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

LC Conditions

Column

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

Brand

Kinetex

Part No

00F-4462-Y0

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 10mM ammonium formate

B: Acetonitrile

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

55

45

2

7

50

50

3

7.01

5

95

4

10

5

95

Flow Rate

0.60 mL/min

Temperature

22°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Notes

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

Sample Preparation Details

Description

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

Part No

8B-S128-UBJ

Pretreatment Note

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

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

Evaporation

Evaporate to dryness

Reconstitute

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