Quantitative detection of nine phthalate metabolites in human serum using reversed-phase high-performance liquid chromatography-electrospray ionization-tandem mass Spectrometry
Quantitative detection of nine phthalate metabolites in human serum using reversed-phase high-performance liquid chromatography-electrospray ionization-tandem mass Spectrometry
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DOI：
10.1093/jat/27.5.284
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发表时间：
2003-07-01
影响因子：
2.5
通讯作者：
Barr, DB
中科院分区：
文献类型：
--
作者：
Kato, K;Silva, MJ;Barr, DB
我们开发了一种高灵敏度的方法来定量检测人血清中九种邻苯二甲酸酯代谢物。该方法需要在血液采集后立即使血清酶变性，以避免血液采集和储存过程中引入的污染物二酯母体化合物发生水解。在分析之前，对样品进行酶解结合以水解葡萄糖醛酸化的邻苯二甲酸酯单酯，并进行固相萃取以将单酯与其他血清成分分离。使用反相高效液相色谱-电喷雾电离-串联质谱法对提取物进行分析。血清中所有九种邻苯二甲酸酯单酯的检测限均在低纳克每毫升范围内 (0.6–1.3 ng/mL)。所有分析物的稳定同位素标记内标用于提高精度和回收率校正。这种高度选择性的方法可以分析邻苯二甲酸酯单酯，而不会因血清酶水解普遍存在的污染物邻苯二甲酸酯二酯而产生干扰。此外，它还可以直接测量据报道导致某些邻苯二甲酸酯的生殖和发育毒性的活性邻苯二甲酸酯单酯代谢物。
We developed a highly sensitive method for the quantitative detection of nine phthalate ester metabolites in human serum. This method requires denaturation of the serum enzymes immediately after blood collection to avoid the hydrolysis of the contaminant diester parent compounds introduced during blood collection and storage. Before analysis, the samples were subjected to an enzymatic deconjugation to hydrolyze the glucuronidated phthalate monoesters and a solid-phase extraction to isolate the monoesters from other serum components. The extracts were analyzed using reversed-phase high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry. The limits of detection of all nine phthalate monoesters in serum were in the low nanogram-per-milliliter range (0.6–1.3 ng/mL). Stable isotope-labeled internal standards for all analytes were used to improve precision and for recovery corrections. This highly selective method permits the analysis of phthalate monoesters without interferences resulting from the hydrolysis of the ubiquitous contaminant phthalate diesters by serum enzymes. In addition, it allows the direct measurement of the active phthalate monoester metabolites reportedly responsible for the reproductive and developmental toxicity of certain phthalates.