Hayakawa K, Nakamura A, Terai N, Kizu R, Ando K
Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Chem Pharm Bull (Tokyo). 1997 Nov;45(11):1820-2. doi: 10.1248/cpb.45.1820.
Diesel exhaust particulates were extracted with benzene-ethanol (3:1, v/v) and separated into five fractions by silica-gel column chromatography. Direct-acting mutagenic activity was assayed by the Ames test using the Salmonella typhimurium YG1024 strain. The total activity of five fractions was about four times greater than that of the crude extract, suggesting that the activities in the fractions were suppressed in the crude extract. Strong activity was observed in fraction 4 which was eluted with dichloromethane (61.5% of the total activity) and fraction 5 which was eluted with ethanol (35.3%). Nitropolycyclic aromatic hydrocarbons (NPAHs) were determined by high-performance liquid chromatography with chemiluminescence detection. They were found mainly in fraction 4, although one NPAH was in fraction 3 which was eluted with n-hexane-dichloromethane (3:1, v/v). Based on these results, 53.1% of the activity in fraction 4 was attributed to NPAHs. The contribution of 1-nitropyrene and 1,3-, 1,6- and 1,8-dinitropyrenes was great and that of the other NPAHs was small. The mutagenic compound in fraction 5 was not identified. Fractions 1 and 2, which were eluted with n-hexane, and fraction 3 suppressed the activity of fraction 4. Polycyclic aromatic hydrocarbons in fractions 2 and 3 were considered as possible suppressors of NPAHs.
柴油废气颗粒用苯 - 乙醇（3:1，v/v）萃取，通过硅胶柱色谱法分离成五个馏分。使用鼠伤寒沙门氏菌YG1024菌株，通过艾姆斯试验测定直接作用诱变活性。五个馏分的总活性约为粗提物的四倍，这表明粗提物中馏分的活性受到抑制。在以二氯甲烷洗脱的馏分4（占总活性的61.5%）和以乙醇洗脱的馏分5（占总活性的35.3%）中观察到强活性。通过高效液相色谱 - 化学发光检测法测定硝基多环芳烃（NPAHs）。尽管有一种NPAH存在于以正己烷 - 二氯甲烷（3:1，v/v）洗脱的馏分3中，但它们主要存在于馏分4中。基于这些结果，馏分4中53.1%的活性归因于NPAHs。1 - 硝基芘以及1,3 -、1,6 - 和1,8 - 二硝基芘的贡献很大，而其他NPAHs的贡献较小。馏分5中的诱变化合物未被鉴定。以正己烷洗脱的馏分1和2以及馏分3抑制了馏分4的活性。馏分2和3中的多环芳烃被认为可能是NPAHs的抑制剂。
