Region

Applications

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

Sample Preparation Details

Part No:8B-S128-UBJ
Solid Phase Extraction (SPE) procedure:Note: The solvent volumes shown below are for a 60 mg bed mass. The solvent volumes will need to be adjusted for a smaller or larger bed mass.
Load: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.

Dry:15 mins under 10-15" of Hg
Final Prep and Analysis: 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
Inject: 10 µL on HPLC / Mass Spectrometer (MS) @ amu (ambient)

Analytes:

Name Spiked
Conc.
Log P pKa % Rec %RSC (n=0)
1 JWH-073-Butanoic acid metabolite 50  60
2 JWH-018-Pentanoic acid metabolite 50  68
3 JWH-073-4-Hydroxybutyl metabolite 50  90
4 JWH-073-3-Hydroxybutyl metabolite 50  90
5 JWH-018-5-Hydroxypentyl metabolite 50  80
6 JWH-018-4-Hydroxypentyl metabolite 50  83
7 AM2201-4Hydroxypentyl metabolite 50  80
8 AM694 50  101
9 AM2201-d5 100 
10 AM2201 50  104
11 JWH-018-Pentanoic acid metabolite 50  89
12 JWH018 50  71

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

9
AM2201-d5

10
AM2201

11
JWH-018-Pentanoic acid metabolite

12
JWH018