臺灣博碩士論文加值系統

本研究利用離子對逆相高效能液相層析法配合紫外光-可見光偵測器測定組織樣品中所含之錳、銅與鈷。實驗結果證明以PAR為錯合試劑時，可成功地在波長450nm處測定Mn2+、Cu2+及Co2+。本研究流動相最佳條件為:甲醇/水(42.5/57.5；v/v)，流速為1.2 ml/min醋酸緩衝溶液濃度為8.0×10-4 mol L-1，離子對試劑(TBA-Br)濃度為8.0×10-3
mol L-1，pH=8.0，而錯合試劑濃度為3.0×10-4 mol L-1。偵測結果錳與鈷的線性範圍為0.1-3ppm，銅的線性範圍為0.3-7ppm。錳、銅與鈷的校正曲線分別為y=8.6957×105x+1.5522×105、y=6.4262×105x +0.2797×105與y=19.536×105x-0.1204×105。錳離子偵測極限為18.85ppb，銅離子偵測極限為7.37ppb，鈷離子偵測極限為0.32ppb。
牛肝標準品、豬肝臟、牛肝臟、豬腎及豬腦須先搗碎並烘乾以達到均質的效果，再經高溫爐灰化(800℃，6hr)以去除基質干擾，並使用草酸鈉為還原劑，還原在灰化過程形成的高氧化態錳。牛肝標準品 SRM 1577b的定量結果錳與銅含量分別為10.1±0.21與152.8±1.37，與確認值10.5 ±1.70及160 ±8.0相近，而相對誤差百分率分別為3.8%及4.5%，錳、銅與鈷的回收率分別為91.88±1.12%、92.1±1.01 %與91.94±1.09 %。市售組織樣品定量結果，相對標準偏差皆小於5%，回收率在90~100%。
綜合以上結果得知，利用PAR錯合試劑配合離子對液相層析法/紫外光-可見光偵測法，可成功地應用在偵測肝臟、腎臟及腦組織樣品中錳、銅及鈷元素的定性與定量分析，並具有快速、簡單及靈敏度高等優點。

An ion-pair reversed-phase high-performance liquid chromatographic method with UV-visible spectrophotometric detection is proposed for the simultaneous determination of manganese, copper and cobalt in biological samples. 4-(2-pyridylazo)resorcinol (PAR) chelates of Mn2+, Cu2+ and Co2+ were successfully separated and accurately determined at 450 nm. PAR chelates were eluted within 10 min at a flow-rate of 1.2 ml min-1 with a methanol aqueous mobile phase , CH3OH–water (42.5:57.5, v/v), containing 8.0×10-4 mol L-1 acetate buffer (pH 8.0), 8.0×10-3 mol L –1 TBA-Br and 3.0×10-4 mol L -1PAR. Under the optimum conditions, the linear range was 0.1-3 ppm for Mn2+ and Co2+, and 0.3-7ppm for Cu2+ with correlation coefficients (r2) being greater than 0.999. Calibration curves for Mn2+, Cu2+ and Co2+ were y=8.6957×105x +1.5522×105、y=6.4262×105x+0.2797×105 and y=19.536×105x -0.1204×105, respectively. The detection limits (S/N=3) for the chelates of Mn2+, Cu2+ and Co2+ were 18.85, 7.37 and 0.32 ppb, respectively.
For the purpose of homogeneity, the SRM 1577b, pig liver, bovine liver, pig kidney and pig brain were chopped to small pieces and baked till dryness. The samples were then ashed in elective furnace to remove the matrix interference. Sodium oxalate was used as reducing reagent to reduce the high oxidation state manganese from charred procedure. The proposed method provided a means for analyzing Mn2+ and Cu2+ in SRM bovine liver 1577b with experimental values of 10.1±0.21 and 152.8±1.37. The relative errors were 2.9% and 2.0%. The recoveries of Mn2+, Cu2+ and Co2+ were 91.88±1.12%, 92.1±1.01 % and 91.94±1.09 %, respectively. The RSD for tissue samples from market were lower than 5 %, and the recoveries were in 90~100 %.
In this research, the tissue samples were analyzed successfully after ashed pretreatment. The results were in good agreement with those obtained by flame atomic absorption spectrometry (FAAS).

目錄-----------------------------------------------------------------------I
中文摘要------------------------------------------------------------------IV
英文摘要------------------------------------------------------------------VI
圖目錄 -----------------------------------------------------------------VIII
表目錄---------------------------------------------------------------------X
第一章 緒論----------------------------------------------------------------1
第一節 錳、銅與鈷之簡介----------------------------------------------------3
一. 錳元素與其化合物之簡介-------------------------------------------------3
二. 銅元素與其化合物之簡介-------------------------------------------------4
三. 鈷元素與其化合物之簡介-------------------------------------------------5
第二節 微量元素之測定------------------------------------------------------7
一. 生物樣品---------------------------------------------------------------7
二. 動物組織樣品-----------------------------------------------------------9
第三節 高效能液相層析法---------------------------------------------------14
一. 原理------------------------------------------------------------------14
二. 離子對層析法(Ion-pair chromatography)---------------------------------15
三. 錯合試劑與應用--------------------------------------------------------17
第四節 研究動機與目的-----------------------------------------------------24
第二章 實驗部分-----------------------------------------------------------25
第一節 儀器設備-----------------------------------------------------------25
第二節 化學試劑與標準溶液-------------------------------------------------26
第三節 試劑溶液製備-------------------------------------------------------28
第四節 實驗步驟-----------------------------------------------------------28
第三章 分析方法之建立-----------------------------------------------------34
一. 錯合試劑的選擇--------------------------------------------------------34
二. 波長之選擇------------------------------------------------------------37
三. 緩衝溶液之pH值--------------------------------------------------------42
四. 金屬離子與PAR之錯合作用-----------------------------------------------50
五. PAR錯合試劑濃度與錯合時間---------------------------------------------52
六. 移動相之組成與流速----------------------------------------------------55
1. 離子對試劑的探討-------------------------------------------------------55
2. 有機溶劑的選擇與含量---------------------------------------------------58
3. 移動相流速的影響-------------------------------------------------------65
七. 干擾問題--------------------------------------------------------------66
第四章 真實樣品分析-------------------------------------------------------75
一. 樣品前處理------------------------------------------------------------75
二. 還原試劑之選擇--------------------------------------------------------79
三 結果與討論-------------------------------------------------------------85
(一). 肝臟組織------------------------------------------------------------85
1. 豬肝樣品的偵測---------------------------------------------------------85
2. 牛肝樣品的偵測--------------------------------------------------------100
(二). 豬腎臟組織---------------------------------------------------------104
(三). 豬腦組織-----------------------------------------------------------104
(四). 牛肝標準品的偵測---------------------------------------------------111
第五章 總結--------------------------------------------------------------115
參考文獻-----------------------------------------------------------------117

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