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采用高效液相色谱法(HPLC)对天目铁木不同时期的雄花序3种内源多胺—腐胺(Put)、精胺(Spm)、亚精胺(Spd)进行测定分析。结果发现:(1)适宜的苯甲酰化反应条件为37℃、30 min。(2)将色谱条件设定为检测波长254 nm,流动相配比甲醇︰水为60:40,流速0.6 mL.min-1,柱温30℃,进样量5μL。在该条件下内源多胺出峰和分离效果最佳。(3)在采样期间,雄花序中Spd含量最高,Spm次之,Put最少,且3种内源多胺含量均呈先升高后降低趋势,变幅明显。采用优化的HPLC法可有效快速地测定天目铁木雄花序的内源腐胺、精胺和亚精胺,并初步揭示了雄花序发育过程中3种内源多胺的变化趋势。
The contents of three endogenous polyamines, Put, Spm and Spd, of male inflorescences in different stages of Tianyu were determined by high performance liquid chromatography (HPLC). The results showed that: (1) The suitable conditions of benzoylation were 37 ℃ and 30 min. (2) The chromatographic conditions were set as detection wavelength 254 nm, mobile phase matching methanol: water 60:40, flow rate 0.6 mL.min-1, column temperature 30 ℃, injection volume 5μL. Under this condition, the peak of endogenous polyamines and the separation effect are the best. (3) During the sampling period, the content of Spd in male inflorescences was the highest, Spm was the second, Put was the least, and the contents of three endogenous polyamines were increased first and then decreased, and the amplitude was obvious. The optimized HPLC method can be used to determine the endogenous putrescine, spermine and spermidine in the male inflorescence of Tiemu, and the preliminary reveal of the trend of the three endogenous polyamines in the male inflorescence development.