A complex mixture of sulphonamide, macrolide, tetracycline and fluoroquinolone antibiotics were separated in one run using mixed-mode chromatography with LC/MS -compatible conditions. All compounds are separated based on reversed-phase and/or ion-exchange mechanism. Method can be used for analysis of various classes of antibiotics and related impurities in different sample matrices (blood, urine, soil, waste water).
Column | Obelisc R, 2.1×150 mm, 5 µm, 100A |
Mobile Phase | Gradient MeCN – 0-25%, 6 min, 25-70% 14 min |
Buffer | Gradient Formic Acid – 0.05%-0.3%, 10 min, 14 min hold |
Flow Rate | 1.0 ml/min |
Detection | UV, 270 nm |
Class of Compounds |
Antibiotic, Drug, Hydrophobic, Ionizable |
Analyzing Compounds | Sulphanilamide, Sulphaguanidine, Oxytetracycline, Marbofloxacin, Sulphadiazine, Chlortetracycline, Nofloxacin, Spiramycin, Cefalonium, Tylosin, Josamycin, Sulphamethoxazole, Dapsone, Sulphaquinoxalline, Flumequine |
SIELC has developed the mixed-mode Obelisc™ columns to be the first commercially available columns with Liquid Separation Cell technology (LiSC™). This cost effective duo can replace multiple HPLC columns such as reversed-phase (RP), AQ-type reversed-phase, polar-embedded group RP columns, normal-phase, cation-exchange, anion- exchange, ion-exclusion, and HILIC (Hydrophilic Interaction Liquid Chromatography) columns. By controlling just three orthogonal method parameters - Buffer concentration, Buffer pH, and Organic Modifier Concentration - users can adjust the column properties with pinpoint precision to separate complex mixtures.
Select optionsPrimesep 100 separates a mixture of sulfonamides (benzenesulfonamide, chlorobenzenesulfonamide, hydroxybenzenesulfonamide) and hydroxysulphaminophenylhydrazine by a mixture of polar and hydrophobic interactions. The stationary phase’s hydrophobic functionality provides a reversed-phase mechanism while the embedded cation-exchange group provides polar interactions. The separation method uses a mobile phase mixture of water, acetonitrile (MeCN, ACN) and trifluoracetic acid (TFA) with UV detection at 210 nm. This method is mass spec (LC/MS) and evaporative light scattering (ELSD) compatible.
The Primesep family of mixed-mode columns offer a wide variety of stationary phases with an unprecedented selectivity in the separation of a broad array of chemical compounds and in multiple applications. Corresponding Primesep guard columns are available with all stationary phases and do not require holders. SIELC offers a method development service which is available for all customers. Ask about our special custom LC-phases tailored for specific separations.
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