| 英文摘要 |
This plan established and standardized the MVLN estrogen specific transcription assay according to QC/QA criteria of USEPA method 4425. The evaluated items included establishment of luciferase standard curve, evaluation of solvent selection for sample and used quantity, evaluation of exposure time of sample, establishment of 17b-estradiol (E2) standard curve, and detection sensitivity and precision. The luciferase standard curve gave the equation of y = 2274.6X -1.6944 after linear regression（0.01 ~ 0.16 pg/ml）. The correlation coefficient of the linear regression analysis achieved 0.9985. The precision and detection limit were 1.40 ~ 14.43 %（RSD） and 0.01 pg/ml, respectively. The cytotoxicity of MVLN to DMSO, methylene chloride（MECL）, hexane（HX）, methanol（MEOH）, isooctane（ISOT）and acetone（ACT）decreased in the order of HX＞DMSO＞ACT MEOH＞MECL ISOT. The luciferase induction was increased as the concentration of E2 increased. The standard curve showed sigmoidal pattern having Cmax at 1 nM. Exposure time of MVLN cells to E2 (1~3 days) did not affect significantly their concentration-estrogenicity plots. The precision of E2 in 3 to 6 replicates were 0.18 ~ 15.23 %. The detection limit for E2 exposure time of 1, 2 and 3 days were 1.59 pM, 0.29 pM and 2.54 pM, respectively. The method was further applied to estimate the estrogenicity of 42 plasticizers and interactions between 4 plasticizers. The results revealed that 5 plasticizers as butyl benzyl phthalate（BBP，10 uM）、bis（4-methyl-2-pentyl）phthalate (BMPP，10 uM)、diethoxyethyl phthalate（DEEP，1,10 uM）、hexyl-2-ethylhexyl phthalate (HEHP，10 uM) and monobenzyl phthalate（MBP，10 uM）possessed significant estrogenicity (p<0.01). In case of interaction study, the results implied that BBP, BMPP, DEEP, HEHP and E2 have possibly no interaction each other.
|