (in this case the androstenone) being initially present in a test system. For improved manipulation of the antibodies, they are immobilized on a receptacle plate, in particular a microtitre plate. The receptacle plate prepared in this way is incubated with an enzyme-labeled androstenone and with the solvent-containing buffer solution which contains the free androstenone to be determined. After the predetermined incubation time has elapsed, decantation is carried out and androstenone not bound to the antibodies is removed from the receptacle plate.
The proportion of enzyme-labeled androstenone present on t
REFERENCES:
Archiv Fur Lebensmittelhygiene, "Determination of the boar steroid 5a-androst-16-en-3-one in adipose tissue of pigs with a rapid microtitre plate enzyme-immunoassay (MTE)", vol. 39, No. 4, Jul., 1988, pp. 87-90.
Chemical Abstracts, Abstract No. 158023, "Quantitative determination of the boar taint substance 5.alpha-androst-16-en-3-one in fat", vol. 85, No. 21, Nov. 22, 1976, p. 386.
Chemical Abstracts, Abstract No. 55921, "A method for the rapid determination of the boar taint steroid androstenone [in the adipose tissue of swine]", vol. 111, No. 7, Aug. 14, 1989, 608.
Chemical Abstracts, Abstract No. 4049, "Method for determination of the odorous substance 5.alpha-androst-16-en-3-one in boar fat", vol. 83, No. 1, Jul. 7, 1975, p. 368.
Journal of Agricultural and Food Chemistry, "Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for C19-delta16-Steroids in Sera of Boar Pigs", vol. 38, No. 1, Jan. 1990, pp. 331-335.
Claus Rolf P.
Dehnhard Martin
Huff Sheela J.
Riedel-De Haen Aktiengesellschaft
Scheiner Toni R.
LandOfFree
Process for determining androstenone contents in adipose tissues does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Process for determining androstenone contents in adipose tissues, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for determining androstenone contents in adipose tissues will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-940780