Research Institute of Pharmaceutical Sciences, Musashino University, 1-1-20 Shinmachi, Nishitokyo, Tokyo 202-8585, Japan.
Anal Biochem. 2010 Jun 1;401(1):22-9. doi: 10.1016/j.ab.2010.02.022. Epub 2010 Feb 21.
A method for the quantification of acetylpolyamines, N(1),N(12)-diacetylspermine (DiAcSpm), monoacetylspermidine (AcSpd), and N(1),N(8)-diacetylspermidine (DiAcSpd), identifying each compound simultaneously, was developed with the goal of evaluating these acetylpolyamines as potential biomarkers of cancer. The method consists of prepurification of acetylpolyamines in urine with commercially available cartridges and derivatization with heptafluorobutyric (HFB) anhydride. HFB derivatives of acetylpolyamines were determined simultaneously using (15)N-labeled acetylpolyamines as internal standards by electrospray ionization and time-of-flight mass spectrometry (ESI-TOF MS). After the method was validated, the urinary acetylpolyamines of 38 cancer patients were quantified with this method. A comparison of the concentrations of DiAcSpm with those measured by a colloidal gold aggregation method demonstrated a correlation coefficient of 0.996, showing that the two methods were equally satisfactory. Analysis of the correlation between DiAcSpd or AcSpd and DiAcSpm, performed for the first time, indicated the usefulness of DiAcSpm as a urinary biomarker of cancer. During the course of this work, two simple methods for the preparation of alpha,omega-diacetylpolyamines were developed, and a possibility to separate and determine the concentrations of the two isomers, N(1)-acetylspermidine and N(8)-acetylspermidine in AcSpd, was shown by tandem mass spectrometry (MS/MS).
一种用于定量分析乙酰多胺，包括 N(1)，N(12)-二乙酰精胺 (DiAcSpm)、单乙酰腐胺 (AcSpd) 和 N(1)，N(8)-二乙酰腐胺 (DiAcSpd) 的方法被开发出来，目的是评估这些乙酰多胺作为癌症潜在生物标志物的可能性。该方法包括使用市售的试剂盒对尿液中的乙酰多胺进行预纯化，并用七氟丁酰 (HFB) 酐衍生化。通过电喷雾电离和飞行时间质谱 (ESI-TOF MS) 同时使用 (15)N 标记的乙酰多胺作为内标来确定 HFB 衍生物的乙酰多胺。在方法验证后，用该方法对 38 名癌症患者的尿液乙酰多胺进行了定量分析。DiAcSpm 浓度与胶体金聚集法测量结果的比较显示相关系数为 0.996，表明两种方法同样令人满意。首次对 DiAcSpd 或 AcSpd 与 DiAcSpm 之间的相关性进行分析，表明 DiAcSpm 作为癌症尿液生物标志物的有用性。在这项工作的过程中，开发了两种制备α，ω-二乙酰多胺的简单方法，并通过串联质谱 (MS/MS) 显示了分离和确定 AcSpd 中两种异构体，N(1)-乙酰腐胺和 N(8)-乙酰腐胺浓度的可能性。
