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

技术

LC/MS: suitable
solid phase extraction (SPE): suitable

试验参数

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)
mobile phase: [A] 10 mM ammonium acetate; [B] 10 mM ammonium acetate in acetonitrile
flow rate: 0.3 mL/min
column temp.: 35 °C
detector: MS/MS, MRM Transitions, ESI(+) (210.2/180 and 210.2/93.2 m/z)
injection: 20 μL

适用性

application for LC-MS
application for SPE

应用

clinical