Asp, Glu, and D-Ser are chiral amino acids and neurotransmitters binding to the N-methyl-D-aspartate receptor (NMDA) and they participate in glutamate signalization. D-amino acids are increasingly being recognized as important signaling molecules and variations in their levels are considered a marker of different pathologies, however, there is still a lack of knowledge about the role of most of D-amino acids in living organisms such as bone cells. A method for determination of concentrations of L/D-Asp, L/D-Glu and L/D-Ser in two types of bone cell lines: murine osteocytes (MLOY4) and osteoblasts (MC3T3-E1) is presented. It is based on capillary electrophoresis coupled to laser-induced fluorescence detection in normal polarity with 4-fluoro-7-nitro-2,1,3-benzoxadiazole as derivatizing agent suitable for an Argon ion laser source. The electrolyte consists of 137.5 mM borate buffer and 12.5 mM β-cyclodextrins as chiral selectors and the separation lasts 25 min. The method was optimized and validated for specificity, sensitivity, linearity, accuracy, and precision in murine osteocytes and osteoblasts. LLOQ was 0.25 µmol.L^-1 for the three D-amino acids and linearity was confirmed with r>0.995 for all D-and L-amino acids. Accuracy ranged between 81.9% to 111.7% and intra-day precision ranged between 1.8% to 10.9%. Concentrations of D- and L- Asp, Glu, and Ser are given and statistical differences between osteocytes and osteoblasts were found. The highest differences corresponded to L- and D-Glu. This method could play a fundamental role in the study of therapeutic targets in the treatment of bone diseases.

Asp，Glu和D-Ser是与N-甲基-D-天冬氨酸受体（NMDA）结合的手性氨基酸和神经递质，它们参与谷氨酸信号传导。D-氨基酸日益被认为是重要的信号分子，其水平的变化被认为是不同病理的标志，但是，仍然缺乏关于大多数D-氨基酸在生物（如骨骼）中的作用的知识细胞。提出了一种测定两种骨细胞系中的L / D-Asp，L / D-Glu和L / D-Ser浓度的方法：鼠类骨细胞（MLOY4）和成骨细胞（MC3T3-E1）。它基于毛细管电泳，耦合了正常极性下激光诱导的荧光检测，其中4-氟-7-硝基-2,1,3-苯并恶二唑为衍生剂，适用于氩离子激光源。电解质由137.5 mM硼酸盐缓冲液和12.5 mMβ-环糊精作为手性选择剂组成，分离过程持续25分钟。该方法已针对鼠类骨细胞和成骨细胞的特异性，灵敏度，线性，准确性和精密度进行了优化和验证。LLOQ为0.25 µmol.L三种D-氨基酸为^-1，所有D-和L-氨基酸的线性均r> 0.995。准确度在81.9％至111.7％之间，日内精确度在1.8％至10.9％之间。给出了D-和L-Asp，Glu和Ser的浓度，并发现了骨细胞和成骨细胞之间的统计学差异。最高差异对应于L-和D-Glu。该方法可能在研究骨疾病的治疗靶标中起重要作用。