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
Details
LC Conditions (App ID: 22786)
Column:
Brand Name:
Part No:
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:
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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