# US 7,465,586 B2 · Oral detection test for cannabinoid use

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 7,465,586 B2
- Document ID: US7465586B2
- Status: Granted
- Kind: B2
- Application number: 10/980,372
- Filed: 2004-11-03
- Published: 2006-05-04
- Granted: 2008-12-16
- Earliest claimed: 2004-11-03
- Assignees: LabOne, Inc.
- Inventors: David Day; David J. Kuntz; Michael S. Feldman
- Web page: https://patentgrep.com/patent/US7465586B2

## CPC

- G01N 33/948: {Sedatives, e.g. cannabinoids, barbiturates (opiates G01N33/9486)}
- Y10S 436/901: Drugs of abuse, e.g. narcotics, amphetamine
- Y10T 436/142222: Hetero-O [e.g., ascorbic acid, etc.]
- Y10T 436/24: Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## 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

### Claim 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-Δ9 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.

### Claim 2 (depends on 1)

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.

### Claim 3 (depends on 2)

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.

### Claim 4 (depends on 3)

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

### Claim 5 (depends on 3)

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

### Claim 6 (depends on 3)

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.

### Claim 7 (depends on 6)

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.

### Claim 8 (depends on 3)

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

### Claim 9 (depends on 8)

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.

### Claim 10 (depends on 1)

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

### Claim 11 (depends on 10)

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

### Claim 12 (depends on 1)

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.

### Claim 13 (depends on 1)

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.

### Claim 14 (depends on 1)

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

### Claim 15 (depends on 1)

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

### Claim 16 (depends on 1)

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.

### Claim 17 (depends on 16)

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

### Claim 18 (depends on 16)

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

### Claim 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-Δ9 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.

### Claim 20 (depends on 19)

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.

### Claim 21 (depends on 20)

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

### Claim 22 (depends on 20)

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

### Claim 23 (depends on 19)

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

### Claim 24 (depends on 19)

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

### Claim 25

A method for determining active intake of marijuana by a subject, comprising:
(i) detecting by an assay an amount of 11-nor-Δ9 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.

### Claim 26 (depends on 25)

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

### Claim 27 (depends on 25)

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.

### Claim 28 (depends on 27)

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.

### Claim 29 (depends on 27)

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

### Claim 30 (depends on 27)

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.

### Claim 31 (depends on 30)

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.

### Claim 32 (depends on 27)

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

### Claim 33 (depends on 22)

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.

### Claim 34 (depends on 25)

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

### Claim 35 (depends on 34)

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

### Claim 36 (depends on 35)

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.

### Claim 37 (depends on 25)

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

### Claim 38

A method for determining active intake of marijuana by a subject, comprising:
(i) detecting by an assay an amount of 11-nor-Δ9 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.

### Claim 39 (depends on 38)

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

### Claim 40 (depends on 38)

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

### Claim 41 (depends on 38)

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

### Claim 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-Δ9 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.

### Claim 43 (depends on 42)

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

### Claim 44 (depends on 42)

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

### Claim 45 (depends on 42)

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

### Claim 46

A method for determining active intake of marijuana by a subject, comprising:
(i) detecting a specified amount of 11-nor-Δ9 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.

### Claim 47 (depends on 46)

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

### Claim 48 (depends on 46)

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

### Claim 49 (depends on 46)

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

### Claim 50 (depends on 46)

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.

### Claim 51 (depends on 50)

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.

### Claim 52 (depends on 46)

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

### Claim 53 (depends on 46)

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

### Claim 54 (depends on 53)

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

### Claim 55 (depends on 53)

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

### Claim 56 (depends on 53)

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

### Claim 57 (depends on 53)

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

### Claim 58 (depends on 57)

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

### Claim 59 (depends on 58)

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

### Claim 60 (depends on 58)

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