免疫亲和柱净化-高效液相色谱-串联质谱法测定母乳中的五氯苯酚

Determination of pentachlorophenol in breast milk by immunoaffinity column coupled with high performance liquid chromatography-tandem mass spectrometry

中文关键词:  高效液相色谱-串联质谱法  五氯苯酚  免疫亲和柱  母乳

英文关键词:high performance liquid chromatography-tandem mass spectrometry  pentachlorophenol  immunoaffinity column  human breast milk

中文摘要:

      目的 构建复合免疫亲和柱净化, 高效液相色谱-串联质谱法(high performance liquid chromatography- tandem mass spectrometry, HPLC-MS/MS)测定母乳中五氯苯酚残留的分析方法。方法 母乳样品用5%三乙胺乙腈水(7:3, V/V)提取, 经免疫亲和柱特异性净化, 以含0.05%氟化铵的甲醇和含0.05%氟化铵的超纯水为流动相, 梯度洗脱, ACQUITY UPLC HSS T3色谱柱分离, 多反应监测(multiple reaction monitoring, MRM)模式检测, 内标法定量分析。结果 五氯苯酚在0.02~20.00 μg/L范围内呈现良好线性关系, 相关系数达0.9998; 在低、中、高3个不同加标浓度下, 回收率在98.72%~106.95%之间, 相对标准偏差小于6.09%, 基质效应在94.45%~101.61%之间; 方法的检出限为0.008 μg/L, 定量限为0.02 μg/L。免疫亲和柱在该实验条件下至少可重复使用8次, 降低87%以上的实验成本。结论 本方法基于免疫亲和柱对样品进行特异性净化, 重现性好、准确度高, 能够有效地分离和检测母乳中的五氯苯酚残留, 可应用于母乳样品中五氯苯酚的定性定量检测。

英文摘要:

      Objective To construct an analytical method for the determination of pentachlorophenol residues in human breast milk by immunoaffinity column purification coupled with high performance liquid chromatography- tandem mass spectrometry (HPLC-MS/MS). Methods The human breast milk samples were extracted with 5% triethylamineacetonitrile-water (7:3, V/V), specifically cleaned by immunoaffinity column, with methanol containing 0.05% ammonium fluoride and ultrapure water containing 0.05% ammonium fluoride as mobile phases, gradient elution, separated by ACQUITY UPLC HSS T3 column, detected in multiple reaction monitoring (MRM) mode, and quantifed by internal standards. Results Pentachlorophenol showed a good linear relationship in the range of 0.02?20.00 μg/L, and the correlation coefficient reached 0.9998. The recoveries was 98.72%?106.95%, the relative standard deviations were less than 6.09%, and the matrix effects was 94.45%?101.61% under the 3 different spiked concentrations of low, medium and high. The limit of the detection was 0.008 μg/L and the limit of quantification was 0.02 μg/L. The immunoaffinity column could be reused at least 8 times under experimental conditions, reducing the experimental cost by more than 87%. Conclusion This method specifically cleans up samples based on the immunoaffinity column, which has good reproducibility and high accuracy, which can effectively isolate and detect pentachlorophenol residues in human breast milk, and can be applied to the qualitative and quantitative detection of pentachlorophenol in human breast milk samples.