Insects, due to their small size, have limited energy storage space, but they also have high metabolic rate, so their hemolymph sugars are incredibly dynamic and play a number of important physiological functional roles in maintaining energetic homeostasis. In contrast to vertebrates, trehalose is generally the primary sugar found in insect hemolymph, which is followed by glucose and fructose. Many analytical chemistry methods exist to measure sugars, yet a direct comparison of methods that can measure all three simultaneously, and trehalose in particular, from low sample volumes, are sparse. Using the honey bee as a model, we directly compare the leading current methods of using High Performance Liquid Chromatography (HPLC) with an evaporative light-scattering detector and Gas Chromatography coupled with Mass Spectrometry (GC–MS) to determine which method would be better for measuring trehalose, glucose, and fructose in terms of reproducibility, accuracy, and sensitivity. Furthermore, we injected the enzyme inhibitors trehalozin (a trehalase inhibitor) and sorbose (a trehalase p-synthase inhibitor) to manipulate the trehalose levels in honey bee foragers as a proof of concept that this sugar can be altered independently of hemolymph glucose and fructose levels. Overall the HPLC method was less reproducible for measuring fructose and glucose, and it also had lower sensitivity for measuring trehalose. Consequently, significant differences in trehalose levels within the forager class were only detected with the GC–MS and not the HPLC method. Lastly, using the GC–MS method in the follow up study we found that trehalozin and sorbose causes a significant increase and decrease of trehalose levels respectively, in forager honey bees, independent of the glucose and fructose levels, ten minutes after injection. Taken together, these methods will provide useful tools for future studies exploring the many different physiological functional roles that trehalose can play in maintaining insect energetic homeostasis.

昆虫由于体积小，能量存储空间有限，但是它们的代谢速率也很高，因此它们的血淋巴糖具有令人难以置信的动态性，并在维持能量稳态方面起着许多重要的生理功能。与脊椎动物相反，海藻糖通常是昆虫血淋巴中发现的主要糖，其次是葡萄糖和果糖。存在许多用于测量糖的分析化学方法，但是可以同时测量所有三个方法的方法的直接比较，尤其是海藻糖，尤其是从低样品量中获取时，是稀疏的。以蜜蜂为模型，我们直接比较了使用高效液相色谱（HPLC）和蒸发光散射检测器以及气相色谱与质谱（GC-MS）结合使用的领先方法，以确定哪种方法更适合测量海藻糖，葡萄糖和果糖在重现性，准确性和敏感性方面。此外，我们注入了酶抑制剂海藻糖蛋白（一种海藻糖酶抑制剂）和山梨糖（一种海藻糖酶对合酶抑制剂）来控制蜜蜂觅食者中的海藻糖水平，以证明这种糖可以独立于血淋巴葡萄糖和果糖水平而改变。 。总体而言，HPLC方法在测量果糖和葡萄糖方面的重现性较低，并且在测量海藻糖方面也具有较低的灵敏度。所以，在觅食类中，海藻糖水平的显着差异仅通过GC-MS而非HPLC方法检测到。最后，在后续研究中使用GC-MS方法，我们发现在注射后十分钟，海藻糖和山梨糖分别导致饲用蜜蜂的海藻糖水平显着增加和降低，而与葡萄糖和果糖水平无关。综上所述，这些方法将为今后的研究提供有用的工具，以探索海藻糖在维持昆虫体内能量稳态中所起的许多不同的生理功能作用。注射后十分钟，不依赖于葡萄糖和果糖水平的蜜蜂中的蜜蜂。综上所述，这些方法将为今后的研究提供有用的工具，以探索海藻糖在维持昆虫体内能量稳态中所起的许多不同的生理功能作用。注射后十分钟，不依赖于葡萄糖和果糖水平的蜜蜂中的蜜蜂。综上所述，这些方法将为今后的研究提供有用的工具，以探索海藻糖在维持昆虫体内能量稳态中所起的许多不同的生理功能作用。