Department of Biosystems, Katholieke Universiteit Leuven, Willem de Croylaan 42, Leuven, Belgium.
Phytochem Anal. 2010 Nov-Dec;21(6):602-8. doi: 10.1002/pca.1241.
S-adenosyl-l-methionine (SAM) plays an important role in many biochemical reactions in plants. It is mainly used as a methyl donor for methylation reactions, but it also participates in, for example, the biosynthesis of polyamines and the plant hormone ethylene.
To develop a fast capillary electrophoresis technique to separate SAM in fruits and fruit juices without any pre-purification steps.
Four different extraction solutions and two extraction times were tested, of which 5% trichloroacetic acid (TCA) for 10 min was found most suited. A glycine : phosphate buffer (200 : 50 mm, pH 2.5) was found optimal to analyse SAM in TCA extracts. Analyses were preformed on different climacteric and non-climacteric fruits and fruit juices. The calibration curve was created in degraded tomato extract. The CE-method was compared with a more conventional HPLC method described in literature.
The CE technique made it possible to completely separate the S,S- and R,S-diastereoisomeric forms of SAM. The CE method proved to be very fast (20 min total running time instead of 42 min) and more sensitive (limit of detection of 0.5 µm instead of 1 µm) compared with the conventional HPLC method.
Fast measurements of SAM in fruits and juices are favoured by capillary electrophoresis in a 200 : 50 mm glycine : phosphate (pH 2.5) buffer system.
S-腺苷甲硫氨酸（SAM）在植物的许多生化反应中起着重要作用。它主要用作甲基供体参与甲基化反应，但它也参与例如多胺和植物激素乙烯的生物合成。
开发一种快速毛细管电泳技术，无需任何预纯化步骤即可分离水果和果汁中的 SAM。
测试了四种不同的提取溶液和两种提取时间，其中发现 5%三氯乙酸（TCA）提取 10 分钟最适合。发现甘氨酸-磷酸盐缓冲液（200:50mm，pH2.5）最适合分析 TCA 提取物中的 SAM。在不同的呼吸跃变型和非呼吸跃变型水果和果汁中进行了分析。校准曲线是在降解的番茄提取物中创建的。将 CE 方法与文献中描述的更传统的 HPLC 方法进行了比较。
CE 技术能够完全分离 SAM 的 S，S-和 R，S-非对映异构体形式。与传统的 HPLC 方法相比，CE 方法具有更快的速度（总运行时间为 20 分钟，而不是 42 分钟）和更高的灵敏度（检测限为 0.5 µm，而不是 1 µm）。
在 200:50mm 甘氨酸-磷酸盐（pH2.5）缓冲液系统中，毛细管电泳有利于快速测量水果和果汁中的 SAM。
