利用HPLC/UV的方法偵測民眾尿液中cotinine含量的研究

本研究建立HPLC/UV的方法對cotinine進行分析，並應用於人體尿液中cotinine含量測定。利用高效能液相層析管柱Merck RP-18 column (250-4 mm, 5 μm)，醋酸-醋酸鈉緩衝溶液：甲醇(70：30, v/v)之混合溶液為移動相，UV波長設定在260 nm。於人體尿液中標準品分析所得之檢量線範圍為5 ng/mL ~ 50 ng/mL，相關係數(R2)大於0.999。於同日間與異日間分析方法之精密度與準確度，其相對偏差(R.S.D.)及相對誤差(R.E.)分別小於3.57 %及2.19 %，回收率大於91.96 %。本法應用對象分為社區民眾及國小學童。屏東縣琉球鄉及高雄市三民區56位民眾尿液分析結果，在主動吸菸者cotinine平均含量為3181.0±1797.8 ng/mL (n=7)；在家中或/及工作場所有受到二手菸污染威脅之民眾cotinine平均含量為315.3±793.3 ng/mL (n=19)；在家中及工作場所沒有受到二手菸污染者其cotinine平均含量為32.5±62.5 ng/mL (n=28)。針對高雄市某國小四、五、六年級學童作二手菸污染研究，有效問卷為588份 (95.1 %)。有380位學童(64.6 %)表示有受到二手菸的污染威脅，其中有325位學童表示在家中有受到二手菸污染的威脅(85.5 %)，有223位學童表示在父母親工作場所有受到二手菸污染的威脅(58.7 %)。依據父母親抽菸的情況、家中抽菸的人數、及二手菸暴露環境分成三部份，分層隨機抽樣收集105位學童的尿液並分析其cotinine含量。在父母親抽菸情況部份，其cotinine平均含量(ng/mL)可分成四組，分別為父母沒有抽菸7.56?b5.32、只父親抽菸17.89?b21.08、只母親抽菸6.87?b2.72及父母親都有抽菸25.16?b33.29。家中抽菸人數部分，從家中沒有人抽菸至家中有三人抽菸分成四組，其cotinine平均含量分別為7.61?b5.01、13.50?b16.63、15.62?b22.91、24.77?b27.29 ng/mL。學童二手菸暴露環境部分亦分成四組，分別為在家中、父母親工作場所、補習班(安親班)皆無二手菸暴露，只在家中暴露，只在父母親的工作場所及補習班(安親班)暴露，以及在家中、父母親工作場所、補習班(安親班)皆有暴露，其cotinine平均含量分別為6.97?b4.36、7.97?b5.74、13.96?b19.85、28.98?b18.25 ng/mL。

關鍵字

古丁尼；二手菸

並列摘要

An HPLC with UV detection have established for analysis of cotinine in urine. Mobile phase was consisted of acetate buffer and methanol(70 : 30, v/v). The cotinine was separated by Merck RP-18 column (250-4 mm, 5 μm). The wavelength of UV detector was set at 260 nm. The regression equations were linear between 5.0 ~ 50.0 ng/mL (R2 > 0.999). The R.S.D. and R.E. of intraday and interday were less than 3.57 % and 2.19 %, respectively. The recoveries were all greater than 91.96 %. This method was applied for determination cotinine contents in urine of community residents and elementary school students. The mean results of cotinine contents of 56 residents in Liouciou Township and Sanmin District as following, 3181.0±1797.8 ng/mL for active smokers (n=7), 315.3±793.3 ng/mL for the exposure to second hand smoke (SHS) at home or/and workplace (n=19), 32.5±62.5 ng/mL for non-exposure to SHS at home and workplace (n=28). The exposure to SHS of students of fourth, fifth and sixth grades in an elementary school was studied. 588 effective questionnaires were collected. 380 students (64.6 %) agreed to have an exposure to SHS. Among them, 325 students (85.5 %) had an exposure to SHS at home, 223 students (58.7 %) had an exposure to SHS at parents' workplace. The students were divided into three parts according to smoking situation of parents, number of smokers at home, and environments of SHS. By stratified random sampling, the urines of 105 students were collected and their contents of cotinine were analyzed. In part of smoking situation of parents, the mean contents of cotinine (ng/mL) are separated into following four groups: parents all not smoked 7.56±5.32, father smoked only 17.89±21.08, mother smoked only 6.87±2.72, and parents all smoked 25.16±33.29. In part of number of smokers at home, four groups were separated into as following: zero, one, two and three, the mean contents of cotinine (ng/mL) of above four groups are 7.61±5.01, 13.50±16.63, 15.62±22.91, 24.77±27.29, respectively. In part of environments of SHS, it was also separated into four groups as: non-exposure to SHS at home and workplace of parents and remedial class, exposure to SHS at home, exposure to SHS at workplace of parents and remedial class, exposure to SHS at home and workplace of parents and remedial class, the mean contents of cotinine (ng/mL) of above four groups are 6.97±4.36, 13.96±19.85, 7.97±5.74, 28.98±18.25 ng/mL, respectively.