HPLC Analysis of Chloramphenicol in Milk on Ascentis^® C18 after SPE using SupelMIP^® SPE-Chloramphenicol

technique(s)

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

test parameters

sample preparation: SPE (Solid Phase Extraction)
sample/matrix: whole pasteurized milk spiked with chloramphenicol at 15 ng/mL and 38 ng/mL
SPE tube/cartridge: SupelMIP SPE - Chloramphenicol, 25 mg/10mL (LRC) (53210-U)
condition: 1 mL methanol;1 mL DI water
sample addition: 1 mL
washing: 2 x 1 mL MS-grade water, 1 mL 5% acetonitrile in 0.5% acetic acid, 2 x 1 mL MS-grade water, 1 mL 20% acetonitrile in 1% ammonium hydroxide. (dry SPE tubes for 15 min. under gentle vacuum; 3 x 1 mL dichloromethane; dry SPE tubes for 1 min. under gentle vacuum)
elution: elute chloramphenicol with 2 x 1 mL methanol:acetic acid:MS-grade water (89:1:10, v/v/v)
eluate post-treatment: evaporate combined eluate to dryness at 50 °C under nitrogen. reconstitute in 150 mL LC mobile phase prior to LC-MS analysis
column: Ascentis C18, 2.1 mm x 10 cm I.D., 3 μm particles (581301-U)
mobile phase: [A] 100 mM ammonium acetate: [B] MS-grade water: [C] acetonitrile (10:60:30, A:B:C)
flow rate: 0.2 mL/min
column temp.: 35 °C
detector: MS, ESI(-), 320-323 m/z range
injection: 5 μL

suitability

application for LC-MS
application for SPE

application(s)

food and beverages