Oral detection test for cannabinoid use

• US 7,465,586 B2
• Filed: 11/03/2004
• Issued: 12/16/2008
• Est. Priority Date: 11/03/2004
• Status: Active Grant

• Current Assignee: Labone Incorporated (Quest Diagnostics Incorporated)
• Original Assignee: Labone Incorporated (Quest Diagnostics Incorporated)
• Inventors: Day, David, Kuntz, David J., Feldman, Michael S.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US10/980,372
• Publication Number:
  US 20060094123A1
• Time in Patent Office: 1,504 Days
• Field of Search: 436/63, 436/161, 436/173, 436/169, 436/901, 436/93
• US Class Current: 436/93
• CPC Class Codes: G01N 33/948 Sedatives, e.g. cannabinoids, barbiturates opiates G01N33/9486
• CPC Class Codes: Y10S 436/901 Drugs of abuse, e.g. narcotics, amphetamine
• CPC Class Codes: Y10T 436/142222 Hetero-O [e.g., ascorbic acid, etc.]
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

A method for confirming the active intake of marijuana and its active component Δ⁹-tetrahydrocannabinol (“Δ⁹-THC”) by detecting the amount of 11-nor-Δ⁹-THC carboxylic acid (“THCA”) in oral fluid at the picogram per milliliter (pg/ml) level using chromatography/mass spectrometry/mass spectrometry (“GC/MS/MS”).

Claims

1. A method for confirming the active intake of marijuana in a test subject comprising the steps of:

obtaining a test sample of oral fluid from said test subject; and

detecting the amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in said test sample,

wherein said THCA has not been externally added to said test sample and said amount of THCA is less than about 240 pg/ml but greater than a predetermined cutoff amount;

wherein the presence of THCA in said test sample in an amount greater than said predetermined cutoff amount confirms the active intake of marijuana by said test subject.

2. The method of claim 1 wherein said test sample is obtained from said test subject using a collection pad which is placed in the oral cavity of said test subject for about 30 seconds to about five minutes.

3. The method of claim 2 wherein said obtaining step is performed by placing an absorbent pad in the oral cavity of said test subject for up to about five minutes.

4. The method of claim 3 wherein said pad is impregnated with one or more of a preservative, protein binding blocking agent, or detergent.

5. The method of claim 3 further comprising the step of storing the test sample in a preservative solution prior to said detecting step.

6. The method of claim 3 further comprising the step of extracting THCA and/or THCA-glucuronide from the absorbent pad using one or more organic reagents.

7. The method of claim 6 wherein said organic reagents are selected from the group consisting of hexane, acetic acid, formic acid, trifluoroacetic acid, dichioromethane, chloroform, acetonitrile, methanol, and ethanol.

8. The method of claim 3 further comprising the step of extracting THCA and/or THCA-glucuronide from the pad using an aqueous reagent mixture.

9. The method of claim 8 wherein said aqueous reagent mixture comprises water plus one or more agents selected from the group consisting of zinc chloride, magnesium chloride, sodium sulfide, sodium deoxycholate, sodium hydroxide, and potassium hydroxide.

10. The method of claim 1 further comprising the step of liberating THCA from THCA-glucuronide in the oral fluid prior to said detecting step.

11. The method of claim 10 wherein said liberating step is performed by exposing said oral fluid to alkaline conditions.

12. The method of claim 1 further comprising the step of concentrating the amount of THCA in the test sample prior to said detecting step, wherein said concentrating step comprises passing said test sample through a solid phase extraction column.

13. The method of claim 1 wherein the step of detecting of the amount THCA in said test sample is performed using gas chromatography/mass spectrometry/mass spectrometry.

14. The method of claim 1 wherein the predetermined cutoff amount of THCA is at least about 20 pg/ml.

15. The method of claim 1 wherein the predetermined cutoff amount of THCA is about 10 pg/ml.

16. The method of claim 1 further comprising the step of comparing the detected amount of THCA in said test sample with a control amount of THCA in a control sample of oral fluid from a control test subject who has not actively consumed marijuana, and wherein said detected amount of THCA in said test sample is higher than said control amount of THCA in said control sample by a predetermined differential to indicate the active intake of marijuana by said test subject.

17. The method of claim 16 wherein the predetermined differential is about 10 pg/ml or more.

18. The method of claim 16 wherein the predetermined differential is about 20 pg/ml or more.

19. A method for testing whether a test subject has falsely tested negative for the active intake of marijuana comprising the steps of:

obtaining a test sample of oral fluid from said test subject who has tested negative for active marijuana intake; and

detecting the amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in said oral fluid in said test sample, wherein said THCA has not been externally added to said test sample and said amount of THCA is less than about 240 pg/ml but greater than a predetermined cutoff amount;

wherein the presence of THCA in said test sample in an amount greater than said predetermined cutoff amount indicates the active intake of marijuana by said test subject.

20. The method of claim 19 further comprising the step of comparing the detected amount of THCA in said test sample with a control amount of THCA in a control sample of oral fluid from a control subject who has not actively consumed marijuana, and wherein said detected amount of THCA in said test sample is higher than said control amount of THCA in said control sample by a predetermined differential to indicate the active intake of marijuana by said test subject.

21. The method of claim 20 wherein the predetermined differential is about 10 pg/ml or more.

22. The method of claim 20 wherein the predetermined differential is about 20 pg/ml or more.

23. The method of claim 19, wherein said predetermined cutoff amount of THCA is about 10 pg/ml.

24. The method of claim 19, wherein said predetermined amount of THCA is about 20 pg/ml or more.

33. The method of claim 22, wherein said aqueous reagent mixture comprises water plus one or more agents selected from the group consisting of zinc chloride, magnesium chloride, sodium sulfide, sodium deoxycholate, sodium hydroxide, and potassium hydroxide.

25. A method for determining active intake of marijuana by a subject, comprising:

(i) detecting by an assay an amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in an oral fluid sample from said subject, wherein said assay is capable of detecting THCA in the pg/ml range; and

(ii) comparing said detected amount of THCA with a cutoff value wherein said cutoff value is a value between about 10 pg/ml and about 30 pg/ml,

wherein when said amount of THCA detected is higher than said cutoff value, active intake of marijuana by said subject is indicated.

26. The method of claim 25, wherein said cutoff value is between about 20 pg/ml and about 30 pg/ml.

27. The method of claim 25, wherein said sample is obtained from said subject by placing a collection pad in the oral cavity of said subject for about 30 seconds to about five minutes.

28. The method of claim 27, wherein said pad is impregnated with one or more of a preservative, a protein binding blocking agent, and a detergent.

29. The method of claim 27, further comprising the step of storing said sample in a preservative solution prior to said detecting step.

30. The method of claim 27, further comprising the step of extracting THCA and/or THCA-glucuronide from said pad using one or more organic reagents.

31. The method of claim 30, wherein said organic reagents are selected from the group consisting of hexane, acetic acid, formic acid, trifluoroacetic acid, dichloromethane, chloroform, acetonitrile, methanol, and ethanol.

32. The method of claim 27, further comprising the step of extracting THCA and/or THCA-glucuronide from said pad using an aqueous reagent mixture.

34. The method of claim 25, further comprising the step of liberating THCA from THCA-glucuronide in the oral fluid prior to said detecting step.

35. The method of claim 34, wherein said liberating step is performed by exposing said oral fluid to alkaline conditions.

36. The method of claim 35, further comprising the step of concentrating the amount of THCA in the sample prior to said detecting step, wherein said concentrating step comprises passing said sample through a solid phase extraction column.

37. The method of claim 25, wherein the amount of THCA detected is between about 10 pg/ml and less than about 240 pg/ml.

38. A method for determining active intake of marijuana by a subject, comprising:

(i) detecting by an assay an amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in an oral fluid sample from said subject, wherein said assay is capable of detecting THCA in the pg/ml range; and

(ii) comparing said detected amount of THCA with a control amount of THCA in a control sample of oral fluid from a control subject, who has not actively consumed marijuana, wherein when said amount of THCA detected is higher than said control amount of THCA by a preset differential of between about 10 pg/ml and about 240 pg/ml, active intake of marijuana by said subject is indicated.

39. The method of claim 38, wherein said preset differential is between about 20 pg/ml and about 240 pg/ml.

40. The method of claim 38, wherein said assay is mass spectrometry.

41. The method of claim 38, wherein the amount of THCA detected is between about 10 pg/ml and less than about 240 pg/ml.

42. A method for determining whether a subject has falsely tested negative for active intake of marijuana, comprising:

(i) detecting by an assay an amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in an oral fluid sample from said subject, wherein said assay is capable of detecting THCA in the pg/ml range; and

(ii) comparing said detected amount of THCA with a control amount of THCA in a control sample of oral fluid from a control subject, who has not actively consumed marijuana, wherein when said detected amount of THCA is higher than said control amount of THCA by a preset differential of between about 10 pg/ml and about 240 pg/ml said subject falsely tested negative for active intake of marijuana is indicated.

43. The method of claim 42, wherein said preset differential is between about 20 pg/ml and about 240 pg/ml.

44. The method of claim 42, wherein said assay is mass spectrometry.

45. The method of claim 42, wherein the amount of THCA detected is between about 10 pg/ml and less than about 240 pg/ml.

46. A method for determining active intake of marijuana by a subject, comprising:

(i) detecting a specified amount of 11-nor-Δ

⁹ THC carboxylic acid (“

THCA”

) in an oral fluid sample from said subject by mass spectrometry, and

(ii) comparing said detected amount of THCA with a cutoff value wherein said cutoff value is a value between about 10 pg/ml and about 30 pg/ml,

wherein when said amount of THCA detected is higher than said cutoff value, active intake of marijuana by said subject is indicated; and

wherein said mass spectrometry is capable of detecting the amount of THCA in said sample in the pg/ml range.

47. The method of claim 46, wherein said cutoff value is determined relative to the amount of THCA in a control subject.

48. The method of claim 46, wherein said cutoff value is between about 20 pg/ml and about 30 pg/ml.

49. The method of claim 46, wherein the amount of THCA detected is between about 10 pg/ml and less than about 240 pg/ml.

50. The method of claim 46, wherein the mass spectrometry is performed by a mass spectrometer comprising a mass analyzer, wherein said mass analyzer is selected from the group consisting of a quadrupole mass filter, an ion trap mass analyzer, a time-of-flight mass analyzer, a magnetic sector, and a Fourier transform ion cyclotron resonance spectrometer.

51. The method of claim 50, wherein the mass analyzer is capable of detecting the amount of THCA in said sample in the range between 10 pg/ml and 240 pg/ml.

52. The method of claim 46, wherein the mass spectrometry is tandem mass spectrometry.

53. The method of claim 46, wherein the mass spectrometry is gas chromatography/mass spectrometry/mass spectrometry (GC/MS/MS).

54. The method of claim 53, wherein said sample undergoes a negative chemical ionization using an ionization gas.

55. The method of claim 53, wherein the parent ion of THCA has an m/z of 620.5.

56. The method of claim 53, wherein the product ion of THCA has an m/z of 492 or 383.

57. The method of claim 53, wherein an internal standard is added to said sample prior to mass spectrometry.

58. The method of claim 57, wherein said internal standard is THCA-d3.

59. The method of claim 58, wherein the parent ion of THCA-d3 has an m/z of 623.5.

60. The method of claim 58, wherein the product ion of THCA-d3 has an m/z of 495 or 386.