運用高效能液相層析法偵測人體尿液中古丁尼   全文下載

Determination of Cotinine in Human Urine by High-Performance Liquid Chromatography

一種逆相式液相層析法配合紫外光偵測來定量古丁尼的方法被發展出來。其移動相是由醋酸緩衝液、離子對試劑和甲醇來組成。古丁尼被一種碳18 形態的管柱將其與咖啡鹼、茶鹼、咖啡因、檳榔鹼和尼古丁於短暫的12 min 內進行良好的分離。紫外光偵測器的波長設在260 nm。對- 硝基苯胺被用來當作內部標準。人體尿液樣品可簡單的以液相- 液相萃取法來進行純化濃縮。標準樣品及生物樣品在濃度為0.1 - 10.0 μg/mL 間，兩者的迴歸方程式是呈直線關係（r > 0.999）。古丁尼的偵測極限為30.0 ng/mL。同日內與間日內之相對標準偏差和相對標準誤差分別小於3.93% 和3.07%（n = 6）。絕對與相對之回收率皆大於96.44%。本方法已成功的對一些主動吸菸者運用來偵測其尿液中的古丁尼。

A reversed-phase liquid chromatography with ultraviolet detection is described for the determination of cotinine. A mobile phase consisted of an acetate buffer with sodium octanesulfonate as an ion pair reagent and methanol was used to resolve cotinine from theobromine, theophylline, arecoline and nicotine. Using an octadecyl-type column, a good separation was achieved in 12 min. The wavelength of UV detector was set at 260 nm. With p-nitroaniline as internal standard, human urine samples were cleaned up by liquid-liquid extraction. The regression equations obtained from both the standard and the biological samples were linear between 0.1 - 10.0 μg/mL (r > 0.999). The detection limit for cotinine is 30.0 ng/mL. The R.S.D. and R.E. of intraday and interday are less than 3.93% and 3.07%, respectively (n = 6). The recoveries are all greater than 96.44%. This method has been successfully applied to the determination of urinary cotinine content in several heavy smokers.