Zhang Wenjing, Wan Sihui, Chen Lizhi, Wang Xuebin, Wang Zhuo, Huang Yi
Department of Pharmacy, Changhai Hospital of Shanghai, Second Military Medical University, Shanghai, People's Republic of China.
Department of Clinical Pharmacy, College of Pharmacy, China Pharmaceutical University, Nanjing, People's Republic of China.
Biomed Chromatogr. 2018 Jun;32(6):e4187. doi: 10.1002/bmc.4187. Epub 2018 Jan 29.
A new method for the analysis of cycloserine (4-amino-3-isoxazolidinone, CYC) in rat microdialysis samples has been developed. This method consists of derivatizing the CYC with benzoyl chloride, which transforms primary amines into highly stable derivatives. An attractive feature of this method was that the derivatization reaction is straightforward and can be completed within 10 min. The formed derivative, in contrast to the non-derivatized analyte, exhibited increased chromatographic retention and decreased matrix effects resulting from the co-elution of other components using reversed-phase liquid chromatography and on-line switching. Detection on a quadrupole-linear ion trap mass spectrometer (AB3200 Q-Trap) was performed using electrospray tandem mass spectrometry in multiple reaction monitoring mode. Various derivatization parameters were optimized in order to improve chromatographic separation and minimize ion suppression. In particular, the benzoylation reaction was improved to enhance the reproducibility and sensitivity of the chromatographic method. The transition m/z 207.1 → 105.1 was acquired to monitor the CYC derivatization products. The method was fully validated for its sensitivity, selectivity, matrix effect and stability. A good linearity over the selected range (r > 0.99, range = 22-2200 mg/L), as well as accuracy and precision within ±7% of the target values, was obtained. The assay described herein was successfully applied to quantitatively measure CYC in the lung and blood of anesthetized rats.
已开发出一种分析大鼠微透析样品中环丝氨酸（4-氨基-3-异恶唑烷酮，CYC）的新方法。该方法包括用苯甲酰氯将CYC衍生化，这会将伯胺转化为高度稳定的衍生物。此方法的一个吸引人的特点是衍生化反应简单直接，可在10分钟内完成。与未衍生化的分析物相比，形成的衍生物在反相液相色谱和在线切换条件下，由于其他成分的共洗脱，表现出更高的色谱保留率和更低的基质效应。在四极杆-线性离子阱质谱仪（AB3200 Q-Trap）上使用电喷雾串联质谱在多反应监测模式下进行检测。优化了各种衍生化参数，以改善色谱分离并最小化离子抑制。特别是，改进了苯甲酰化反应以提高色谱方法的重现性和灵敏度。获得了m/z 207.1 → 105.1的跃迁以监测CYC衍生化产物。该方法在灵敏度、选择性、基质效应和稳定性方面得到了充分验证。在选定范围内（r > 0.99，范围 = 22 - 2200 mg/L）具有良好的线性，以及在目标值的±7%以内的准确度和精密度。本文所述的测定方法已成功应用于定量测量麻醉大鼠肺和血液中的CYC。
