[The formation of BPDE-DNA adduct in rat and its determination by high performance liquid chromatography].
OBJECTIVE
To study the benzo (a) pyrene (BaP) induced DNA adduct in rat and establish a method to measure the DNA adduct in blood by high performance liquid chromatography (HPLC).


METHODS
The SD rats were treated with 100 mg/kg of BaP-DMSO (cosolvent: 1% sodium carboxymethyl cellulose) once by i.p., and the blood of femoral vein was collected 5 hours later. The blood DNA was extracted by kit and confirmed by agarose gel electrophoresis. After extraction, the DNA adducts were hydrolyzed in 0.1 nmol/L HCl at 90 degrees C for 4 hours. The acid-hydrolysis products (BP-tetrols) of DNA adducts were extracted by ethyl acetate and measured by HPLC, and finally confirmed by HPLC-MS.


RESULTS
In chromatogram there were new peaks to occur for rats treated by BaP, with compared to control. By HPLC-MS, one of the new peaks was confirmed to be BP-tetrol.


CONCLUSIONS
The rats ingesting BaP in the way described in this experiment can form DNA adducts, and the adducts in blood can be detected by HPLC.
Yang
Liu-Hua
LH
Department of Sanitary Technology, West China School of Public Health, Sichuan University, Chengdu 610041, China.
Liu
Qu
Q
Sun
Cheng-Jun
CJ
Guo
Shu-Yan
SY
Lin
Shao-Jie
SJ
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000737:chemistry; D000818:Animals; D001564:Benzo(a)pyrene / Q000737:chemistry / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004247:DNA / Q000097:blood / Q000737:chemistry; D018736:DNA Adducts / Q000097:blood* / Q000737:chemistry; D004249:DNA Damage; D004587:Electrophoresis, Agar Gel; D005260:Female; D008297:Male; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley
D018736:DNA Adducts; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; D001564:Benzo(a)pyrene; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D004247:DNA