I am looking for certain alcoloids in a certain food product. While i was searching related articles and determination methods, i recognised some of them indicated pH for mobile phase. However, not all articles specify pH conditions of mobile phase. Should i determine pH of mobile phase for each modification about alcoloids? What is its tricky point?
In General, To optimize a pH of Mobile phase is most important part in HPLC method development. The attach file might be useful to you. So, Please find attachment.
Dear Elif Öztürk,
There are seven parts that make HPLC, are connected together and are equally important such as column, mobile phase, pump, detector... etc. Before to come to your question one of those important parts is the mobile phase, that is used to optimize the separation of interested compounds nicely (good resolution) and with enough distance from each other or adding some extra material such as TFA to eliminate tailing in the peak or control the pH to optimize best separation. Either you use isocratic ( a mobile phase with one bottle with stable concentration over the separation process. Or using gradient elution (two bottles or more that their concentration is always changing over the separation process. Now your question ''Should i determine pH of mobile phase for each modification about alkaloids? What is its tricky point? No you may not need to control the Ph so how do I know? from literature see the suggested mobile phases, then take a decision according to the above conditions: does this mobile gives good resolution, separated peaks from each other nicely...etc so start to develop your method try several mobile phases whether modified pH or not or even to change your wavelength, your column. To develop a method you need 2-3 months but that will give much experience when work with HPLC.
Best regards
Aly R Abdel-Moemin
Dear research scientists,
While we are preparing samples for HPLC, should we use mobile phase or can we use distilated water? Is it obligatory to use mobile phase while preparing samples?
For example asetonitrile:sodium aceate is used for mobile phase and then prepare samples with distilled water. Or this treatment is wrong ? Should i prepare my samples with mobile phase,too (Note: i use C18 colon. It is apolar colon type)
I couldn't reach file.
Thank you for your attention.
Yours Sincerely,
Elif ÖZTÜRK
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