Yin Jing, Ni Kun-Yi, Shen Yue, Ma Peng-Cheng, Cao Ling, Wang Wei-Peng, Wang Yu
Department of Analytical Chemistry, China Pharmaceutical University, Nanjing 210009, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Sep 1;856(1-2):376-80. doi: 10.1016/j.jchromb.2007.06.033. Epub 2007 Jul 5.
A sensitive and specific method for determination of viaminate in human plasma by using high-performance liquid chromatography coupled with electrospray tandem mass spectrometry (LC-MS/MS) was developed in this study. The plasma samples were simply deproteinated, extracted, evaporated, and then reconstituted in 200 microl of methanol prior to analysis. Chromatographic separation was carried out on a Shimadzu VP-ODS column (250 mm x 2.0 mm, 5 microm) with a mobile phase of methanol-water (95:5, v/v) at a flow rate of 0.2 ml/min. Quantification was performed in the negative-ion electrospray ionization mode by selected ion monitoring of the product ions at m/z 164 for viaminate and m/z 109 for testosterone propionate which was used as the internal standard. The corresponding parent ions were m/z 446 and m/z 345. A linear calibration curve was observed within the concentration range of 0.10-200 ng/ml. The lowest limit of quantitation (LLOQ) was 0.1 ng/ml. The extraction-efficiency at three concentrations was 100.7, 93.6, and 99.7%. Practical utility of this new LC-MS/MS method was confirmed in pilot pharmacokinetic studies in humans following oral administration.
本研究建立了一种采用高效液相色谱-电喷雾串联质谱法（LC-MS/MS）测定人血浆中维胺酯的灵敏且特异的方法。血浆样品只需进行简单的去蛋白、萃取、蒸发，然后在分析前用200微升甲醇复溶。色谱分离在岛津VP-ODS柱（250 mm×2.0 mm，5微米）上进行，流动相为甲醇-水（95:5，v/v），流速为0.2毫升/分钟。在负离子电喷雾电离模式下，通过对维胺酯的m/z 164和用作内标的丙酸睾酮的m/z 109的产物离子进行选择离子监测来进行定量。相应的母离子分别为m/z 446和m/z 345。在0.10 - 200纳克/毫升的浓度范围内观察到线性校准曲线。最低定量限（LLOQ）为0.1纳克/毫升。三个浓度下的萃取效率分别为100.7%、93.6%和99.7%。在人体口服给药后的初步药代动力学研究中证实了这种新的LC-MS/MS方法的实际应用价值。
