Chromatography Forum

Iam using a method for Tacrolimus HPLC analysis for the separation of Tautomers.
MP: 600:400:1 Acetonitrile : h2O : Phosphoric acid
210 nm, 50 deg column temperature.
Sample prepared on a brij diluent.
But iam unable to get any peaks in HPLC.
Can anybody guide on this issue.

Dear Jijualex,


A) Did you check pertinent literature about this analysis?
See this good article "Determination of the immunosuppressive drug tacrolimus in its dosage forms by high-performance liquid chromatography"
Y. Namiki, A. Fujiwara, N. Kihara, S. Koda, K. Hane and T. Yasuda - Chromatographia, Volume 40, Numbers 5-6, 253-258, DOI: 10.1007/BF02290354 http://www.springerlink.com/content/t44v44v032382rm7/

B) I already worked with FK506 but I decided to move to LC-MS for two pivotal reasons:

i) the better analytical responses for this analyte occours at worst region of Deuterium lamps, ie, around 204nm to 222nm;

ii) BUT the UV absortivity of Tacrolimus is very (much, much, much!) low even at 204nm to 222nm.

If you had used diluted solutions detection problems had followed you.

Consider it and your real possibilities.

C) By other way, are you sure your HPLC system has working properly?

D) Be free for e-mail me with additional details about your FK506 project, since I may help you.


Hoping for the better,

Carlos de Souza Teixeira
teixeiracs@yahoo.com

Thanks for your valuable advice.

I used the method from the inventor company publication

Quantitative determination of tautomeric FK506 by
reversed-phase liquid chromatography
Takeshi Akashi*, Tomoko Nefuji, Masaru Yoshida, Jyunji Hosoda
Fermentation Development Laboratories, Fujisawa Pharmaceutical Co., Ltd., Nakagawara, Shinkawa, Nishikasugai-Gun, Aichi 452, Japan

My HPLC is working properly. Iam able to get Tacrolimus peaks on HPLC with 80% MEOH in water but all tautomers together.

Im working on API of this product so i have to analyse for all tautomers in particular.

The literature you mentioned will try.


Thank you so much for now.....................

Hi you have to dissolve FK506 in water and wait for 12 hours to obtain the tautomers. I worked with UV detection, and it was very usefull in the dissolution test.

Good luck