Lee H G, Huffmon G V, Becklin R R, Tseng J L, Soble-Smith J, Robertson J T, Desiderio D M
Charles B. Stout Neuroscience Mass Spectrometry Laboratory, University of Tennessee at Memphis 38163, USA.
J Chromatogr B Biomed Appl. 1995 May 19;667(2):259-67. doi: 10.1016/0378-4347(95)00043-i.
Capillary reversed-phase high-performance liquid chromatography (RP-HPLC) was used to determine acetylmethylprednisolone (A-MP) that had been administered to feline spinal cord tissue. The method used a 300 mm x 0.32 mm I.D. packed capillary octadecylsilyl (ODS) column and an isocratic mobile phase of 40 mM triethylamine formate (TEAF, pH 3.2)-acetonitrile (50:50, v:v). The chromatographic behavior of A-MP was evaluated with respect to peak-area and peak-height by varying the A-MP concentration (12-190 microM) with a fixed injection volume (1 microliter), and by varying the injection volume (1-10 microliter) with a fixed concentration (12 microM) of A-MP. The limit of detection (signal-to-noise ratio, 3:1) was 250 pg (600 fmol) of synthetic A-MP. Various amounts of A-MP directly spiked into feline spinal cord segments were solvent extracted, separated, and plotted against peak-area (r2 = 1.00). Background tissue without A-MP gives minimal (< 1%) interference at 243 nm. The method also detects exogenous A-MP that was administered into feline spinal cord via an intrathecal injection. Furthermore, the presence of A-MP was confirmed via its molecular ion and corresponding product ions that were obtained by fast-atom bombardment tandem mass spectrometry (FAB-MS-MS).
采用毛细管反相高效液相色谱法（RP-HPLC）测定给予猫脊髓组织的乙酰甲基泼尼松龙（A-MP）。该方法使用内径为0.32 mm、长300 mm的填充毛细管十八烷基硅烷（ODS）柱和40 mM甲酸三乙胺（TEAF，pH 3.2）-乙腈（50:50，v:v）的等度流动相。通过在固定进样体积（1微升）下改变A-MP浓度（12 - 190 microM）以及在A-MP固定浓度（12 microM）下改变进样体积（1 - 10微升），从峰面积和峰高方面评估A-MP的色谱行为。检测限（信噪比为3:1）为合成A-MP的250 pg（600 fmol）。将直接添加到猫脊髓节段中的各种量的A-MP进行溶剂萃取、分离，并根据峰面积作图（r2 = 1.00）。不含A-MP的背景组织在243 nm处产生的干扰极小（< 1%）。该方法还能检测通过鞘内注射给予猫脊髓的外源性A-MP。此外，通过快速原子轰击串联质谱法（FAB-MS-MS）获得的分子离子和相应产物离子证实了A-MP的存在。
