Namera Akira, So Angelina, Pawliszyn Janusz
Department of Chemistry, University of Waterloo, Ontario, Canada.
J Chromatogr A. 2002 Jul 19;963(1-2):295-302. doi: 10.1016/s0021-9673(02)00648-9.
A simple method for analysis of anatoxin-a in aqueous samples was developed using solid-phase microextraction (SPME) and high-performance liquid chromatography (HPLC) with fluorescence detection. Anatoxin-a was derivatized to a fluorogenic agent on the surface of the SPME fiber. In the method an SPME fiber was immersed for 30 min in the aqueous sample. The fluorogenic derivatizing reagent (4-fluoro-7-nitro-2,1,3-benzoxadiazole, 1.0 mg/ml in methanol) was dropped or sprayed onto the fiber containing extracted analytes. The fiber was then heated for 10 min in an empty vial at 70 degrees C in a waterbath to promote derivatization. The derivatives formed on the fiber were desorbed in a SPME-HPLC interface. The interface was filled with methanol-1 mM hydrochloric acid (7:3, v/v) before inserting of the fiber into the interface. For desorption, the fiber was inserted in the interface for 5 min. For anatoxin-a in an aqueous sample, the calibration curve showed linearity in the range of 50-1500 ng/ml and the limit of detection of anatoxin-a was 20 ng/ml. No interferences were found, and the time for analysis was 55 min for one sample.
开发了一种使用固相微萃取（SPME）和带荧光检测的高效液相色谱（HPLC）分析水样中天蓝蛋白-a的简单方法。天蓝蛋白-a在SPME纤维表面衍生化为荧光剂。在该方法中，将SPME纤维浸入水样中30分钟。将荧光衍生试剂（4-氟-7-硝基-2,1,3-苯并恶二唑，甲醇中浓度为1.0mg/ml）滴加或喷到含有萃取分析物的纤维上。然后将纤维在70℃的空瓶中于水浴中加热10分钟以促进衍生化。纤维上形成的衍生物在SPME-HPLC接口中解吸。在将纤维插入接口之前，接口中充满甲醇-1mM盐酸（7:3，v/v）。为了解吸，将纤维插入接口中5分钟。对于水样中的天蓝蛋白-a，校准曲线在50-1500ng/ml范围内呈线性，天蓝蛋白-a的检测限为20ng/ml。未发现干扰，一个样品的分析时间为55分钟。
