Method for the detection of proteins of animal origin in complex mixtures
First Claim
1. Method for detecting proteins of animal origin in complex mixtures comprising the following stages:
- (i) extraction of proteic material in high concentration from a sample of the initial complex mixture in a manner as to substantially remove all interferents;
(ii) preparation of the matrix-analyte in a manner as to maintain low levels of impurities and an adequate matrix-analyte molar ratio;
(iii) analysis of the material obtained in the prior stage by MALDI-TOF mass spectrometry;
(iv) optionally, fractionation of the samples or isolation of the components by RP-HPLC and identification of the components by means of automatic sequencing of the N-terminal region and sequencing of its peptidic fragments by liquid chromatography coupled to mass spectrometry (LC/MS/MS).
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Abstract
The purpose of the present invention is to evaluate the quality of feed for ruminants and consequently, avoid the transmission of TSEs through the detection of animal proteins in these foods. This purpose is embodied in the form of a method for detecting proteins of animal origin in complex mixtures comprising the stages of: (i) extraction of the proteic matter in high concentration from a sample of the initial complex mixture in a manner as to substantially remove all interferents; (ii) preparation of the matrix-analyte in a manner as to maintain low levels of impurities and an adequate matrix-analyte molar rate; (iii) analysis of the material obtained in the prior stage by MALDI-TOF mass spectrometry; (iv) optionally, fractionation of the samples or isolation of the components by RP-HPLC and identification of the components by means of automatic sequencing of the N-terminal region and sequencing of its peptidic fragments by liquid chromatography coupled to mass spectrometry (LC/MS/MS). The present invention also contemplates the use of this method in the detection of proteins of animal origin in feed for ruminants, which permits the interruption of transmission of Transmittable Spongiform Encephalopathies, and more particularly Bovine Spongiform Encephalopathies (BSE).
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Citations
12 Claims
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1. Method for detecting proteins of animal origin in complex mixtures comprising the following stages:
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(i) extraction of proteic material in high concentration from a sample of the initial complex mixture in a manner as to substantially remove all interferents;
(ii) preparation of the matrix-analyte in a manner as to maintain low levels of impurities and an adequate matrix-analyte molar ratio;
(iii) analysis of the material obtained in the prior stage by MALDI-TOF mass spectrometry;
(iv) optionally, fractionation of the samples or isolation of the components by RP-HPLC and identification of the components by means of automatic sequencing of the N-terminal region and sequencing of its peptidic fragments by liquid chromatography coupled to mass spectrometry (LC/MS/MS). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification