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Extraction of Synthetic Cannabinoids in Human Urine Using Strata-X-Drug B

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Sample Preparation Details

Part Number :8B-S128-UBJ

Description :Strata-X-Drug B 33 µm Polymeric Strong Cation, 60 mg / 3 mL, Tubes, 50/Pk

Load :Load 1.00 mL Sample diluted in 100 mM sodium acetate buffer, pH 5.0

Wash 1 :2 mL 100 mM sodium acetate buffer, pH 5.0 at 1 mL/min

Wash 2 :2 mL Acetonitrile: 100 mM sodium acetate buffer, pH 5.0 (30/70) at 1 mL/min

Dry :15 mins under 10-15 of Hg

Elution 1 :2 mL Ethyl acetate:Isopropanol (85/15) at 1 mL/min

Evaporate: :Evaporate to dryness

Reconstitute :0.50 mL 500 uL of initial HPLC mobile phase (A/B=55/45)

Notes :Extraction Protocol (No need for conditioning or equilibration of the cartridge) Load: Pre-treated samples from hydrolysis (as below) Sample Hydrolysis: Combine 1 mL Human Urine sample (spiked with analytes at 50 ng/mL), 2 mL of 100 mM sodium acetate buffer, pH 5.0, 25 uL -D-Glucoronidase (Patella Vulgata from Sigma, 100KU). Vortex 10-15 secs, followed by incubation for 2 hours in a shaker at 55o C to complete hydrolysis of the glucuronides. Wash 1: 2 mL 100 mM sodium acetate buffer, pH 5.0 Wash 2: 2 mL Acetonitrile: 100 mM sodium acetate buffer, pH 5.0 (30/70) Dry: 15 mins under full vacuum Elute: 2 ml Ethyl acetate:Isopropanol (85/15) Blow Down: To dryness under nitrogen at 40-45o C Reconstitute: With 500 uL of initial HPLC mobile phase (A/B=55/45) spiked with Internal standard at 100ng/mL

Application Detail (App ID: 20925)

Column: Kinetex® 2.6 µm C18 100 Å, LC Column 150 x 3 mm, Ea

Part No:00F-4462-Y0

Dimensions:150 x 3 mm ID

Elution Type:Gradient

Elution A:10mM ammonium formate

Elution B:Acetonitrile

Gradient Profile:

Step No.	Time (min)	Pct A	Pct B
1	0	55	45
2	7	50	50
3	7.01	5	95
4	10	5	95

Flow Rate:0.6 mL/min

Col. Temp: ambient

Detection: Mass Spectrometer (MS) @ amu (22°C)

Detector Info: SCIEX SCIEX API 4000™

Note:Polarity: Positive

CAD: 7.00
CUR: 20.00
GS1: 50.00
GS2: 50.00
IS: 5500.00
TEM: 550.00
ihe: ON
EP 10.00

Analyte Details

Analyte Detail not available

JWH-073-Butanoic acid metabolite

JWH-018-Pentanoic acid metabolite

JWH-073-4-Hydroxybutyl metabolite

JWH-073-3-Hydroxybutyl metabolite

JWH-018-5-Hydroxypentyl metabolite

JWH-018-4-Hydroxypentyl metabolite

AM2201-4Hydroxypentyl metabolite

AM694

AM2201-d5

AM2201

JWH-018-Pentanoic acid metabolite

JWH018