沼液中溶解游离氨基酸的测定--柱前衍生-反相高效液相色谱法  

Determination of dissolved free amino acids in biogas slurry: RP-HPLC with pre-column derivation

作　　者：李建华[1] 刘文静[1] 李宁[1] 

机构地区：[1]同济大学环境科学与工程学院,污染控制与资源化研究国家重点实验室,上海200092

出　　处：《中国环境科学》2016年第8期2355-2363,共9页China Environmental Science

基　　金：国家自然科学基金项目(51308402,51538008);“水体污染控制与治理”国家科技重大专项(2013ZX07315-001);上海市科学技术委员会基础研究专项(13DJ1400104)

摘　　要：针对沼液特点开发pH值调节（pH 10.2）旋蒸浓缩（去除氨氮和挥发性生物胺）联合3K Millipore超滤离心分离的预处理方法.并以邻苯二甲醛（OPA）和氯甲酸芴甲酯（FMOC-Cl）为柱前衍生化试剂,结合反向高效液相色谱和荧光检测分析,对沼液中溶解游离氨基酸（DFAA）进行了定性和定量考察.该方法保证了各氨基酸在一定浓度范围内呈现良好的线性关系（R2均大于0.99）,标准样品中所选用的24种氨基酸,除谷氨酸回收率为70.5%外,其它氨基酸回收率为89%~115%,相对标准偏差为1.1%~5.0%.用所建立的方法对样品中溶解游离氨基酸的测定结果为：原料即剩余污泥中的溶解游离氨基酸量为2.0mol/L,5%和20%含固率反应器的沼液中的溶解游离氨基酸量分别为0.04mol/L和1.94mol/L,且种类不同,初步表明了不同厌氧消化反应系统中氨基酸的利用和产生机制的差异.In this study, a pretreatment protocol was developed, and combined with phthalaldehyde （OPA） and 9-fluorenyl methyl chloroformate （FMOC-C1） pre-column derivatization, RP-HPLC method, and fluorescent detection for the quantitative and qualitative investigations of dissolved free amino acids （DFAA） in biogas slurry. Owing to the simultaneous removal of high-strength ammonium and biogenic amines by pH adjustment during rotary evaporation, the recovery of DFAA was greatly improved by the introduction of 3K Millipore ultrafiltration tubes. As a result, a good linear relationship （corresponding linear correlation coefficient （R2） greater than 0.99） was obtained with the spike recovery rate of 23 kinds of amino acid of 89% -115%, except for that of glutamic acid of 70.5%. And the relative standard deviation was 1.1% -5.0% for the recovery analysis. On this basis, the developed pre-treatment method was further carried out to different sludge samples. The DFAA was 2.0mol/L in the excess sludge, while the amount of DFAA in the biogas slurry with input TS of 5% and 20% were different in species and amounts with 0.04mol/L and 1.94mol/L, respectively. Thus, no correlation between the DFAA amount and the input had been observed, which proved that the generation and utilization mechanisms of amino acids varied in different anaerobic digesters.

关 键 词：超滤 旋转蒸发 反相高效液相色谱 溶解游离氨基酸 沼液 

分 类 号：X703[环境科学与工程—环境工程]