Aminopyrenes

Fomblin H-Vac 1-aminopyrene stabilization and enhancement 50% in 1,1,2-trichloro-trifluoroethane [238]... 

Amino-3-nitrotoluene 1 b 415,419 Amino phases la 3 Aminophenazone lb312,314,354 2-Aminophenol lb 309, 310, 381 4-Aminophenol lb 309, 310, 381 Aminophenols lb 309,381,383,401 9-(p-Aminophenoxy)acridine lb 145 1-Aminopyrene la 61 4-Amino salicylic acid lb 309,310 Amino sugars lb 47,232-235,354 Aminotrimethylenephosphonic acid la 172... 

Guttman, A. and Pritchett, T., Capillary gel electrophoresis separation of high-mannose type oligosaccharides derivatized by l-aminopyrene-3,6,8-trisul-fonic acid, Electrophoresis, 16, 1906, 1995. 

Compared to the extensive data that have been obtained on the mutagenicity of nitro PAHs in S. typhimurium, relatively little is known about the metabolism of these compounds in this organism. Messier et al. (67) reported that incubation of 1-nitropyrene with S. typhimurium TA98 yielded 1-aminopyrene and 1-acetylaminopyrene as major and minor metabolites, respectively. The reduction of 1-nitropyrene was slow and was accompanied by a slow formation of DNA adducts. When incubations were conducted with the nitroreductase-deficient strain, TA100 F50, both the extent of 1-amino-pyrene formation and DNA binding decreased. Howard ej al. (71,115) also found reduction of 1-nitropyrene to 1-aminopyrene in strains TA98, TA1538 and ATCC 14028. 

Although 1-aminopyrene is a reduced metabolite of 1-nitropyrene, this arylamine will not covalently bind to DNA in vitro (72). In contrast, when incubations were conducted with the intermediate reduction product, N-hydroxy-l-aminopyrene, extensive covalent binding to DNA was detected (72). This observation is consistent with the previous report that several N-hydroxy arylamines formed DNA adducts and induced mutations in S. typhimurium (116), and suggests that, at least for 1-nitropyrene, reduction to N-hydroxy-l-aminopyrene is a critical step in mutation induction. 

Other bacteria. Intestinal bacteria may play a critical role in the metabolic activation of certain nitroaromatic compounds in animals (119) and several reports have appeared on the metabolism of nitro PAHs by rat and human intestinal contents and microflora (120-123). Kinouchi et al. (120) found that 1-nitropyrene was reduced to 1-aminopyrene when incubated with human feces or anaerobic bacteria. More recently, Kinouchi and Ohnishi (121) isolated four nitroreductases from one of these anaerobic bacteria (Bacteroides fragilis). Each nitroreductase was capable of converting 1-nitropyrene into 1-aminopyrene, and one form catalyzed the formation of a reactive intermediate capable of binding DNA. Howard ej al. (116) confirmed the reduction of 1-nitropyrene to 1-aminopyrene by both mixed and purified cultures of intestinal bacteria. Two additional metabolites were also detected, one of which appeared to be 1-hydroxypyrene. Recently, similar experiments have demonstrated the rapid reduction of 6-nitro-BaP to 6-amino-BaP (123). 

Nitropyrene. 1-Nitropyrene is the principal nitro PAH found in diesel exhaust (40) and, therefore, has been the subject of intense study. Nachtman and Wei (133) found that under anaerobic conditions, 1-nitropyrene was reduced by hepatic S9, cytosol or microsomes to principally 1-aminopyrene. Only limited reduction occurred in the absence of cofactors, while maximum metabolism was observed in the presence of both FMN and NADPH. Although the microsomal fraction had the greatest specific activity toward 1-nitropyrene metabolism, the cytosol had 30 times the total activity. 

Saito et al. (134) found that the cytosolic nitroreductase activity was due to DT-diaphorase, aldehyde oxidase, xanthine oxidase plus other unidentified nitroreductases. As anticipated, the microsomal reduction of 1-nitropyrene was inhibited by 0 and stimulated by FMN which was attributed to this cofactor acting as an electron shuttle between NADPH-cytochrome P-450 reductase and cytochrome P-450. Carbon monoxide and type II cytochrome P-450 inhibitors decreased the rate of nitroreduction which was consistent with the involvement of cytochrome P-450. Induction of cytochromes P-450 increased rates of 1-aminopyrene formation and nitroreduction was demonstrated in a reconstituted cytochrome P-450 system, with isozyme P-448-IId catalyzing the reduction most efficiently. 

In the PO-CL system, the compounds showing native fluorescence or that fluoresce after chemical derivatization can be detected. As examples of the PO-CL detection of native fluorescence compounds, dipyridamole and benzydamine in rat plasma [57] and fluphenazine [58] have been reported in the former method, the detection limits of dipyridamole and benzydamine were 345 pM and 147 nM in plasma, respectively. Diamino- and aminopyrenes were sensitively determined using TCPO and their detection limits were in the sub-fmol range [59], Carcinogenic compounds such as 1- nitropyrene and its metabolites, can also be determined by the HPLC-PO-CL system. Nonfluorescent nitropyrenes were converted into the corresponding fluorescent aminopyrenes by online reduction on a Zn column followed by detection 2-50-fmol detection limits were achieved in the determination of ethanol extracts from airborne particulates (Fig. 13) [60],... 

EVANGELISTA, R.A., LIU, M.-S., CHEN, F.-T.A., Characterization of 9-aminopyrene-l,4,6-trisulfonate-derivatize sugars by capillary electrophoresis with laser-induced fluorescence detection, Anal. Chem., 1995,67,2239-2245. 

HPLC on a Cosmosil 5 Cis column, using a perchloric acid-acetonitrile eluent (pH 7.6), followed by CLD in the presence of hydrogen peroxide and bis(2,4,6-trichlorophenyl) oxalate (42), was applied to the determination of 1-aminopyrene (43a) and various diaminopyrenes (43b-d). Ascorbic acid was added to avoid oxidative degradation of the aminopyrenes in the presence of metals LOD in the sub-fmol range (SNR 3)147. A fast (less than 10 min) HPLC-ELCD method was proposed for determination of dopamine (19b) and its metabolites in microdialysates, using packed fused silica capillary columns LOD 0.05 Xg/L of dopamine in a 2 XL sample, RSD 3% (n = 10)148. 

Glycoprotein was digested using the peptide N-Glycosidase F (PNGase F) to release N-linked glycan, then labeled with 8-aminopyrene-l,3,6-trisulfonate (APTS). The detection used LIF with a argon-ion laser. 

Suzuki, H., Muller, O., Guttman, A., and Karger, B. L. (1997). Analysis of 1-aminopyrene 3,6,8-trisulfonate-derivatized oligosaccharides by capillary electrophoresis with matrix- assisted laser desorptlon/ionization tlme-of-fhght mass spectrometry. Anal. Chem. 69, 4554—4559. 

Fig. 22 a and b. Circular dichroism and absorption spectra of 1-aminopyrene in mesophases 671 ... 

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