Yang Ping, Li Xiaona, Yang Wenling, He Lian, Yang Li, Zhang Xianhua
Department of Pharmacy, Peking University Third Hospital, Beijing, 100191, China.
Department of Nephrology, Peking University Third Hospital, Beijing, 100191, China.
Rapid Commun Mass Spectrom. 2021 Jan 15;35(1):e8965. doi: 10.1002/rcm.8965.
Trimethylamine-N-oxide (TMAO) is a potential indicator of cardiovascular disease and chronic kidney disorders. It is important to monitor the TMAO level in plasma or serum of hemodialysis patients. A simple liquid chromatography/differential ion mobility spectrometry tandem mass spectrometry (HPLC/DMS-MS/MS) method was established and validated for the determination of TMAO in the serum of hemodialysis patients.
Chromatographic separation was performed on a Waters Atlantis HILIC silica column (2.1 × 50 mm, 3 μm). The gradient mobile phase consisted of 10 mM ammonium formate buffer and acetonitrile with 0.1% formic acid in both solvents. The serum sample was precipitated with acidic acetonitrile prior to HPLC/DMS-MS/MS analysis and TMAO-d was used as the internal standard. Data acquisition was performed in positive ion mode with a DMS system before the electrospray ionization source. The selected reaction monitoring transitions were m/z 76.0 → 58.0 and m/z 85.2 → 66.1 for TMAO and the internal standard, respectively.
Excellent linearity was observed over the calibration range 0.05-20 μg/mL (r > 0.995). The method was validated for good specificity and sensitivity. The inter-run and intra-run precision and accuracy were less than 13.6% and 10.7%, respectively.
We established a novel and robust HPLC/DMS-MS/MS method for the quantification of TMAO in human serum samples. The validated assay was simple, rapid, sensitive and reliable. The developed method could be applied to the assay of serum samples from patients with kidney disease who are undergoing hemodialysis.
氧化三甲胺（TMAO）是心血管疾病和慢性肾脏疾病的一个潜在指标。监测血液透析患者血浆或血清中的TMAO水平很重要。建立并验证了一种简单的液相色谱/差分离子淌度谱串联质谱法（HPLC/DMS-MS/MS）用于测定血液透析患者血清中的TMAO。
色谱分离在Waters Atlantis HILIC硅胶柱（2.1×50 mm，3μm）上进行。梯度流动相由10 mM甲酸铵缓冲液和乙腈组成，两种溶剂中均含有0.1%甲酸。血清样品在进行HPLC/DMS-MS/MS分析前用酸性乙腈沉淀，TMAO-d用作内标。在电喷雾电离源之前，使用DMS系统以正离子模式进行数据采集。TMAO及其内标的选定反应监测跃迁分别为m/z 76.0→58.0和m/z 85.2→66.1。
在校准范围0.05 - 20μg/mL内观察到良好的线性（r＞0.995）。该方法验证了具有良好的特异性和灵敏度。批间和批内精密度及准确度分别小于13.6%和10.7%。
我们建立了一种新颖且稳健的HPLC/DMS-MS/MS方法用于定量测定人血清样品中的TMAO。经过验证的测定方法简单、快速、灵敏且可靠。所开发的方法可应用于对正在接受血液透析的肾病患者血清样品的检测。
