Neomycin presents a number of challenges to routine Chromatographic Analysis. It lacks Chromophores and therefore is difficult to detect using conventional HPLC techniques and retention in traditional Reversed Phase mode may not be viable due to its high polarity.  However, use of the Cogent Diamond Hydride Column in conjunction with a Mass Spec helps circumvent these issues. The presented data illustrates how the Compound can be both readily retained, with good run-to-run precision, and adequately detected using Mass Spectrometry.

Peak:
Neomycin 615.3196 n/z (M+H)+

Method Conditions:
Column: Cogent Diamond Hydride™, 4µm, 100Å
Catalog No.: 70000-05P-2 
Dimensions: 2.1 x 50 mm
Solvents: DI H2O/ 0.1% Formic Acid (v/v)
Gradient:

Time (minutes) %B
0 90
0.5 90
4 10
5 10
6 90
10 90

Injection Volume: 5µL
Flow Rate: 0.3 mL/minute
Detection: ESI – POS – Agilent 6210 MSD TOF Mass Spectrometer
Sample Preparation: 0.1 mg/mL Neomycin Sulfate Reference Standard Solution in Solvent A Diluent

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