摘要:
Compositions and methods for detecting the presence and/or amount of one or more analytes, including analytes such as drugs of abuse, are provided. The compositions include two or more analytes associated with a solid phase, e.g., a particle or a multiwell plate. The compositions and methods also allow the simultaneous, tandem, or serial determination of the presence and/or amount of two or more analytes of interest in a sample.
摘要:
A method for the direct analysis of analyte indicative of marijuana exposure found in keratinized structures, e.g., hair, fingernails and toenails, which comprises preparing a mixture containing dithiothreitol or dithioerythritol, a protease suitable for the digestion of the keratin structure and a sample of the keratin structure; permitting the enzyme to at least substantially digest the sample of keratin structure to form a digest solution, followed by mixing the digest solution with a suspension of an ion exchange resin to remove an interfering, cross reacting substance naturally found in hair and finally subjecting the digest solution to analysis to determine the identity and amount of marijuana analyte in the keratin structure sample. To accelerate the method, cupric sulfate may be added to the mixture after degradation of the keratin sample in order to deactivate the activator. The enzyme may be a protease with papain, chymopapain, and proteinase K being preferred for use in the invention. Exemplary ion exchange resins useful in the method according to the invention are DEAE Sephadex (Diethylaminoethyl Sephadex) and DEAE Sepharose (Diethylaminoethyl Sepharose).
摘要:
Une méthode d'analyse directe d'un analyte dans des structures kératinisées, par exemple les cheveux et les ongles, consiste à préparer un mélange contenant un composé activateur à faible potentiel redox tel que le dithiothréitol ou le dithioérythritol, une enzyme appropriée pour la digestion de la structure de kératine, un échantillon de la structure de kératine et un détergent biologique qui facilite la digestion de la structure kératinisée à un pH relativement faible, par exemple entre 6,2 et 8; on laisse ensuite l'enzyme digérer l'échantillon de la structure de kératine, et on soumet la solution de digestion à une analyse, de préférence par radio immuno-analyse afin de déterminer l'identité et la quantité de l'analyte dans l'échantillon de la structure de kératine. Pour accélérer le procédé, on peut ajouter le sulfate de cuivre au mélange après dégradation de l'échantillon de kératine. L'enzyme peut être une peptidase, un endopeptidase ou une protéinase, les enzymes utilisées de préférence dans l'invention étant la papaïne, la chimiopapaïne et la protéinase K. Les détergents biologiques préférés comprennent la bétaïne, la sulfo-bétaïne les alkylglucosides et les acides biliaires.
摘要:
Methods for assessing the condition of keratinized structures, including hair, in particular methods to determine the condition of keratinized structures in relation to suitability for analysis of analytes of interest in a test sample, are presented. The methods comprise contacting the keratinized structure with a non-proteolytic reducing agent and an optional proteolytic agent. The methods further include inspection of the hair sample, or measurement of free protein eluted from the keratinized structure, after reduction and optional proteolysis to determine condition prior to analyte identification and quantitation by known techniques such as immunoassays.
摘要:
A method the direct analysis of an analyte in keratinized structures, e.g., hair and fingernails, which comprises preparing a mixture containing a low redox potential activator compound such as dithiothreitol or dithioerythritol, an enzyme suitable for the digestion of the keratin structure, a sample of the keratin structure and a biological detergent that aids the digestion of the keratinized structure at a relatively low pH, e.g., between about 6.2 and 8; permitting the enzyme to at least substantially digest the sample of keratin structure, and subjecting the digest solution to analysis, preferably by radioimmunoassay, to determine the identity and amount of analyte in the keratin structure sample. To accelerate the method, cupric sulfate may be added to the mixture after degradation of the keratin sample. The enzyme may be a peptidase, endopeptidase or proteinase, with papain, chymopapain, and proteinase K being preferred for use in the invention. The preferred biological detergents include betaine, sulfo-betaine, alkylglucosides and bile acids.
摘要:
A method for the direct analysis of analyte in keratinized structures, e.g., hair, fingernails and toenails, which comprises preparing a mixture containing a low redox potential compound such as dithiothreitol or dithioerythritol, an enzyme suitable for the degradation of the keratine structure and a sample of the keratin structure; permitting the enzyme to at least substantially degrade the sample of keratin structure, and subjecting the mixture to analysis to determine the identity and amount of analyte in the keratin substance sample. To accelerate the method, cupric sulfate or sodium arsenite may be added to the mixture after degradation of the keratin sample. The enzyme may be a peptidase, endopeptidase or proteinase, with papain, chymopapain, and proteinase K being preferred for use in the invention.