Buprenorphine & Norbuprenorphine

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Matrix

Whole blood

Analytes

Buprenorphine, Norbuprenorphine

SPE Cartridge

SiliaPrepX SCX
30 mg / 3 mL
SPE-P0005-03AA

Sample Pretreatment before Loading

Dilute 0.5 mL of blood with 3 mL of Phosphate buffer (0.1 M, pH 6.0). Vortex, centrifugate and discard cellular debris.

Conditionning

3 mL of Methanol

Equilibration

3 mL of Water

Loading

Pretreated sample

Washing

2 x 2 mL of 2 % Formic Acid
3 mL of Methanol / 2 % Formic Acid (70:30)
Dry the cartridge

Elution

2 x 1.5 mL of 5 % Ammonium Hydroxide in Ethyl Acetate / Isopropanol (80:20)

Further Treatment

Evaporation under Nitrogen, reconstitution with 0.1 % Formic Acid in Methanol / Water (15:85) and quantification by LC/MS/MS.