高效液相色谱法测定茶鲜叶中游离多胺含量

Determination of free polyamines in fresh tea leaves by high performance liquid chromatography

建立高效液相色谱测定茶鲜叶中腐胺（Put）、亚精胺（Spd）和精胺（Spm）3种游离多胺的方法。样品经过67g/L聚乙烯聚吡咯烷酮（PVPP）吸附茶多酚，5％（体积分数）高氯酸低温提取，苯甲酰氯衍生后进行色谱分析，采用Wondasil C18（4．6mm×150mm）色谱柱，以甲醇和水为流动相进行梯度洗脱，流速为1．0mL／min。结果表明：3种多胺在5-60nmol／L浓度范围内具有良好的线性关系，相关系数r〉0．980，检出限为0．11～0．41nmol／L，定量限0．55-2．05nmol／L，加标回收率为88．5％-105．5％，相对标准偏差（RSD）为0．99％-7．20％。利用此方法分别对茶鲜叶中芽、叶和茎的多胺含量进行测定，发现3种样品中多胺均有检出。该方法准确、灵敏度高且重现性好，为茶鲜叶中多胺含量的研究提供检测手段。

We determined putrescine （ Put）, spermidine （Spd） and spermine （Spm） in fresh tea leaves by high performance liquid chromatography.The samples were extracted with 5% （V/V） perchloric acid （HC104） containing 67 g/L PVPP （w/V）, and then the three free-polyamines were derivatized with benzoyl chloride as derivatization agent. The polyamine derivatives were separated on a C 18 column by HPLC with ultrapure water and methanol as mobile phases at a flow rate of 1.0 mL/min. The linearity was excellent in the range between 5 and 60 nmol/L with the correlation coefficients above 0. 980. The limits of detection （ S/N= 3 ） were between 0. 11 and 0.41 nmoL/L, and the limits of quantification （ S/N= 10） were between 0.55 and 2. 05 nmoL/L. The recovery of standard addition was between 88.5% and 105.5% with the relative standard deviations （ n = 3） between 0. 99% and 7.20%. Based on this method, the 3 free polyamines were checked out in bud,leaf and stem of fresh tea leaves.The results indicated that the method is sensitive, efficient and accurate. It is suitable for the simultaneous determination of the 3 free polyamines in fresh tea leaves.