HPLC separation protocol for Pseudomonas aeruginosa pigment Pyocyanin?

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What is the stability of 2 3-dihydroxyquinoxaline in acidic water?

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Robin Joshi

A=ACN

B=0.1%H3PO4,

Start from 90%ACN to 35 and back to 90%

C-18, Gradient elution, flow rate=1ml/min.

You will definitely get your results.

Gud luck!!!

Tomáš Hluska

Hw can you start with 90% AcN?

And flow rate depends on column.

For HPLC, 250x4.6, flow rate is 1ml/min. You have to little optimise on this mobile phase.

But you didn't write that. On any other column it may be different. Even with your column can be the flow set different to affect the resolution and speed.

Anyway, what about the starting conditions?

I know that that column dimension will change its flow rate as in case of UPLC. If you have small or long column the flow rate will be different. Thats why I specially mention the column length and already said that you have to optimise your conditions of HPLC.

Thanks to update me!!!

Zahra Ramezani

How do u