Institute of Environmental Health, National Taiwan University, 17 Hsu-Chou Rd., Taipei 10055, Taiwan.
Talanta. 2012 Jan 30;89:237-45. doi: 10.1016/j.talanta.2011.12.020. Epub 2011 Dec 14.
This study developed and validated a method of measuring the feminizing chemicals 4-tert-octylphenol, 4-nonylphenol, nonylphenol monoethoxycarboxylate (NP(1)EC), nonylphenol monoethoxylate (NP(1)EO), nonylphenol diethoxylate (NP(2)EO), estrone, 17β-estradiol, estriol, 17α-ethinyl estradiol and bisphenol A in river water, sediment, and tissue using ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) and isotope-dilution techniques. Water samples were pretreated using disk-type automated solid-phase extraction (SPE). Solid samples of sediment, fish, and clams were treated with matrix solid-phase dispersion (MSPD) using C(8) adsorbent. Eluents were directly passed following alumina cartridges for cleanup. The signal intensity of analytes on electrospray ionization (ESI) was compared with that of atmospheric pressure photoionization (APPI). The analytes were separated on a UHPLC C(18) column with aqueous 10-mM ammonium acetate for NPEOs and aqueous 10-mM N-methylmorpholine for the other compounds. On-line cleanup was evaluated using two-dimensional liquid chromatography (2-D LC). ESI could provide satisfactory response for all of the analytes. Though APPI did not offer suitable response for NP(1)EO, NP(2)EO and NP(1)EC, it provided better signal intensities for the steroid estrogens (1.0-2.4 times) and the phenols (3.2-4.4 times) than ESI. UHPLC shortened chromatographic time to less than 10 min. Disk-type automated SPE and MSPD dramatically increased the throughput of sample preparation. The extraction efficiency on surface water samples ranged from 10% to 91%. The extraction efficiency of MSPD on sediment, fish, and clams was 51-101%, 36-109%, and 30-111%, respectively. Acidic alumina cleanup was essential for the analysis of the tissue sample, and reduced matrix effects better than 2-D LC on-line cleanup. The limits of detection (LODs) in water ranged from 0.81 ng/L to 89.9 ng/L. The LODs in sediment and tissue ranged from tens of pg/g wet weight to only a few ng/g wet weight. This method proved to be accurate and reproducible, as both quantitative biases and relative deviations remained smaller than 20% at three spiked levels.
本研究开发并验证了一种使用超高效液相色谱/串联质谱（UHPLC/MS/MS）和同位素稀释技术测量水中、沉积物中和组织中雌性化学物质 4-叔辛基苯酚、4-壬基酚、壬基酚单乙氧基羧酸酯（NP(1)EC）、壬基酚单乙氧基醚（NP(1)EO）、壬基酚二乙氧基醚（NP(2)EO）、雌酮、17β-雌二醇、雌三醇、17α-乙炔基雌二醇和双酚 A 的方法。水样采用盘式自动固相萃取（SPE）进行预处理。沉积物、鱼类和贝类的固体样品采用 C(8)吸附剂进行基质固相分散（MSPD）处理。洗脱液直接通过氧化铝柱进行净化。电喷雾电离（ESI）的分析物信号强度与大气压光离子化（APPI）的信号强度进行比较。分析物在 UHPLC C(18)柱上用 10-mM 乙酸铵水溶液分离 NPEOs，用 10-mM N-甲基吗啉水溶液分离其他化合物。在线净化采用二维液相色谱（2-D LC）进行评估。ESI 可以为所有分析物提供满意的响应。尽管 APPI 对 NP(1)EO、NP(2)EO 和 NP(1)EC 没有提供合适的响应，但它为甾体雌激素（1.0-2.4 倍）和酚类（3.2-4.4 倍）提供了更好的信号强度。UHPLC 将色谱时间缩短至不到 10 分钟。盘式自动 SPE 和 MSPD 极大地提高了样品制备的通量。对地表水样品的萃取效率范围为 10%-91%。MSPD 对沉积物、鱼类和贝类的萃取效率分别为 51-101%、36-109%和 30-111%。酸性氧化铝净化对于组织样品的分析至关重要，并且比在线 2-D LC 净化更好地降低了基质效应。水中的检测限（LOD）范围为 0.81ng/L 至 89.9ng/L。沉积物和组织中的 LOD 范围为几十 pg/g 湿重至仅几个 ng/g 湿重。该方法被证明是准确和可重复的，因为在三个加标水平下，定量偏差和相对偏差均保持在 20%以内。
