HPLC polyphenols - (Oct/08/2013 )

Hi everybody, Hi all botanists

need your help regarding HPLC

i use hplc to detect polyphenols present in my plant extract

the compounds detect are many and the chromatogram peaks are clear

is there any library regarding hplc that i can refer in term of retention time to ensure what compounds are present in my plant extract

HPLC doesn't work like that, there could be many different compounds present that could run at the same rate through a column.  Once you have identified your compounds, you can be reasonably sure that the peaks will be consistent, and so use the presence of a peak with the right retention time to identify your compounds. 

To identify the compounds you would probably need to do some sort of mass-spectroscopy.

if you can confirm that your peaks are pure then you can compare them to standards (purified samples of all the compounds that you want to identify, run separately and in combination) run with the same conditions (and, possibly, normalized with an internal standard).

bob1's suggestion to use ms is spot on, especially if you want to confirm identity and/or purity of the peak (you should still run standards).

means i need to do gc ms?

lcms would probably suit your needs better than gcms.

mdfenko on Thu Oct 10 12:01:57 2013 said:

lcms would probably suit your needs better than gcms.

do you have protocol for icms?

LCMS - liquid chromatography MS...

bob1 on Mon Oct 14 07:34:15 2013 said:

LCMS - liquid chromatography M

 

 

for hplc analysis that were done, there were too many peaks appeared and i think  it is useless if i just ignore them and do again with LCMS.

what is your opinion?

You may not need to discard those results; in a new run you should be able to identify the peaks by LCMS and then match those back to your original data.

yes, you must do MS. LCMS is better. GCMS is usually for volatile compound. After HPLC, u can run LCMS. or HPLC-MS. OR if u have standard, u can make standard curve of polyphenols in HPLC and compare.