Applications
Extraction of COOH-THC from Urine Using Novum SLE & Kinetex 2.6u C8
22786
Separation Mode: Reversed Phase
Reversed Phase
C8
HPLC
Forensics/Toxicology
Toxicology / Drug Screening / TDM
Therapeutic / Clinical

Applications

Extraction of COOH-THC from Urine Using Novum SLE & Kinetex 2.6u C8

Analytes

1
2
Compound Name
Tetrahydrocannabinol-7-Oic Acid
CID
108207
Molecular Formula
C21H28O4
Molecular Weight
344
No. Hydrogen Bond Acceptors
4
No. Hydrogen Bond Donors
2

Conditions

LC Conditions (App ID: 22786)
Column:Kinetex 2.6 µm C8 100 Å, LC Column 50 x 2.1 mm, Ea
Brand Name:Kinetex
Part No:00B-4497-AN
Elution Type:Gradient
Mobile Phase:A: 0.1% formic acid
B: Methanol/acetonitrile(50:50)
Gradient Profile:
Step No.Time(min)%A%B
105050
22.5595
33.5595
455050
Flow Rate:0.5 mL/min
Temperature:25°C
Detection:Tandem Mass Spec (MS-MS) @ 22°C

Sample Preparation Details
Description:Novum SLE, MAX 96-Well Plate, Ea
Part Number:8E-S138-5GA
Pre Treatment Note:Sample Pretreatment Step Enzymatic Hydrolysis: To 300 uL urine add 75 uL of 300 mM ammonium acetate (pH 4.0) and 25 uL of -Glucoronidase solution (100,000 units/mL (www.campbellscience.com, DR2100). Mix and vortex for 30 secs. Incubate at 37- 40°C for 60 minutes. (Gentle shaking during this step is recommended). After incubation, bring samples to room temperature prior to extraction. Extraction Procedure: Sample Loading: Load the sample (~400 uL) from pretreatment step onto the Novum plate and apply a short and gentle pulse of vacuum (~5-10 sec and 5"or less of Hg) until the sample has completely entered in the media. NOTE: Avoid prolonged or excessive vacuum. The sample must not leave the Novum bed. Wait for 5-6 minutes. NOTE: Inadequate or excessive wait period will lead to variable recoveries and poor precision. Elution: Dispense 900 uL of ethyl acetate onto the Novum plate and collect the solvent under force of gravity (approximately 5 min). Repeat with another 900 uL addition of ethyl acetate and collect the solvent under gravity. Apply vacuum at 5" of Hg (or lower) for 20-30 secs to complete the extraction. NOTE: Excessive/Prolong application of the vacuum may force the elution of plasma and cause dirty extraction Dry down: Evaporate extracted samples to complete dryness under slow stream of N2 and room temperature. Reconstitute the dry residue in 200 uL of initial mobile phase containing internal standard
SPE Load:Load 300.00 µL Sample diluted in 75
SPE Elution 1:900 mL Ethyl acetate
SPE Elution 2:900 mL Ethyl acetate
SPE Evaporation:Evaporate to dryness
SPE Reconstitute:200 µL Initial Mobile phase (A:B=50/50)

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