i have extracted some bacterial peptides and i wanted to purify them using HPLC but i dont know how can i purify these peptides. i need suggestions for doing this work. what program should i use in HPLC so that i can detect all peptides e.g. surfactin, fengycin, bacillomycin etc at the same time using single program.
First of all hplc is not a magic black box. Second of all you need to provide details for people to help you. What columns do you have? where are these lipopeptides dissolved in? What other molecules are in solution? sizes of the peptides you want to separate? Solubility of these peptides in water and in organic solvents?
Perhaps these papers might give you some hints:
Article Purification of Bioactive Lipopeptides Produced by Bacillus ...
http://www.jmb.or.kr/submission/Journal/020/JMB020-01-19_FDOC_2.pdf
https://pdfs.semanticscholar.org/28da/26efca9c0fc695ae285a103fe2b3b34ee6f3.pdf
https://www.intechopen.com/download/pdf/51664
I would like to use high-recovery RP-HPLC method. Small column size (less than 50 mm length with 10 micro-meter particle diameter) is good and sufficient for gradient elution. Acidic phosphate buffer system (pH 2.0) is suitable to silica gel. Good silica gel should be used; i.e., Nucleosil of Macherey & Nagel Co., Düren, Germany, and Develosil of Nomura Chemical, Seto-city, Aichi, Japan are reliable and robust to be used (please see file; Lysozyme by RP-HPLC).
Protein separation should be done at low temperature in order to prevent denaturation, however small proteins or peptides (less than relative molecular mass Mr 1,500) can be separated at higher temperatures with lower column-inlet pressures.
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