Stir bar sorptive extraction with in situ de-conjugation and thermal desorption gas chromatography-mass spectrometry for measurement of 4-nonylphenol glucuronide in human urine sample
Stir bar sorptive extraction with in situ de-conjugation and thermal desorption gas chromatography-mass spectrometry for measurement of 4-nonylphenol glucuronide in human urine sample
复制标题
DOI：
10.1016/j.jpba.2005.05.024
复制
发表时间：
2006-01-23
影响因子：
3.4
通讯作者：
Nakazawa, H
中科院分区：
文献类型：
--
作者：
Kawaguchi, M;Ito, R;Nakazawa, H
采用搅拌棒吸附萃取（SBSE）和β -葡糖苷酶原位解偶联(TD)-气相色谱-质谱联用（GC-MS）技术对人尿样品中的4-壬基苯酚葡糖苷（NP-G）进行了分析。将蒸馏水（1 ml）、1.0 M醋酸铵溶液（100 μ l）和p -葡糖醛酸酶（10,000单位ml（-1）、10 μ l）加入人尿样品（1 ml）中，在37℃条件下提取90 min，在搅拌棒上涂有500 μ M厚的聚二甲基硅氧烷（PDMS）层，以250 rpm搅拌。然后，在选定的离子监测（SIM）模式下，对搅拌棒进行TD-GC-MS。以4-壬基酚（NP）为标准溶液，采用SBSE法绘制校准曲线。该方法在5 ~ 500 nM范围内线性良好，相关系数为0.999。此外，为了优化SBSE的原位解偶和回收率测试条件，本实验室采用生化技术合成了NP-G。NP的检出限（S/N= 3）和定量限（S/N bbb10）分别为0.2 ng ml(-1) （1.0 nM）和1.1 ng ml(-1) （5.0 nM）。在20 nM和100 nM的浓度下，NP-G在人尿样品（n = 6）中的平均加标回收率分别为104.1 （rsd = 7.1%）和100.6% (rsd = 9.2%)，并使用添加的内标4-（1-甲基）辛基酚d(5)进行校正。该方法能准确测定该标准，适用于人体尿液样品中微量NP-G的检测。(c) 2005 Elsevier B.V.版权所有
4-Nonylphenol glucuronide (NP-G) in human urine samples was analyzed using stir bar sorptive extraction (SBSE) with in situ deconjugation by beta-glucuronidase and thermal desorption (TD)-gas chromatography-mass spectrometry (GC-MS). Distilled water (1 ml), 1.0 M ammonium acetate solution (100 mu l) and P-glucuronidase (10,000 units ml(-1), 10 mu l) were added to human urine sample (I ml), and extraction was commenced for 90 min at 37 degrees C while stirring at 250 rpm with a stir bar coated with a 500-mu m-thick polydimethylsiloxane (PDMS) layer. Then, the stir bar was subjected to TD-GC-MS in the selected ion monitoring (SIM) mode. The calibration curve was made by SBSE method using 4-nonylphenol (NP) as the standard solution. The method showed good linearity and the correlation coefficients were 0.999 over the concentration range of 5-500 nM. Moreover, to optimize the conditions for SBSE with in situ de-conjugation and the recovery test, NP-G was synthesized by a biochemical technique in our laboratory. The limits of detection (S/N= 3) and quantitation (S/N > 10) for NP were 0.2 ng ml(-1) (1.0 nM) and 1.1 ng ml(-1) (5.0 nM), respectively. The average recoveries in the human urine samples (n = 6) spiked with NP-G at levels of 20 and 100 nM were 104.1 (R.S.D. 7.1%) and 100.6% (R.S.D. 9.2%), respectively, with correction using the added internal standard, 4-(1-methyl) octylphenol-d(5). The method enabled the precise determination of the standard and was applicable to the detection of trace amounts of NP-G in human urine samples. (c) 2005 Elsevier B.V. All rights reserved.