The amino acid glutamine (Gln) is a likely source of energy in the brain during neuroglucopenia. Effects of glucose deficiency on astrocyte Gln homeostasis remain unclear, as analytical tools of requisite sensitivity for quantification of intracellular levels of this molecule are not currently available. Here, a primary hypothalamic astrocyte culture model was used in conjunction with design of experiments (DOE)‐refined high‐performance liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI–MS) methodology to investigate the hypothesis that glucoprivation alters astrocyte Gln content in a sex‐specific manner. Critical mass spectrometric parameters for Gln derivative chromatographic response were identified by comparing the performance of central composite design, Box–Behnken design, and Optimal Design (OD)‐A, ‐D, ‐I, ‐Distance, and ‐Modified Distance DOE models. The outcomes showed that the OD‐A‐generated response was superior relative to other design outcomes. Forecasted surface plot critical mass spectrometric parameters were maximized by OD‐A, OD‐Distance, and OD‐Modified Distance designs. OD‐A produced a high‐performance method that yielded experimental run and forecasted surface plot maximal responses. Optimized mass spectrometric analysis of male versus female astrocyte Gln content provides novel evidence that glucoprivation significantly depletes this amino acid in female, but not in male, and that this sex‐specific response may involve differential sensitivity to estrogen receptor signaling. This technological advance will facilitate efforts to ascertain how distinctive physiological and pathophysiological stimuli impact astrocyte Gln metabolism in each sex.

中文翻译：

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通过星形胶质细胞谷氨酰胺测量的高性能实验设计优化 HPLC-电喷雾电离-质谱

氨基酸谷氨酰胺 (Gln) 可能是神经性葡萄糖减少症期间大脑中的能量来源。葡萄糖缺乏对星形胶质细胞 Gln 稳态的影响仍不清楚，因为目前还没有对该分子的细胞内水平进行量化所需的灵敏度的分析工具。在这里，将原代下丘脑星形胶质细胞培养模型与实验设计 (DOE) 结合使用——改进的高效液相色谱-电喷雾电离-质谱 (LC-ESI-MS) 方法来研究葡萄糖剥夺改变星形胶质细胞 Gln 含量的假设以特定性别的方式。通过比较中心复合设计、Box-Behnken 设计和最优设计 (OD)-A、-D、-I、-Distance、- 修正距离 DOE 模型。结果表明，OD-A 产生的响应优于其他设计结果。通过 OD-A、OD-Distance 和 OD-Modified Distance 设计使预测的表面图临界质谱参数最大化。OD-A 产生了一种高性能方法，该方法产生了实验运行并预测了曲面图的最大响应。男性与女性星形胶质细胞 Gln 含量的优化质谱分析提供了新的证据，表明葡萄糖缺乏会显着消耗女性体内的这种氨基酸，但男性不会，而且这种性别特异性反应可能涉及对雌激素受体信号传导的不同敏感性。这一技术进步将有助于确定独特的生理和病理生理刺激如何影响每个性别的星形胶质细胞 Gln 代谢。