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

Sample Preparation Details

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)

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: AB Sciex AB 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

1
JWH-073-Butanoic acid metabolite

2
JWH-018-Pentanoic acid metabolite

3
JWH-073-4-Hydroxybutyl metabolite

4
JWH-073-3-Hydroxybutyl metabolite

5
JWH-018-5-Hydroxypentyl metabolite

6
JWH-018-4-Hydroxypentyl metabolite

7
AM2201-4Hydroxypentyl metabolite

8
AM694
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9
AM2201-d5

10
AM2201

11
JWH-018-Pentanoic acid metabolite

12
JWH018
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