A simple and efficient method was developed for the preparation of high-purity trehalulose from the waste syrup of isomaltulose production. The waste syrup was pre-treated with C18 solid-phase extraction, where 98% decolorization and 97% reducing sugar recovery were obtained, followed by hydrophilic interaction liquid chromatography separation on a cysteine-bonded zwitterionic column. Under optimized conditions, trehalulose was separated from isomaltulose isomer and prepared on a semi-preparative scale with >99% purity. The structure of the prepared trehalulose was subsequently confirmed by nuclear magnetic resonance, and three tautomers of trehalulose (α-D-glucosylpyranosyl-1,1-β-D-fructopyranose, α-D-glucosylpyranosyl-1,1-β-D-fructofuranose, and α-D-glucosylpyranosyl-1,1-α-D-fructofuranose) were detected and completely characterized by ¹³C NMR spectroscopy for the first time in this study. The tautomerization of α-D and β-D type transition was observed by hydrophilic interaction liquid chromatography on an AdvanceBio Glycan Mapping column, with smaller particle size (2.7 μm). Furthermore, the prepared trehalulose was applied as a standard for trehalulose quantification during the sucrose conversion by Klebsiella sp. LX3. The combination of solid-phase extraction and hydrophilic interaction liquid chromatography offers a new avenue for the preparation of sugar isomers from complex natural or fermentation products.

中文翻译：

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采用固相萃取和亲水相互作用色谱法从异麦芽酮糖生产废糖浆中简单高效地制备高纯度海藻糖

建立了一种从异麦芽酮糖生产废糖浆中制备高纯度海藻糖的简便高效的方法。废糖浆经C18固相萃取预处理，脱色率98%，还原糖回收率97%，然后在半胱氨酸键合的两性离子柱上进行亲水相互作用液相色谱分离。在优化的条件下，海藻糖从异麦芽酮糖异构体中分离出来，并以 >99% 的纯度以半制备规模制备。随后通过核磁共振证实了制备的海藻糖的结构，海藻糖的三个互变异构体（α - D-吡喃葡萄糖基-1,1- β -D-吡喃果糖、α - D-吡喃葡萄糖基-1,1- β-D-呋喃果糖和α -D-吡喃葡萄糖基-1,1- α -D-呋喃果糖）在本研究中首次被¹³ C NMR光谱检测并完全表征。α- D 和β- D 型转变的互变异构是通过亲水相互作用液相色谱在 AdvanceBio 糖谱分析柱上观察到的，粒径更小 (2.7 μm)。此外，制备的海藻糖被用作克雷伯氏菌转化蔗糖过程中海藻糖定量的标准sp. LX3. 固相萃取和亲水相互作用液相色谱的结合为从复杂的天然或发酵产物中制备糖异构体提供了新的途径。