摘要
在细胞色素C催化下,吡啰红B与青蒿素反应导致荧光降低,细胞色素C与青蒿素的反应为酶-底物模型。动力学研究表明,稳态催化速率依赖于酶和底物浓度,催化常数km、kmax及Kcat.分别为3.3×10^-5mol/L,5.4×10^-6mol·L^-1·s^-1和13.5s^-1,催化活性受去活化剂和乙醇抑制。在pH5.3、25℃及7.6×10^-7mol/L的细胞色素C催化条件下,荧光降低值△F(F0-F)与青蒿素浓度在7.1×10^-8~1.1×10^-6mol/L范围内呈线性关系;检出限为7.2×10^-9mol/L;加标回收率为96.3%~106.8%。方法已用于测定血浆和尿液介质中的微量青蒿素。
A method for the cytochrome C-catalyzed fluorescence determination of artemisinin was developed using pyronine B (PB) as substrate. The interaction between cytochrome C and artemisinin was an enzymesuhstrate model. The kinetic studies demonstrated that the steady-state catalytic rate depended upon enzyme and substrate concentrations, and the Michaelis-Menten parameters, Km, Vmax and Kcat were 3.3 × 10^-5 mol/L, 5.4 × 10^-6 mol · L^-1 · s^-1 and 13.5 s^-1, respectively. Catalytic activity of cytochrome C was inhibited in the presence of deactivated agents CN^- and ethanol. Under optimal conditions ( pH 5.3, 25℃ and 7.6 ×10^-7 mol/L of cytochrome C), fluorescence intensity change (F0 - F) of pyronine B was proportional to the artemisinin concentration from 7.1 × 10^-8 to 1.1 × 10^-6 mol/L, the detection limit (3σ) was 7.2 × 10^-9mol/L and recoveries are 96.3% - 106.8%. The proposed method was applied to detect the concentration of artemisinin in the media of plasma and urine.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2006年第2期173-177,共5页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金(No.20075012)
湖南省教委自然科学基金(No.02C313)资助项目
关键词
细胞色素C
青蒿素
吡啰红B
荧光降低值
Cytochrome C, artemisinin, pyronine B, fluorescence intensity change