Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan, 430072, China.
Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Department of Chemistry, Wuhan University, Wuhan, 430072, China.
J Chromatogr A. 2019 May 10;1592:82-90. doi: 10.1016/j.chroma.2019.01.040. Epub 2019 Jan 14.
Trehalose-6-phosphate (T6P) is an important signaling metabolite that is involved in many physiological processes. However, the mechanism of the biological functions of T6P is not fully understood. Quantification of T6P in plants will be beneficial to elucidate the mechanism. However, it is still a challenge to chromatographically separate and sensitively detect T6P and related sugar phosphates. In the current study, we developed a method for effective separation and sensitive detection of glucose-1-phosphate (G1P), glucose-6-phosphate (G6P), sucrose-6-phosphate (S6P) and T6P in plant tissues by chemical derivatization combined with hydrophilic interaction liquid chromatography-tandem mass spectrometry (ChD-HILIC-MS/MS). With this method, two pairs of isomers (G1P/G6P and S6P/T6P) could be well separated on a HILIC column and sensitively detected by MS with limits of detection (LODs) ranging from 0.1 to 0.6 ng mL. The developed method was successfully applied to the detection of endogenous G1P, G6P, S6P and T6P in small amounts of plant tissues, such as 1 mg fresh weight of Oryza sativa shoot.
海藻糖-6-磷酸（T6P）是一种重要的信号代谢物，参与许多生理过程。然而，T6P 的生物学功能机制尚未完全阐明。对植物中的 T6P 进行定量分析将有助于阐明其机制。然而，色谱分离和敏感检测 T6P 及相关糖磷酸仍然是一个挑战。在本研究中，我们开发了一种通过化学衍生化结合亲水相互作用液相色谱-串联质谱（ChD-HILIC-MS/MS）有效分离和灵敏检测植物组织中葡萄糖-1-磷酸（G1P）、葡萄糖-6-磷酸（G6P）、蔗糖-6-磷酸（S6P）和 T6P 的方法。使用该方法，两对异构体（G1P/G6P 和 S6P/T6P）可以在 HILIC 柱上得到很好的分离，并通过 MS 进行灵敏检测，检测限（LOD）范围为 0.1 至 0.6ng/mL。该方法成功应用于检测少量植物组织（如 1mg 鲜重的水稻芽）中的内源性 G1P、G6P、S6P 和 T6P。
