大头金蝇成虫蝶啶荧光分析的方法学研究

Methodological Research of Pteridine Fluorescence Analysis in Adult Chrysomyia megacephala

目的:建立大头金蝇(Chrysomyia megacephala,C.megacephala)成虫头壳蝶啶含量的分析方法。方法:取C.mega-cephala头壳于3 mL Tris-HCl缓冲液(pH 8.0)中匀浆,离心,应用荧光分光光度计分析上清液的荧光强度。结果:当蝶啶溶液浓度低于0.8个头壳/3 mL时,其荧光强度随溶液浓度的增长呈线性增加(r2=0.9907,P<0.01)。浓度较高时,蝶啶溶液对其荧光有着显著的浓度淬灭效应,其荧光强度随溶液浓度增加逐渐降低(P<0.01)。光照条件下,蝶啶溶液的荧光强度随放置时间的延长而显著增加(r2=0.9676,P<0.01);黑暗条件下,虽然蝶啶溶液的荧光强度也随时间的延长而显著增加(P<0.05),但其增长速率比光照条件下小得多。结论:在进行蝶啶荧光分析时,必须控制蝶啶溶液的终浓度,注意避光,并缩短从匀浆到荧光分析的时间间隔,以减少分析误差。

Objective： To develop a method for pteridine fluorescence analysis in adult Chrysomyia megacephala（ C. megacephala）. Methods： Head capsules of adult C. megacephala were homogenized in 3 mL Tris-HCl buffer（pH 8.0）, centrifuged and analyzed using spectrofluorometer. Results： Fluorescence of the solution increased linearly with increasing concentration at the concentration of below 0. 8 head capsules per 3 mL（ r^2 = 0. 9907, P 〈 0.01 ）, and was obviously quenched at the higher concentration with the fluorescence decreasing significantly（ P 〈 0.01 ）. Fluorescence of the pteridine solution increased significantly with time in light（ r^2 = 0.9676, P 〈 0.01 ）. By contrast, fluorescence of the solution increased significantly（ P 〈 0.05）in darkness with much slower rate than that in light. Conclusion： During the course of pteridine fluorescence analysis, the final solution concentration must be limited. In addition, pteridine fluorescence analysis should be carried out free of light and be completed as soon as possible to reduce analytical errors.