For identifying components, you should have standards (DRUG). Inject commercial standard and find retention time of compound using same method. Find compare retention time of standard with chromatogram obtained for your crude sample. If proper standars are not available, then better collect individual fractions and do Mass spectrometry.
I believe you know the nature of the sample before injecting. I am pointing towards the expected components on the basis of chemical composition. If you have standards, its well and good, otherwise you can search the literature about your plant and find the Rt for the expected compounds and compare it finally with your HPLC chromatogram to see if it is the one present in your sample too.
If you are blank regarding the constituents, then first you should run an IR and NMR and get the nature of chemical category present in your sample. Then it will be easier for you to proceed.
Gud Luck
As Kalivarathan Divakar said, u need to run the standard phyto molecules using same experimental condition and compare both chromatogram of standard molecules and crude drug for retention time.
lots of time retention time varies in same protocol of literature using HPLC, so it is not exact method to identify your compound. if the standards are not available then u have two ways, first is to search synthetic compound (reference compound) which have almost same properties of your compound and by spiking it in your sample and after analysis if it is increases then u can say that your compounds is present.
Second is just go for MS and identify your compound, so u can say that your target compound is present or not. but it will use only for qualitative analysis, for quantitative analysis u need standards or other reference standards.
hi Fiaz,
Reviewer told u about the Chromatogram of your compounds compare with the standards. hv u give any details about quantification? if you gave, then you have to submit the chromatograms of standards with your samples, otherwise you can not say that the present compounds are your interested compounds using HPLC system with UV detector or you can give the detail of MS if u used.
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