By RVJAYANTHI on Monday, December 11, 2000 - 09:40 pm:
We have extracted Ergosterol from musty Agri product and HPLC'd on RP18 reverse column with Methanol mobile phase.The chromatogram is very crowded.
How to make peaks sharp and get a clean chramatogram.
Can someone give me a good method for separation and identificaton of Ergosterol?
By Anonymous on Tuesday, December 12, 2000 - 05:32 am:
Rapid determination of ergosterol in yeast cells
Arnezeder, C. et al.
Anal. Chim. Acta, 1989, 225, Page 129
Analysis of free and esterified ergosterol in tomato products
de Sio, F.; Laratta, B.; Giovane, A.; Quagliuolo, L.; Castaldo, D.; Servillo, L.
J. Agric. Food Chem., 2000, 48, Page 780-784
By Anonymous on Tuesday, December 12, 2000 - 11:05 pm:
Schwadorf, K., Müller, H.M., 1989: Determination of Ergosterol in Cereals, Feed Components, and Mixed Feed by liquid Chromatography. J. Assoc. Off. Anal. Chem. 72, 457-462.
We got suitable results, regarding gras-extracts with Methanol(98%) + Water(2%), using different columns (Prodigy II ODS3 5µm 250x4, Inertsil ODS3 250x4 5µm, even a shorter length does well)
By bill tindall on Wednesday, December 13, 2000 - 08:00 pm:
Gardner, etc,..Ergosterol Determination in activated sludge and its application as a biochemical maker for monitoring fungal biomass. Microbiol. Methods, vol17 49-60 (1993) paper covers sample preparation, novel fast hydrolysis of ergosterol esters followed by lc determination. Confirmation by MS. I can't imagine a nastier sample than waste water treatment sludge and the chromatograms were clear in elution window for analyte.
By bill tindall on Wednesday, December 13, 2000 - 08:09 pm:
P.S. We found the ergosterol esters resistant to hydrolysis in our samples. Hence, a sledge hammer of a soponifaction was called for. see Anal. Chem 63,1251 (1991) for saponification reagent details
By H W Mueller on Monday, December 18, 2000 - 08:09 am:
As an advisor to one of the "anonymous" it seems appropriate to expound on results. This being a "real world problem" the chromatograms optained here were also quite crowded, but the ergosterol was overtowering its neighborhood in many samples to an extend which left no doubt regarding the presence of ergosterol. On the other hand, such methods, with one column in one system, appear not to be cabable to prove the absence of ergosterol.
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