Vatinno Rosa, Aresta Antonella, Zambonin Carlo G, Palmisano Francesco
Dipartimento di Chimica, Università degli Studi di Bari, Via E. Orabona 4, I-70126 Bari, Italy.
J Pharm Biomed Anal. 2007 Aug 15;44(4):1014-8. doi: 10.1016/j.jpba.2007.04.008. Epub 2007 Apr 19.
A new method for the determination of ochratoxin A (OTA) in human urine samples has been developed using solid-phase microextraction (SPME) interfaced with liquid chromatography-fluorescence detection (LC-FD). This method is simpler and cheaper compared to the most widely adopted clean-up procedures for OTA extraction from urine (usually based on immunoaffinity columns). Briefly, urine samples, diluted 1:5 with phosphate buffer (10 mM, pH 3), were partitioned against chloroform and the aqueous phase extracted by a polydimethylsiloxane/divinylbenzene (PDMS/DVB) fiber. The fiber was then "statically" desorbed, through a SPME interface, into a LC system operating in isocratic conditions. The linear range investigated in urine was 0.01-1 ng/ml. Within-day R.S.D.% in urine spiked at 0.1 and 1 ng/ml were 3.9 and 1.9, respectively, whereas the between-days R.S.D.% were 5.5 and 3.0, respectively. The limits of detection (LOD) and quantitation (LOQ) calculated at a signal-to-noise ratio of 3 and 10 (noise calculated peak to peak on a blank chromatogram at the OTA retention time) were 0.01 and 0.05 ng/ml, respectively.
已开发出一种使用与液相色谱 - 荧光检测(LC - FD)联用的固相微萃取(SPME)来测定人尿液样本中赭曲霉毒素A(OTA)的新方法。与从尿液中提取OTA最广泛采用的净化程序(通常基于免疫亲和柱)相比,该方法更简单且成本更低。简而言之,尿液样本用磷酸盐缓冲液(10 mM,pH 3)按1:5稀释,与氯仿进行分配,水相用聚二甲基硅氧烷/二乙烯基苯(PDMS/DVB)纤维萃取。然后,纤维通过SPME接口“静态”解吸到在等度条件下运行的LC系统中。在尿液中研究的线性范围为0.01 - 1 ng/ml。在添加了0.1和1 ng/ml OTA的尿液中,日内相对标准偏差(R.S.D.%)分别为3.9和1.9,而日间R.S.D.%分别为5.5和3.0。在信号噪声比为3和10(噪声在OTA保留时间的空白色谱图上峰峰值计算)时计算的检测限(LOD)和定量限(LOQ)分别为0.01和0.05 ng/ml。