昆虫体内多胺的高效液相色谱(HPLC)测定

Rapid analysis of polyamines in insects by high performance liquid chromatograpy.

建立丹磺酰氯柱前衍生HPLC快速测定昆虫体内多胺含量的方法。以C18(250mm×4.6mm,5μm)为固定相,甲醇和水为流动相,梯度洗脱,柱温40℃,流速1mL/min,荧光检测波长激发波长(Ex)280nm,发射波长(Em)515nm,测得腐胺(put)、亚精胺(spd)和精胺(spm)三者回收率分别为98.7%,99.2%和97.8%,回归方程线性良好(r值均大于0.99),分析时间为16min。该法简洁、快速、灵敏度高、重现性好,可有效分析昆虫及其他生物样品中微量多胺的含量。

A method for determinating polyamines in insects using high-poerformance liquid chromatographic（HPLC） was developed. This method was based on pre-column derivatization with dansyl chloride （Dns-Cl）. The derivatives were spearated on a C18 column（250 mm× 4.6 mm, 5 μm）, and eluted with methanol and distilled water using a onestep linear gradient.The temperature of folumn was 40℃ and the flow rate was 1 mL/min. The column eluate was monitored by fluorescence detection（excitatin： 280 nm ; emission ： 515 nm）. The recovery rate of three polyamines, putrescine（put）, spermindine（spd）, spermine（spin） were 98.7 %, 99.2 % and 97. 8 %, respectively. All the relative coefficients were above 0.99. The analysis can be finished in 16 min. Therefore, it is a simple, rapid and sensitive method that can be helpful in determining polyamines in insects and other biological tissues and body.