THC and Metabolites from Whole Blood on Strata-X-Drug B and Kinetex C18 by LC-MS-MS

LC Conditions

Column

Kinetex 2.6 µm C18 100 Å, LC Column 50 x 2.1 mm, Ea

Brand

Kinetex

Part No

00B-4462-AN

Phase Name

C18

Elution Type

Gradient

Mobile Phase

A: 1 mM ammonium formate with 0.1% FA

B: 0.1% FA/methanol:acetonitrile(1:1)

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

50

50

2

3

5

95

3

4.5

5

95

4

4.51

50

50

5

6

50

50

Flow Rate

0.40 mL/min

Temperature

22°C

Detection

Mass Spectrometer (MS) @ (22°C)

Detection Info

Sample Notes

Mass spec conditions: Polarity Analyte Q1 Q3 DP CE (+) THC-OH (*) 331.0 193.3 55 31 (+) THC-OH 331.0 201.2 55 33 (+) THC-OH 331.0 175.3 55 31 (+) THC-OH-d3 334.0 196.3 55 31 (+) THC (*) 315.2 193.1 60 31 (+) THC 315.2 259.1 60 35 (+) THC-d3 318.2 196.1 60 25 (+) THC-COOH (*) 345.2 327.0 55 29 (+) THC-COOH 345.2 299.2 55 49 (+) THC-COOH-d3 348.0 330.1 55 29 CAD: 7 GS1: 60 GS2: 45 TEM: 600 CUR: 25 IS: 5500 * indicates quant ion

Sample Preparation Details

Description

Strata-X-Drug B 33 µm Polymeric Strong Cation, 60 mg / 6 mL, Tubes, 30/Pk

Part No

8B-S128-UCH

Pretreatment Note

Evaporate eluant to dryness under 50 °C nitrogen. Reconstitute in 500uL of initial mobile phase. Extraction was automated and performed using a Gilson GX-274 ASPEC system

Load

Load Sample

Wash 1

2mL of ACN:Water (15:85)

Dry

5 mins at 7-15 psi Nitrogen

Elution 1

2mL of Ethyl acetate:Isopropanol (85:15)

Evaporation

Evaporate to dryness

Reconstitute

500 µL Initial mobile phase (A/B=50/50)