摘要
为揭示污水生物脱氮工艺中污泥菌群间的群体感应作用,建立了柱前衍生-固相萃取-高效液相色谱荧光检测法(HPLC-FLD)定量检测介导细菌种间群体感应信号分子AI-2的方法。取反应器的泥水混合液,经0.45μm滤膜过滤后,用氨基磺酸掩蔽亚硝酸盐干扰,并与2,3-二氨基萘(DAN)发生衍生化反应,衍生化产物用C18固相萃取柱进行固相萃取,经氮吹浓缩后上机分析。采用C18色谱柱(4.6 mm×250 mm,5μm)进行分离,乙腈与水(含0.1%甲酸)作为流动相进行梯度洗脱,使用荧光检测器(激发和发射波长分别为271 nm和503 nm)进行检测。结果表明,该检测方法在1~200 ng·mL~(-1)范围内呈现出了良好的线性关系,检出限为1 ng·mL~(-1),回收率为55.08%~59.25%,相对标准误差为2.98%~10.41%。该方法适用于杂质干扰多的痕量信号分子AI-2定量分析,可为生物脱氮工艺中信号分子AI-2介导群体感应研究提供有效的分析方法。
To reveal the mechanism of quorum sensing in biological nitrogen removal processes, a method usingpre-column derivatization, solid-phase extraction and high-performance liquid chromatography with fluorescencedetector(HPLC-FLD) was established for quantitation of AI-2 signal molecules. The samples collected from thereactor and filtered through 0.45 μm filter membrane, then were reacted with amino-sulfonic acid to mask nitriteand derived with 2,3-diaminonaphthalene(DAN). The derived products were extracted and concentrated by solidphase extraction(SPE), and then determined by HPLC(C18 column, 4.6 mm×250 mm, 5 μm) with fluorescencedetector(excitation wavelength and emission wavelength were 271 nm and 503 nm, respectively). Gradientelution was carried out by acetonitrile and water(containing 0.1% formic acid) as mobile phases. The resultspresented an excellent linearity with the concentration ranging from 1 to 200 ng·mL~(-1), and the detection limitswere 1 ng · mL~(-1). The recovery ratio of sample ranged from 55.08% to 59.25%, with the relative standarddeviations of 2.98% to 10.41%, respectively. This method provided an effective analytical method forquantitation of trace signal molecule AI-2 in biological nitrogen removal processes.
引文
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