Development and evaluation of an enzyme-linked immunosorbent assay kit for determination of benzo(a)pyrene in water
-
摘要: 目的 研制间接竞争酶联免疫吸附法(ELISA)检测苯并芘试剂盒。方法 采用棋盘滴定法确定抗原抗体的最佳用量;采用间接ELISA分析法确定苯并芘检测的最适条件,包括pH值、盐离子浓度及甲醇浓度。结果 抗原最佳包被浓度为10 μg/mL,单克隆抗体稀释倍数为 1:5000;在最优pH值为7.4、盐离子浓度为0.01 mol/L及甲醇浓度为10%的条件下,检测方法在5~50 ng/mL范围内线性相关,线性方程为y=-28.105 Ln(x)+123.66,R2=0.9937,IC50为13.74 ng/mL,苯并芘的最低检出限为3.3 ng/mL;在水样中的加标回收率为94.8%~112.3%;变异系数为4.38%~9.72%。结论 该方法适用于水体中苯并芘的检测,可用于大批量样品的快速筛查。
-
关键词:
- 苯并芘 /
- 酶联免疫吸附法(ELISA) /
- 抗原 /
- 抗体
Abstract: Objective To develop an indirect competitive enzyme-linked immunosorbent assay kit for the determination of benzo(a)pyrene(BaP).Methods The optimum concentration of antigen and monoclonal antibody were determined with chessboard titration.The optimum conditions,including pH,saltion and methanol concentration,were separately measured with indirect competitive enzyme-linked immunosorbent assay(icELISA).Results The optimum coating concentration of antigen was 10 μg/mL,while the optimum dilution ratio of antibody was 5 000.The established icELISA for the detection of BaP exhibited a linear range of 5 to 50 ng/mL,a sensitivity of 3.30 ng/mL,and a half inhibitory concentration(IC50)of 13.74 ng/mL in the conditions of pH 7.4,0.01 mol/L of saltion concentration and 10%of methanol.The average recovery was 105%in water samples.Conclusion These results indicate that the icELISA kit developed could be applied in rapid screening detection of BaP in water samples.-
Key words:
- benzo(a)pyrene /
- enzyme-linked immunosorbent assay /
- antigen /
- antibody
-
[1] 李玲.苯并芘对人支气管上皮细胞周期影响[J].中国公共卫生,2013,29(5):694-696. [2] 王丹,曹维强,王静.食品安全隐患-苯并(a)芘的研究进展[J].食品研究与开发,2006,27(1):132-135. [3] 刘新.饮用水中多环芳烃及其衍生物的分布和健康风险评价[D].重庆:西南大学硕士论文.2011. [4] Janoszka B.HPLC-fluorescence analysis of polycyclic aromatic hydrocarbons(PAHs)in pork meat and its gravy fried without additives and in the presence of onion and garlic[J].Food Chem,2011,126(3):1344-1353. [5] 杨红梅,王浩,刘艳琴,等.高效液相色谱法测定肉类食品中苯并芘残留的研究[J].食品研究与开发,2006,27(10):124-126. [6] 黄鸾玉,黎小正,秦振发.固相萃取/高效液相色谱荧光法测定水产品中苯并芘[J].分析实验室,2009,28(12)63-66. [7] 杨琳,陈青俊,欧菊芳,等.高效液相色谱法快速测定植物油中苯并[a]芘含量[J].粮食与油脂,2012,7(4):34-36. [8] Van der Wielen JC,Jansen JT,Martena MJ,et al.Determination of the level of benzo[a]pyrene in fatty foods and food supplements[J].Food Addit Contam,2006,23(7):709-714. [9] Frenich AG,Ocaa RM,Vidal JL.Determination of polycyclic aromatic hydrocarbons in airborne particulate matter by gas chromatography-triple quadrupole tandem mass spectrometry[J].J AOAC Int,2010,93(1):284-294. [10] Purcaro G,Moret S,Conte LS.Rapid validated method for the analysis of benzo[a]pyrene in vegetable oils by using solid-phase microextraction-gas chromatography-mass spectrometry[J].J Chromatogr A,2007,1176(1-2):231-235. [11] Hollosi L,Wenzl T.Development and optimisation of a dopant assisted liquid chromatographic-atmospheric pressure photo ionisation-tandem mass spectrometric method for the determination of 15+1 EU priority PAHs in edible oils[J].J Chromatogr A,2011,1218(1):23-31. [12] 王吕,邵辉峰,陈超超,等.苯并芘人工抗原的制备与鉴定[J].食品科学,2014,35(19):53-56. [13] 陈雪岚,金晶,杨晓慧.恩诺沙星快速酶联免疫吸附法检测试剂盒的研制[J].食品科学,2010,31(8):16-19.
点击查看大图
计量
- 文章访问数: 913
- HTML全文浏览量: 198
- PDF下载量: 164
- 被引次数: 0