TCA cycle intermediates!

[MS-NMR]

Hello!

I`m in the LC-MS field new. To start I must develope a method to seperation and detection of TCA cycle intermediates such as citrate, fumarate, malate and TCA cycle-related amino acids such as glutamine using LC-MS. Could someone help me how can i start. I mean which column? which mobile phase?
Thank you very much

[sassman]

Look into Hydrophilic interaction liquid chromatography (HILIC). I know there are a few posts on this board about companies that sell columns etc. Use this with ESI-MS. It sounds like you mostly want to quantify, so Triple Quad (ABI or Waters?) If you also want to look at unknowns, bio, etc, then consider Q-TOF or Orbitrap. Maybe even a conductivity detector would do it?

[kerri]

Did a quick search on Science Citation and found this:

Qu, H; Sonnewald, U. “Glutamate is preferred over glutamine for intermediary metabolism in cultured cerebellar neurons.â€

[RB]

Hello
What media are you trying to analyze? Cell extracts? I would suggest HILIC but it also depends on your samples. I have some experience the analysis of TCA metabolites, so feel free to call or send me an email if you need more help. As a start I can recommend this article ‘J. Agric. Food Chem, 2007, 55, 551-560’.
Best regards,
/Rune

[MS-NMR]

Look into Hydrophilic interaction liquid chromatography (HILIC). I know there are a few posts on this board about companies that sell columns etc. Use this with ESI-MS. It sounds like you mostly want to quantify, so Triple Quad (ABI or Waters?) If you also want to look at unknowns, bio, etc, then consider Q-TOF or Orbitrap. Maybe even a conductivity detector would do it?

Dear Sassman!

Thank you very much!

[MS-NMR]

[quote="kerri"]Did a quick search on Science Citation and found this:

Qu, H; Sonnewald, U. “Glutamate is preferred over glutamine for intermediary metabolism in cultured cerebellar neurons.â€

[MS-NMR]

Hello
What media are you trying to analyze? Cell extracts? I would suggest HILIC but it also depends on your samples. I have some experience the analysis of TCA metabolites, so feel free to call or send me an email if you need more help. As a start I can recommend this article ‘J. Agric. Food Chem, 2007, 55, 551-560’.
Best regards,
/Rune

Dear RB!

Thank you for your reply! I'll be very very happy if you can help me.
I'm going to use cell extracts but I'm not sure whether extraction with PCA (perchloric acid) is good or I must try another extraction method.
My email is nmr13c@gmail.com I'll be very happy if I hear from you.
Thank you