HPLC Analysis of β-Blockers and β-Agonists in Urine and Wastewater on a C18 Column after SPE using SupelMIP® SPE-Beta-Receptor
CONDITIONS
sample preparation
SPE (Solid Phase Extraction)
sample/matrix
horse urine or waste water spiked with 10 beta-agonists and beta-blockers at 1 ng/mL
SPE tube/cartridge
SupelMIP SPE - Beta-Receptor, 25 mg/10 mL (LRC) (53223-U)
condition
1 mL acetonitrile; 1 mL DI water
sample addition
1 mL
washing
3 x 1 mL DI water; 1 mL acetonitrile; 1 mL 60% acetonitrile/40% DI water; apply 2 min of full vacuum to dry the tube after each step
elution
2 x 1 mL 1% formic acid in acetonitrile
column
C18, 5 cm x 3 mm I.D., 3 μm particles
eluate post-treatment
evaporate under nitrogen and reconstitute with 150 μL 5% acetonitrile in 10 mM ammonium acetate, pH 4.6 prior to LC-MS-MS analysis
mobile phase
[A] 10 mM ammonium acetate, pH 4.6 (adjusted with acetic acid); and [B] acetonitrile
flow rate
0.5 mL/min
column temp.
ambient
detector
MS/MS, ESI(+)
injection
20 μL
Description
Application
Sample Pre-treatment:
Horse urine was centrifuged at 3000 g for 10 min, diluted 1:1 (v/v) with DI water, adjusted to pH 7. Wastewater was filtered with 1 μm filter paper and adjusted to pH 6-7. Samples were spiked with 10 beta-agonists and beta-blockers at the level of 1 ng/mL.
Legal Information
SupelMIP is a registered trademark of Sigma-Aldrich Co. LLC