Department of Pharmacology, College of Pharmacy, the Third Military Medical University, Chongqing 400038, China.
J Pharm Biomed Anal. 2012 Nov;70:447-55. doi: 10.1016/j.jpba.2012.06.022. Epub 2012 Jun 23.
CpG oligodeoxynucleotide 107 (CpG ODN107) could be used as a novel radiosensitizer for glioma. Herein, a novel and sensitive reversed-phase HPLC coupled with electrospray triple quadrupole mass spectrometry (LC-MS/MS) following a one-step C18 solid-phase extraction (SPE) for biological matrix removal was developed and fully validated for the determination of CpG ODN107 and its metabolites such as 5'N-1, 3'N-1, 3'N-2, and 3'N-3 in mouse plasma. The analytes were separated on an Extend-C18 analytical column (150 mm × 2.1 mm, 3.5 μm) using an eluent of acetonitrile-0.05% aqueous NH(3) (20:80, v/v) and detected by electrospray ionization (ESI) mass spectrometry in the negative multiple reaction monitoring mode (MRM). The assay was specific, and it showed a good linearity with a determination coefficient (r(2)) that was greater than or equal to 0.998 for CpG ODN107 and its metabolites in the biological matrices. The precision, accuracy, and relative recovery values were found to be <15%, ±15%, and 95-105%, respectively. This method was successfully applied to measure the concentrations of CpG ODN107 and its metabolites in the plasma following the intravenous administration of 15.0 mg/kg of CpG ODN107 in mice; therefore, the method was suitable for preclinical pharmacokinetic studies on CpG ODN107 and its metabolites.
CpG 寡脱氧核苷酸 107（CpG ODN107）可用作新型脑胶质瘤放射增敏剂。本研究建立了一种新型灵敏的反相高效液相色谱-电喷雾串联三重四极杆质谱（LC-MS/MS）法，并采用一步 C18 固相萃取（SPE）法用于生物基质去除，用于测定小鼠血浆中的 CpG ODN107 及其代谢物，如 5'N-1、3'N-1、3'N-2 和 3'N-3。分析物在 Extend-C18 分析柱（150mm×2.1mm，3.5μm）上分离，采用乙腈-0.05%氨水溶液（20:80，v/v）作为洗脱液，通过电喷雾电离（ESI）质谱在负多重反应监测模式（MRM）下检测。该方法特异性好，CpG ODN107 及其代谢物在生物基质中的线性测定系数（r2）均大于或等于 0.998。精密度、准确度和相对回收率分别小于 15%、±15%和 95-105%。该方法成功地应用于测定小鼠静脉注射 15.0mg/kg CpG ODN107 后血浆中 CpG ODN107 及其代谢物的浓度；因此，该方法适用于 CpG ODN107 及其代谢物的临床前药代动力学研究。
