Posted: Fri Feb 11, 2005 6:32 pm
by AA
Chris,
Post up the conditions, I would love to try them. I have little doubt that freshly distilled THF would be better than stabilized, but as you point out, most (including me) would be unwilling to do that.
AA
Improve Conditions for Aminoglycoside Analysis
Posted: Sat Feb 12, 2005 10:58 pm
by Chris Pohl
AA,
I'm not sure which aminoglycosides you're interested in but I've taken these conditions from a recently developed method for Sisomicin and Netilmicin. The column used is the Acclaim PA (from Dionex) 2.1 x 150 mm. Temperature: 40°C. Eluent: 800mM HAc/NaAc (pH 5), 1mM sodium octanesulfonate, 1% acetonitrile (12.96 mL of 100% HAc, 47.12g NaAc, 10 mL acetonitrile, 10 mL 0.1M OSA Na to a final volume of 1.0 L). Flow rate: 0.25 mL per minute. IPAD detection at 0.05 volts versus a silver / silver chloride reference electrode (waveform below). 3 mm gold working electrode. Gasket thickness: 5 mil (0.127 mm). Post column addition of 0.5 molar sodium hydroxide at 0.25 mL per minute.
Waveform (time in seconds):
Time 0.00, 0.05 Volts
Time 0.20, 0.05 Volts (start integration)
Time 0.40, 0.05 Volts (stop integration)
Time 0.41, -2.0 Volts
Time 0.42, -2.0 Volts
Time 0.43, +0.6 Volts
Time 0.44, -0.1 Volts
Time 0.50, -0.1 Volts