Skip to Content
MilliporeSigma

HPLC Analysis of Tetracycline Antibiotics on Ascentis® Express C18

HPLC Analysis of Tetracycline Antibiotics on Ascentis® Express C18 application for HPLC

Materials

analytical column

Product No.
Description
Pricing

Ascentis® Express C18, 2.7 μm HPLC Column

2.7 μm particle size, L × I.D. 15 cm × 3 mm

also commonly purchased with this product

Product No.
Description
Pricing

Vials, screw top, silane-treated

volume 2 mL, amber glass vial, O.D. × H × I.D. 12 mm × 32 mm × 4.6 mm

guard column

Product No.
Description
Pricing

Ascentis® Express C18, 2.7 Micron Guard Cartridge

2.7 μm particle size, L × I.D. 5 mm × 3 mm, pkg of 3 ea

mobile phase component

Product No.
Description
Pricing

Oxalic acid dihydrate

ACS reagent, ≥99%

standard

Product No.
Description
Pricing

Chlortetracycline hydrochloride

VETRANAL®, analytical standard

Oxytetracycline hydrochloride

VETRANAL®, analytical standard

4-Epitetracycline hydrochloride

VETRANAL®, analytical standard

Doxycycline hyclate

VETRANAL®, analytical standard

Tetracycline hydrochloride

VETRANAL®, analytical standard

CONDITIONS

column

Ascentis Express C18, 15 cm x 3.0 mm I.D., 2.7 μm particles (53816-U)

mobile phase

[A] 10 mM oxalic acid in water; [B] 5 mM oxalic acid in methanol:acetonitrile (1:1)

gradient

15% B for 1 min; to 20% B in 0.1 min, held at 20% for 4 min; to 80% B in 5 min; to 15% B in 0.1 min, held at 15% B for 3 min

flow rate

0.5 mL/min

pressure

3597 psi (248 bar)

column temp.

30 °C

detector

UV, 355 nm

injection

50 μL

sample

100 ng/mL of each compound in acetontrile:water:formic acid (5:94.9:0.1)

Description

Analysis Note

Separation of tetracycline antibiotics from their epi-metabolites was achieved on an Ascentis® Express C18 HPLC column. Oxalic acid was added to the mobile phase to prevent the loss of these metal-binding compounds in the HPLC system.

Legal Information

Ascentis is a registered trademark of Merck KGaA, Darmstadt, Germany