建立一种采用柱前衍生化和 LC-MS/MS 检测人脑脊液中甘氨酸的方法。
Development of a method for the determination of glycine in human cerebrospinal fluid using pre-column derivatization and LC-MS/MS.
机构信息
Pharmacokinetics & Drug Metabolism, Amgen, Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
出版信息
J Pharm Biomed Anal. 2011 Sep 10;56(2):315-23. doi: 10.1016/j.jpba.2011.05.009. Epub 2011 May 18.
An LC-MS/MS method using pre-column derivatization with phenylisothiocyanate (PITC) was developed to quantify glycine in human cerebrospinal fluid (CSF) and applied to the determination of glycine in human samples collected during clinical testing. The calibration curve range for the assay was 50-10,000 ng/mL and ¹³C₂¹⁵N-glycine was used as an internal standard. Artificial CSF was used as a surrogate matrix for standards due to the presence of endogenous glycine in human CSF and this approach was validated with additional experiments involving either standard addition, or stable labeled glycine as an alternate calibration standard for endogenous glycine. Interday bias (% RE) and precision (% CV) were -4.2 and 12.3% at the LLOQ, and less than ±0.9 and 8.3% for higher concentrations, respectively. Glycine was stable in artificial CSF for at least 5h at room temperature, 55 days at -70 °C (-60 to -80 °C range), and through three freeze-thaw cycles.
建立了一种采用苯异硫氰酸酯（PITC）柱前衍生化的 LC-MS/MS 方法，用于定量人脑脊液（CSF）中的甘氨酸，并应用于临床测试中收集的人样品中甘氨酸的测定。该测定的校准曲线范围为 50-10,000ng/mL，¹³C₂¹⁵N-甘氨酸用作内标。由于人 CSF 中存在内源性甘氨酸，因此使用人工 CSF 作为标准的替代基质，并且通过涉及标准添加或稳定标记甘氨酸作为内源性甘氨酸替代校准标准的其他实验对该方法进行了验证。在LLOQ 时，日内偏差（%RE）和精密度（%CV）分别为-4.2%和 12.3%，而对于较高浓度，分别小于±0.9%和 8.3%。甘氨酸在人工 CSF 中至少在室温下稳定 5 小时，在-70°C（-60 至-80°C 范围）下稳定 55 天，并且通过三个冻融循环。
Zotero 插件