I wanted to estimate the oxidative DNA damage through 8-OHdG and 2'dG estimation through HPLC. Now, the papers I've read mention the use of electrochemical detectors for HPLC, can we perform a similar analysis using PDA detector in HPLC?
i don't think so.
8-oxodGuo basal levels (human cell lines /or human leukocytes) generally not exceed ~ 1500 fmol/100 µg hydrolyzed DNA. The PDA detector quantification limit reaches ~ 100 pmol (on-column). The best option is to perform the analysis through a mass spectrometry system.
regards
I agree with Tiago, unless you have very large amounts you will not get a detector response with the PDA. Also the chemistry of the detectors differ, the PDA uses absorption to detect compounds while the ECD uses oxidation/reduction potentials to detect compounds. Thus, the PDA won't differentiate between normal and damaged DNA products unless you have a special LC column to separate them. If you don't have access to an ECD your next best detector would be the MS or even MS/MS.
A good detector is the Mass Spectrometer. We have performed some applications on this compound. The references are:
1) A serially coupled stationary phase method for the determination of urinary 8-oxo-7,8-dihydro-2'-deoxyguanosine by liquid chromatography ion trap tandem mass spectrometry.
Rota C, Cristoni S, Trenti T, Cariani E.
Redox Biol. 2013 Oct 16;1:492-7. doi: 10.1016/j.redox.2013.10.001. eCollection 2013.
PMID:
24251117
2) J Pharm Biomed Anal. 2003 Aug 8;32(4-5):657-61.
LC-APCI-MS/MS analysis of urinary 8-hydroxy-2'-deoxyguanosine.
Pietta PG1, Simonetti P, Gardana C, Cristoni S, Bramati L, Mauri PL.
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