Inoue K, Kato K, Yoshimura Y, Makino T, Nakazawa H
Hoshi University, Faculty of Pharmaceutical Sciences, Department of Analytical Chemistry, Tokyo, Japan.
J Chromatogr B Biomed Sci Appl. 2000 Nov 10;749(1):17-23. doi: 10.1016/s0378-4347(00)00351-0.
A simple and sensitive method using high-performance liquid chromatography with multi-electrode electrochemical detection (HPLC-ED) including a coulometric array of four electrochemical sensors has been developed for the determination of bisphenol A in water and human serum. For good separation and detection of bisphenol A, a CAPCELL PAK UG 120 C18 reversed-phase column and a mobile phase consisting of 0.3% phosphoric acid-acetonitrile (60:40) were used. The detection limit obtained by the HPLC-ED method was 0.01 ng/ml (0.5 pg), which was more than 3000-times higher than the detection limit obtained by the ultraviolet (UV) method, and more than 200-times higher than the detection limit obtained by the fluorescence (FL) method. Bisphenol A in water and serum samples was pretreated by solid-phase extraction (SPE) after removing possible contamination derived from a plastic SPE cartridges and water used for the pretreatment. A trace amount (ND approximately 0.013 ng/ml) of bisphenol A was detected from the parts of cartridges (filtration column, sorbent bed and frits) by extraction with methanol, and it was completely removed by washing with at least 15 ml of methanol in the operation process. The concentrations of bisphenol A in tap water and Milli-Q-purified water were found to be 0.01 and 0.02 ng/ml, respectively. For that reason, bisphenol A-free water was made to trap bisphenol A in water using an Empore disk. In every pretreatment, SPE methods using bisphenol A-free water and washing with 15 ml of methanol were done in water and serum samples. The yields obtained from the recovery tests using water to which 0.5 or 0.05 ng/ml of bisphenol A was added were 83.8 to 98.2%, and the RSDs were 3.4 to 6.1%, respectively. The yields obtained from the recovery tests by OASIS HLB using serum to which 1.0 ng/ml or 0.1 ng/ml of bisphenol A was added were 79.0% and 87.3%, and the RSDs were 5.1% and 13.5%, respectively. The limits of quantification in water and serum sample were 0.01 ng/ml and 0.05 ng/ml, respectively. The method was applied to the determination of bisphenol A in healthy human serum sample, and the obtained detection was 0.32 ng/ml. From these results, the HPLC-ED method should be the most useful in the determination of bisphenol A at low concentration levels in water and biological samples.
已开发出一种简单且灵敏的方法，即采用配备四个电化学传感器的库仑阵列的高效液相色谱 - 多电极电化学检测法（HPLC - ED）来测定水和人血清中的双酚A。为了实现双酚A的良好分离和检测，使用了CAPCELL PAK UG 120 C18反相柱以及由0.3%磷酸 - 乙腈（60:40）组成的流动相。HPLC - ED方法获得的检测限为0.01 ng/ml（0.5 pg），这比紫外（UV）法获得的检测限高3000多倍，比荧光（FL）法获得的检测限高200多倍。水和血清样品中的双酚A在去除源自塑料固相萃取小柱和预处理用水可能的污染后，通过固相萃取（SPE）进行预处理。通过用甲醇萃取，从小柱的各个部分（过滤柱、吸附剂床和筛板）检测到痕量（未检出至约0.013 ng/ml）的双酚A，并且在操作过程中用至少15 ml甲醇洗涤可将其完全去除。发现自来水中双酚A的浓度为0.01 ng/ml，超纯水（Milli - Q水）中为0.02 ng/ml。因此，使用Empore盘使不含双酚A的水来捕获水中的双酚A。在每次预处理中，水和血清样品均采用不含双酚A的水进行SPE方法，并用15 ml甲醇洗涤。向水中添加0.5或0.05 ng/ml双酚A后进行回收试验得到的回收率为83.8%至98.2%，相对标准偏差（RSD）分别为3.4%至6.1%。使用OASIS HLB对添加1.0 ng/ml或0.1 ng/ml双酚A的血清进行回收试验得到的回收率分别为79.0%和87.3%，RSD分别为5.1%和13.5%。水和血清样品中的定量限分别为0.01 ng/ml和0.05 ng/ml。该方法应用于健康人血清样品中双酚A的测定，获得的检测值为0.32 ng/ml。基于这些结果，HPLC - ED方法在测定水和生物样品中低浓度水平的双酚A时应是最有用的。
