臺灣博碩士論文加值系統

一、衍生化液相層析對硫氰酸離子及氰酸離子之同時微量分析研究
本研究嘗試以衍生化高效能液相層析法, 對硫氰酸離子及氰酸離子建立簡
單且高感度之微量同時分析方法。本法為異相反應系統乃利用相轉移催化
劑benzalkonium chloride(BAC) 將水層中硫氰酸離子與氰酸離子帶至甲
苯層, 再與甲苯層中化學可移除性衍生試劑2-(2-naphthoxy)ethyl
2-[1-(4-benzyl)piperazyl]ethane-sulfonate (NOEBPES) 進行衍生反
應, 形成具螢光特性之衍生產物; 所得之衍生物利用正相層析管柱
LiChrospher diolcolumn , 以含0.3% 異丙醇之 正己烷為移動相,
以7,12-dimethylbenz[a]anthracene內部標準, 利用紫外光檢出器(UV
detector) 在波長225 nm下進行偵測。本法已成功地應用於人體血漿中微
量硫氰酸離子及氰酸離子之含量分析。 影響硫氰酸離子及氰酸離
子衍生反應之諸多因素, 如系統中水相之酸鹼、相間轉移催化劑之種類及
需要量、反應時間及溫度、衍生試劑需要量等, 皆詳加探討。應用本法於
人體血漿中硫氰酸離子及氰酸離子之同時分析, 乃利用市售Ultrafree-MC
過濾器(30000NMWL polysulfone PTTK membrane) 在高速離心(
Ultrafiltration) 下取得濾液進行衍生反應, 以建立血漿中硫氰酸離子
及氰酸離子之同時分析。本法可測定血漿中微量硫氰 酸離子及氰酸離子
之線性範圍分別是0.4~20 nmol/mL 的硫氰酸離子及2~8 mmol/mL之氰酸離
子, 而其對硫氰酸離子與氰酸離子之偵測極限分別為每毫升 50 pmol 及
100 nmol (S/N = 3, 注入體積 10 mL)。其萃取相對回收率皆大於 95% ,
說明本法具有良好之選擇性及定量可靠性, 可用於人體血漿中微量硫氰酸
離子及氰酸離子之含量測定。二.衍生化液相層析對人體血漿中抗癲癇藥
物Ethosuximide之微量分析研究 本研究嘗試以衍生化高效能液相
層析法, 對抗癲癇藥物ethosuximide建立簡單且高感度之微量分析方法。
本法乃利用化學可移除性2-(2-naphthoxy)ethyl 2-[1-(4-benzyl)
piperazyl] ethanesulfonate (NOEBPES) 為衍生試劑, 以甲苯
(toluene) 為反應溶媒,氧化鎂 (MgO) 為鹼性催化劑, 進行勻相衍生反
應, 形成具螢光特性之衍生產物; 所得之衍生物利用正相層析管柱
LiChrospher diol column , 以含1.2% 異丙醇之正己烷溶液為移動相,
以coumarin 為內部標準, 利用紫外光檢出器在波長225 nm下進行偵測。
本法已成功地應用於人體血漿中抗癲癇藥物ethosuximide 之含量分析。
影響ESM衍生化反應之諸多因素, 如反應溶媒之篩選、鹼催化劑之種類及
需要量、反應時間及溫度、衍生試劑需要量等, 皆詳加探討。本法可應用
於人體血漿中ethosuximide之分析。本法可測定血漿中ESM 線性範圍
是30-700 nmol/mL, 而其偵測極限為每毫升 3 nmol (S/N = 3, 注入體積
10 mL)。其萃取相對回收率大於 94% , 說明本法具有良好之選擇性及定
量可靠性, 可用於人體血漿中ESM之含量測定。

1.Simultaneous determination of thiocyanate and cyanide anions
by derivatization and high performance liquid chromatography.
A sensitive high performance liquid chromatographic (HPLC)
method has been established for simultaneous determination of
thiocyanate and cyanide anions as their fluorogenic derivatives.
The method is based on the transfer of the anions from the
aqueous solution by benzalkonium chloride into the toluene
organic phase for derivatization with chemically removable
reagent, 2-(2- naphthoxy)ethyl 2-[1-(4-benzyl)
piperazyl]ethanesulfonate (NOEBPES). The derivatives obtained
were separated on a LiChrospher diol column with 0.3 %
isopropanol in n-hexane as the mobile phase and
7,12-dimethylbenz[a]anthracene as the internal standard. This
method has been successfully applied to the assay of
thiocyanate and cyanide anions in human plasma. Several
parameters affecting the partition/derivatization of thiocyanate
and cyanide anions such as base, the amount of phase transfer
catalyst, the reaction temperature and reaction time, the amount
of derivatizating agent were evaluated. Application of the
method to the analysis of thiocyanate and cyanide anions spiked
in human plasma, after simple ultrafiltration treatment, was
also studied. The linear range for the quantitation of
thiocyanate and cyanide anions in human plasma were over 0.4 ~
20 nmol/mL and 2 ~ 8 umol/mL, respectively. The detection
limits ( S/N = 3, sample size 10 mL) of thiocyanate and cyanide
anions were about 50 pmol/mL for thiocyanate anion and 100 nmol/
mL for cyanide anion, respectively. The intraday relative
standard deviation (n = 6) and the interday relative standard
deviation (n = 8) for thiocyanate anion were all less than
8.2%. The intraday relative standard deviation (n =!6) and the
interday relative standard deviation (n = 8) for cyanide anion
in plasma were all less than 2.9%. The relative recovery for
thiocyanate and cyanide anions in plasma were all greater than
95%.2.Determination of ethosuximide in plasma by derivatization
and high performance liquid chromatography A simple and
sensitive liquid chromatographic method is described for the
determination of ethosuximide in plasma, as a highly sensitive
derivative. The ethosuximide, spiked in plasma, after
separation with toluene extraction, was derivatized with
chemically removable derivatization reagent, 2-(2-naphthoxy)
ethyl 2-[1-(4-benzyl)piperazyl]ethanesulfonate (NOEBPES) in a
homogeneous system, using magnesium oxide as basic catalyst. The
resulting derivative was separated on a LiChrospher diol column
with 1.2 % isopropanol in n-hexane as the mobile phase and
coumarin as the internal standard. Several parameters
affecting the partition/derivatization of ethosuximide
extracted from spiked plasma such as the effect of organic
solvent, the amount of catalyst, the reaction temperature and
reaction time, the amount of derivatizating agent were
discussed.The linear range for the quantitation of ethosuximide
in spiked plasma was over 30 ~ 700 nmol/mL. The limit of
detection for ethosuximide in plasma was about 3 nmol/mL (S/N =
3, sample size 10 mL). The intraday relative standard
deviation (n = 6) and the interday relative standard deviation
(n = 6) were all less than 9.2% forumol/mL ethosuximide in
plasma. The relative recovery for ethosuximide in plasma was
greater than 94%. The method can be used for analysis of ESM in
human plasma for drug monitoring.