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HPLC polyphenols


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15 replies to this topic

#1 hidayahcellculture

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Posted 07 October 2013 - 09:18 PM

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



#2 bob1

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Posted 08 October 2013 - 01:00 PM

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.



#3 mdfenko

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Posted 09 October 2013 - 03:46 AM

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).


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#4 hidayahcellculture

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Posted 09 October 2013 - 11:39 PM

means i need to do gc ms?



#5 mdfenko

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Posted 10 October 2013 - 04:01 AM

lcms would probably suit your needs better than gcms.


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#6 hidayahcellculture

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Posted 13 October 2013 - 11:25 PM

lcms would probably suit your needs better than gcms.

do you have protocol for icms?



#7 bob1

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Posted 13 October 2013 - 11:34 PM

LCMS - liquid chromatography MS...



#8 hidayahcellculture

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Posted 19 October 2013 - 08:49 PM

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?



#9 bob1

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Posted 21 October 2013 - 12:06 AM

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.

#10 ckzimase

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Posted 09 November 2013 - 01:44 AM

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.



#11 hidayahcellculture

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Posted 23 November 2013 - 09:15 PM

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.

 do you have any example procedure for LCMS?



#12 hidayahcellculture

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Posted 25 December 2013 - 10:19 PM

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.

if anybody could identify them? i will give the result for the peaks obtained....



#13 mdfenko

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Posted 26 December 2013 - 04:13 AM

giving us the results will be meaningless without reference standards with which to compare (and too tedious to compare without proper software).

 

which hplc system are you using?

 

did it come with software? if so, is it just for control or also for analysis?


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#14 ckzimase

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Posted 26 December 2013 - 11:08 PM

can u give me your email add so i can send it to you



#15 hidayahcellculture

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Posted 27 December 2013 - 09:13 AM

can u give me your email add so i can send it to you

my email: hhhhhhhhhh586@yahoo.com






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