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HPLC Analysis of NNAL in Urine on Ascentis® Express C18 after SPE using SupelMIP® SPE-NNAL

HPLC Analysis of NNAL in Urine on Ascentis® Express C18 after SPE using SupelMIP® SPE-NNAL

Conditions

sample preparation SPE (Solid Phase Extraction)
sample/matrix human urine spiked with 60 or 100 pg/mL NNAL
SPE tube/cartridge SupelMIP SPE - NNAL, 25 mg/10 mL (LRC) (53206-U)
condition 1 mL dichloromethane; 1 mL methanol; 1 mL DI water
sample addition 2 mL (60 pg/mL sample), 5 mL (100 pg/mL sample)
washing 2 x 1 mL DI water, 10 min vacuum; mL toluene; 1 mL toluene:dichloromethane (9:1, v/v); 1 mL toluene:dichloromethane (4:1, v/v), 2 min. vacuum
elution 2 x 1 mL 10% methanol in dichloromethane, apply gentle vacuum between each fraction
eluate post-treatment evaporate under nitrogen and reconstitute in 0.15 - 0.25 mL mobile phase
column Ascentis Express C18, 5 cm x 2.1 mm I.D., 2.7 µm particles (581307-U)
column temp. 35 C
mobile phase [A] 10 mM ammonium acetate; [B] 10 mM ammonium acetate in acetonitrile
flow rate 0.3 ml/min
injection 20 µl
detector MS/MS, MRM Transitions, ESI(+) (210.2/180 and 210.2/93.2 m/z)

Description

Categories Analytical Chromatography, SPE Applications, Clinical, nitrosamines, Pharmaceutical & Clinical applications, Nitrosamines
Featured Industry Clinical
Legal Information Ascentis is a registered trademark of Sigma-Aldrich Co. LLC, SupelMIP is a registered trademark of Sigma-Aldrich Co. LLC
suitability application for LC-MS, application for SPE

Materials

     
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