Claims for Patent: 8,986,939

Summary for Patent: 8,986,939

Title:	Integrity testing of hair samples
Abstract:	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.
Inventor(s):	Hill; Virginia (Los Angeles, CA), Schaffer; Michael I. (Los Angeles, CA), Loni; Elvan (Torrance, CA), Stowe; Gary Neil (Manhattan Beach, CA)
Assignee:	Psychemedics Corporation (Acton, MA)
Application Number:	14/472,252
Patent Claims:	1. A method for determining the presence or absence of a drug of abuse or metabolite thereof in a hair sample of a subject comprising: providing a hair sample from the subject; contacting the hair sample with an aqueous solution comprising a reducing agent to result in a test sample; subjecting the aqueous portion of the test sample to denaturing conditions; measuring the absorption at a wavelength of about 300 nm (A.sub.300) to about 380 nm (A.sub.380) of the subjected aqueous portion and comparing the measured absorbance value to the absorbance value of a control sample to determine if the hair sample is suitable for further processing; and determining if an analyte is present or absent in a hair sample identified as suitable for further processing to determine the presence or absence of a drug of abuse or metabolite thereof in the hair sample. 2. The method of claim 1, wherein subjecting the aqueous portion of the test sample to denaturing conditions comprises adjusting the pH of the aqueous portion to a pH of about 4.0 or less. 3. The method of claim 1, wherein determining if the hair sample is suitable for further processing comprises estimating the degree of damage to hair in the test sample using said measured absorption of said subjected aqueous portion, wherein said measured absorption is measured at a wavelength of about 340 nm (A.sub.340) to about 380 nm (A.sub.380). 4. The method of claim 1, wherein determining if the hair sample is suitable for further processing comprises estimating the degree of damage to hair in the test sample using said measured absorption of said subjected aqueous portion, wherein said measured absorption is measured at a wavelength of about 380 nm (A.sub.380), wherein a subjected aqueous portion having an A.sub.380 of 0.35 or greater identifies the hair sample as not being suitable for further processing and wherein a subjected aqueous portion having an A.sub.380 of less than 0.35 identifies the hair sample as being suitable for further processing. 5. The method of claim 1, wherein subjecting the aqueous portion of the test sample to denaturing conditions comprises adding an organic solvent to the aqueous portion of the test sample, wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, 2-propanol, acetone, acetonitrile, and mixtures thereof. 6. The method of claim 1, wherein subjecting the aqueous portion of the test sample to denaturing conditions comprises adding trichloroacetic acid, acetic acid, or sulfosalicyc acid to the aqueous portion of the test sample. 7. The method of claim 1, wherein the reducing agent is selected from the group consisting of 2,3 dihydroxybutane-1,4-dithiol ("DTT"), 2,3 dihydroxybutane-1,4-dithiol ("DTE"), thioglycolate, cysteine, sulfites, bisulfites, sulfides, bisulfides, tris(2-carboxyethyl)phosphine ("TCEP") and mixtures thereof. 8. A method for determining the presence or absence of a drug of abuse or metabolite thereof in a hair sample comprising: providing a hair sample from a subject; determining if the hair sample is not suitable for drug analyte testing comprising: contacting the hair sample with an aqueous solution comprising a reducing agent to result in a test sample, wherein the contacting step occurs for a time period of about 1 hour to about 2 hours; and identifying the hair in the test sample as not suitable for drug analyte testing when the hair in the test sample has dissolved in the test sample more rapidly than the hair in a similarly contacted control sample or the hair in the test sample appears softer or less rigid as compared to the hair in the similarly contacted control sample; and if the hair in the test sample is not identified as not suitable for drug analyte testing, then determining if the drug of abuse is present or absent in the hair sample. 9. The method of claim 8, wherein the rate at which the hair dissolved in the test sample or a similarly contacted control sample, or the appearance of the hair dissolved in the test sample or a similarly contacted control sample is evaluated by visual inspection. 10. The method of claim 8, wherein the reducing agent is selected from the group consisting of 2,3 dihydroxybutane-1,4-dithiol ("DTT"), 2,3 dihydroxybutane-1,4-dithiol ("DTE"), thioglycolate, cysteine, sulfites, bisulfites, sulfides, bisulfides, tris(2-carboxyethyl)phosphine ("TCEP") and mixtures thereof. 11. The method of claim 8, wherein the aqueous solution comprises 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1.0% of the reducing agent. 12. The method of claim 8, wherein the pH at which the contacting step is performed is between about 7.0 and 10.5, between about 8.8 and 10.5, between about 8.8 and 9.7, between about 8.8 and 9.5, or between about 9.4 and 9.7. 13. The method of claim 8, wherein the temperature at which the contacting step or the determining step is performed is between about 20.degree. C. and about 40.degree. C. 14. The method of claim 8, further comprising indicating the hair sample as unsuitable for determining if an analyte is present in the test solution if the hair sample has been identified as damaged. 15. The method of claim 8, wherein determining the presence or absence of a drug of abuse or a metabolite thereof comprises an enzyme immunoassay specific for the analyte, a mass spectrometry technique, or a chromatographic technique. 16. The method of claim 8, wherein the drug of abuse or metabolite thereof is selected from the group consisting of a prescription medicine or metabolite thereof, a pain medication or metabolite thereof, a nutrient, and an endogenous analyte, or a salt form of any of the foregoing. 17. The method of claim 8, further comprising determining the amount of protein in the hair sample. 18. The method of claim 17, wherein the amount of protein in the hair sample is determined using a protein assay selected from the group consisting of the Lowry Assay or the Bradford Assay. 19. The method of claim 8, wherein the rate at which the hair dissolved in the test sample or a similarly contacted control sample, or the appearance of the hair dissolved in the test sample or a similarly contacted control sample is completed using a portion of the hair sample; and determining the presence or absence of a drug of abuse or metabolite thereof is completed using: (a) the same portion of the hair sample; or (b) another portion of the hair sample. 20. The method of claim 8, further comprising deactivating residual reducing agent present in the test solution prior to determining the presence or absence of a drug of abuse or metabolite thereof, wherein the deactivating does not proteolytically cleave the keratinized structure, to result in a deactivated test solution. 21. The method of claim 20, wherein the deactivation step comprises contacting the test solution with an aqueous solution of a metal salt, wherein the metal cation of the salt is selected from the group consisting of Cu++, Zn++, Mn++, Fe+++, Fe++, Pb++, Cd++, Hg++, Ag++, As+++, and Co++. 22. A method for determining the presence or absence of a drug of abuse or metabolite thereof in a hair sample of a subject comprising: providing a hair sample; determining if the hair sample is not suitable for drug analyte testing comprising: contacting the hair sample with an aqueous solution comprising a reducing agent and a protease capable of digesting keratin to result in a test sample, wherein the contacting step occurs for a time period of about 1 hour to about 2 hours; identifying the hair in the test sample as not suitable for drug analyte testing when the hair in the test sample has dissolved in the test sample more rapidly than the hair in a similarly contacted control sample or the hair in the test sample appears softer or less rigid as compared to the hair in the similarly contacted control sample; and if the hair in the test sample is not identified as not suitable for drug analyte testing, then determine if the drug of abuse is present or absent in the hair sample. 23. The method of claim 22, wherein the protease is selected from the group consisting of papain, chymopapain, and proteinase K. 24. The method of claim 22, wherein the rate the hair dissolved in the test sample or a similarly contacted control sample or the appearance of the hair dissolved in the test sample or a similarly contacted control sample is assessed by visual inspection at predetermined time intervals. 25. The method of claim 22, wherein the reducing agent is selected from the group consisting of 2,3 dihydroxybutane-1,4-dithiol ("DTT"), 2,3 dihydroxybutane-1,4-dithiol ("DTE"), thioglycolate, cysteine, sulfites, bisulfites, sulfides, bisulfides, tris(2-carboxyethyl)phosphine ("TCEP"), and mixtures thereof. 26. The method of claim 22, wherein the aqueous solution comprises 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9% or 1.0% of the reducing agent. 27. The method of claim 22, wherein the pH at which the contacting step is performed is between about 7.0 and 10.5, between about 8.8 and 10.5, between about 8.8 and 9.7, between about 8.8 and 9.5, or between about 9.4 and 9.7. 28. The method of claim 22, wherein the temperature at which the contacting step or the determining step is performed is between about 20.degree. C. and about 40.degree. C. 29. The method of claim 22, wherein determining the presence or absence of a drug of abuse or metabolite thereof comprises an enzyme immunoassay specific for the analyte, a mass spectrometry technique, or a chromatographic technique.

Details for Patent 8,986,939

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	11/10/1982		2033-08-28
Discure Medical, Llc	CHYMODIACTIN	chymopapain	For Injection	018663	08/21/1984		2033-08-28