Nguyen Phuong K, Owens Janel E, Lowe Luis E, Mooney Emily H
Department of Chemistry and Biochemistry, University of Colorado Colorado Springs, United States.
Department of Biology, University of Colorado Colorado Springs, United States.
MethodsX. 2020 Sep 2;7:101050. doi: 10.1016/j.mex.2020.101050. eCollection 2020.
Past analyses of sugar and amino acid composition of aphid honeydews have been completed using diverse instrumentation. Here we report the use of hydrophilic interaction liquid chromatography (HILIC) coupled with a triple quadrupole mass spectrometric (MS/MS) detector for the analysis of seven saccharides (xylose, fructose, glucose, sucrose, trehalose, melezitose and raffinose) and five amino acids (glutamic acid, glutamine, aspartic acid, serine, and asparagine). Limits of quantitation ranged from 0.05 mg/L (melezitose) to 1.0 mg/L (fructose) for sugars and from 0.10 mg/L (glutamic acid) to 3.66 mg/L (asparagine) for amino acids. Sample preparation was fast and simple, requiring only the washing of foils used to collect aphid honeydew with hot (80 °C) water and sonication of samples prior to HILIC/MS/MS analysis for both classes of analytes. No analyte derivatization was required and excellent chromatographic characteristics were observed. For those studying honeydew-mediated interactions in the field, this technique allows for rapid characterization of ecologically important amino acids and sugars.•Composition of seven saccharides in honeydew including xylose, fructose, glucose, sucrose, trehalose, melezitose,and raffinose, and five standard amino acids including glutamic acid, glutamine, aspartic acid, serine, and asparagine, were analyzed using hydrophilic interaction liquid chromatography-mass spectrometry.•All polar analytes were analyzed without derivatization using HILIC-MS with chromatographic run times of 7 min (sugars) and 10 min (amino acids).
过去对蚜虫蜜露中糖和氨基酸组成的分析是使用多种仪器完成的。在此，我们报告了使用亲水相互作用液相色谱（HILIC）与三重四极杆质谱（MS/MS）检测器联用，用于分析七种糖类（木糖、果糖、葡萄糖、蔗糖、海藻糖、松三糖和棉子糖）和五种氨基酸（谷氨酸、谷氨酰胺、天冬氨酸、丝氨酸和天冬酰胺）。糖类的定量限范围为0.05 mg/L（松三糖）至1.0 mg/L（果糖），氨基酸的定量限范围为0.10 mg/L（谷氨酸）至3.66 mg/L（天冬酰胺）。样品制备快速简单，对于两类分析物，在进行HILIC/MS/MS分析之前，只需要用热水（80°C）洗涤用于收集蚜虫蜜露的箔片，并对样品进行超声处理。无需对分析物进行衍生化，并且观察到了出色的色谱特性。对于那些在野外研究蜜露介导的相互作用的人来说，这项技术可以快速表征具有生态重要性的氨基酸和糖类。
• 使用亲水相互作用液相色谱 - 质谱法分析了蜜露中七种糖类（包括木糖、果糖、葡萄糖、蔗糖、海藻糖、松三糖和棉子糖）和五种标准氨基酸（包括谷氨酸、谷氨酰胺、天冬氨酸、丝氨酸和天冬酰胺）的组成。
• 所有极性分析物均未进行衍生化，使用HILIC - MS进行分析，色谱运行时间为7分钟（糖类）和10分钟（氨基酸）。
