HPLC-UV Analysis of Penicillin G in Milk Using an Ascentis® Express C18
Materiales
related product
used together
Sistema de purificación de agua ultrapura Milli- Q® IQ 7000
output: type 1 water (18.2 MΩ·cm), the most advanced Milli-Q® ultrapure (Type 1) water purification system that is intelligent, intuitive, and green.CONDITIONS
column
Ascentis® Express C18, 2.7µm, 100x2.1mm (53823-U)
mobile phase
[A] 20 mM Potassium dihydrogen phosphate in water; [B] Methanol; A/B 60/40 (V/v)
gradient
Isocratic
flow rate
246 µL/min
pressure
195 bar (2828 psi)
column temp.
30 °C
detector
PDA, UV at 280 nm (flow cell: 11 µL/10 mm)
injection
1 µL
sample/matrix
Spiked milk sample: 2 mL of the milk sample was pipetted into a 15 mL polypropylene centrifuge tube, and 20 μL of penicillin G sodium (5 mg/mL) was added to give a final concentration of 50 μg/mL. The mixture was vortexed and incubated at 25 °C for 3 hours under mild agitation. 8 mL of acetonitrile was added to allow sufficient precipitation of proteins. After centrifugation (2,600 rpm, 30 min, 25 °C) and filtration (0.22 μm), an aliquot of the resultant solution was injected into the HPLC.
Descripción
General description
Penicillin G sodium (Benzylpenicillin) is a beta-lactam narrow-spectrum antibiotic used to treat bacterial infections such as pneumonia, syphilis, and other conditions. It is also utilized in the treatment of animals raised for meat or milk production. Discovered by Alexander Fleming in 1929, benzylpenicillin is listed as an essential medicine by the World Health Organization (WHO). Here, an analytical procedure was performed using a 100x2.1 mm C18 Fused-Core® particle column with a UHPLC- system and UV detection at 280 nm.
Analysis Note
Penicillin G eluted in less than four minutes, with a good peak shape and well resolved from the matrix components, on a 100x2.1mm Ascentis® Express C18 (2.7 µm particle) column. For the HPLC method, a detection limit (LOD) of 0.01 µg/mL and a limit of quantitation (LOQ) of 0.03 µg/mL can be achieved at 280 nm.
Other Notes
APP_392
Legal Information
Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany
Fused-Core is a registered trademark of Advanced Materials Technology, Inc.