Claims for Patent: 9,261,522

Summary for Patent: 9,261,522

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:	14/664,032
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; 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 period of about 1 hour to about 2 hours; determining the amount of dissolved hair protein in the test sample using a protein assay; 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; 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 or metabolite thereof is present or absent in the hair sample. 2. The method of claim 1, wherein the protease is selected from the group consisting of papain, chymopapain, and proteinase K. 3. The method of claim 1, further comprising assessing the condition of the hair in the test sample by visual inspection at predetermined time intervals. 4. 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. 5. The method of claim 1, 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. 6. The method of claim 1, 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. 7. The method of claim 1, 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. 8. The method of claim 1, wherein determining the amount of dissolved hair protein in the test sample occurs at about 15 minute intervals. 9. The method of claim 3, wherein the predetermined time intervals are about 15 minute intervals. 10. The method of claim 1, wherein the step of determining the amount of dissolved hair protein in the test 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. 11. The method of claim 1, further comprising identifying the hair sample as being previously treated with potassium hydroxide if the hair sample fails to dissolve after a time period of about 120 minutes. 12. The method of claim 1, further comprising identifying the hair sample as being previously treated with a cellophane hair treatment if a dye is visible in the test solution within about 5 minutes of contacting the hair sample with an aqueous solution comprising a reducing agent and a protease. 13. The method of claim 1, wherein the hair sample is a washed hair sample. 14. The method of claim 1, wherein the amount of dissolved hair protein the test sample is determined using a protein assay selected from the group consisting of the Lowry Assay or the Bradford Assay. 15. The method of claim 1, 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. 16. The method of claim 1, 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. 17. The method of claim 1, 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. 18. The method of claim 17, wherein the drug of abuse or metabolite thereof is selected from the group consisting of cocaine, benzoylecgonine, cocaethylene, norcocaine, phencyclidine (PCP), amphetamine, methamphetamine, cannabinoid, THC, carboxy-THC, heroin, codeine, morphine, 6-monoacetylmorphine (MAM), oxycodone, 3,4-methylenedioxyamphetamine (MDA) and 3,4-methylenedioxymethamphetamine (MDMA). 19. The method of claim 17, wherein the prescription medicine or metabolite thereof, or pain medication or metabolite thereof is an opioid, cannabinoid, nonsteroid anti-inflammatory drugs (NSAID), steroid, amphetamine, benzodiazepine, barbiturate, tricyclic, or ephedrine, or metabolite thereof. 20. The method of claim 1, further comprising determining the amount of the analyte in the test solution. 21. The method of claim 1, wherein determining the amount of dissolved hair protein in the test sample occurs at 0, 15, 30, 45, 60, 75, 90 and 120 minutes after contacting the hair sample with an aqueous solution comprising a reducing agent and a protease capable of digesting keratin. 22. The method of claim 1, further comprising identifying the hair sample as suitable for further processing if the rate of dissolution in the test solution is commensurate with the rate of dissolution in the control sample. 23. The method of claim 1, further comprising determining the rate of dissolution of the hair in the test sample using a protein assay; 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. 24. 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; 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 period of about 1 hour to about 2 hours; determining the rate of dissolution of the hair in the test sample using a protein assay; 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; 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 or metabolite thereof is present or absent in the hair sample.

Details for Patent 9,261,522

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