PMID- 2393940
OWN - NLM
STAT- MEDLINE
DCOM- 19901009
LR  - 20190824
IS  - 0009-2797 (Print)
IS  - 0009-2797 (Linking)
VI  - 76
IP  - 1
DP  - 1990
TI  - Alternating current voltammetric determination of DNA damage.
PG  - 111-28
AB  - The conditions for alternating current (a.c.) voltammetric DNA determinations 
      have been investigated with respect to its use with alkaline filter elution 
      techniques at low DNA concentrations. In inorganic electrolyte solutions three 
      current peaks can be distinguished: peak I around -1.1 V caused by the 
      reorientation or desorption of DNA segments; peak II around -1.2 V caused by the 
      native DNA (nDNA) form; peak III caused by denatured DNA (dDNA) at -1.4 V. 
      Sonication of nDNA increases the peak current, however not with dDNA. Both dDNA 
      and nDNA give linear peak current increments with DNA increments, their 
      regression lines cutting the concentration axis at the origin. In filter elution 
      techniques organic bases are often used. Adding ethanolamine (EA) elution buffer 
      decreases the peak amplitude of DNA. It turns out that an unknown substance, 
      perhaps a protein or RNA, elutes from the filters and gives rise to a current 
      peak at about -1.3 V. This substance can interfere with the dDNA by competing for 
      electrode surface area, since it diffuses much faster than the large molecules of 
      the DNA. Since however, dDNA has a higher affinity for the electrode surface, 
      after enough time, usually few minutes, the dDNA increasingly displaces the 
      substance and occupies the surface. The same is valid for other organic molecules 
      and thus also for EA. It is therefore remarkable that the unknown substance can 
      be altered by ultrasonication, so that it will no longer interfere with dDNA, in 
      contrast to EA. EA, on the other hand, can be "titrated". When EA is present at 
      short accumulation times it prevents dDNA adsorption. By adding dDNA, the EA can 
      be scavanged and further addition will adsorb and thus increase peak current in 
      proportion to the concentration of the DNA present. The conditions for 
      voltammetric DNA determination have been investigated obeying the recognized 
      interactions. Avoiding organic bases and using inorganic ones would simplify the 
      determination procedure. The reproducibility of the procedure in the range of 
      50-60 ng DNA/ml has been found to be +/- 6%.
FAU - Krznaric, D
AU  - Krznaric D
AD  - Center for Marine Research Zagreb, Ruder Boskovic Institute, Yugoslavia.
FAU - Cosovic, B
AU  - Cosovic B
FAU - Stuber, J
AU  - Stuber J
FAU - Zahn, R K
AU  - Zahn RK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Ireland
TA  - Chem Biol Interact
JT  - Chemico-biological interactions
JID - 0227276
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Animals
MH  - DNA/*analysis
MH  - *DNA Damage
MH  - Electrochemistry
MH  - Hydrogen-Ion Concentration
MH  - Male
MH  - Sea Cucumbers
MH  - Spectrometry, Fluorescence
EDAT- 1990/01/01 00:00
MHDA- 1990/01/01 00:01
CRDT- 1990/01/01 00:00
PHST- 1990/01/01 00:00 [pubmed]
PHST- 1990/01/01 00:01 [medline]
PHST- 1990/01/01 00:00 [entrez]
AID - 0009-2797(90)90038-O [pii]
AID - 10.1016/0009-2797(90)90038-o [doi]
PST - ppublish
SO  - Chem Biol Interact. 1990;76(1):111-28. doi: 10.1016/0009-2797(90)90038-o.