Department of Pharmaceutical Biosciences, Uppsala University, Uppsala, Sweden.
J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Jul 1;900:11-7. doi: 10.1016/j.jchromb.2012.05.014. Epub 2012 May 18.
A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the quantification of the opioid peptide DAMGO in rat plasma, as well as DAMGO and the microdialysis recovery calibrator [(13)C(2),(15)N]-DAMGO in microdialysis samples, is described. The microdialysis samples consisted of 15 μL Ringer solution containing 0.5% bovine serum albumin. Pretreatment of the samples involved protein precipitation with acetonitrile followed by dilution with 0.01% formic acid. The lower limits of quantification were 0.52 ng/mL and 0.24 ng/mL for DAMGO and [(13)C(2),(15)N]-DAMGO respectively and the response was linear up to 5000 fold higher concentrations. The plasma samples (50 μL) were precipitated with acetonitrile containing the isotope labeled analog [(13)C(2),(15)N]-DAMGO as internal standard. The method was linear in the range of 11-110,000 ng/mL. The separations were conducted on a HyPurity C18 column, 50×4.6 mm, 3 μm particle size, with a mobile phase consisting of acetonitrile, water and formic acid to the proportions of 17.5:82.5:0.01. Low energy collision dissociation tandem mass spectrometric (CID-MS/MS) analysis was carried out in the positive ion mode using multiple reaction monitoring (MRM) of the following mass transitions: m/z 514.2→453.2 for DAMGO and m/z 517.2→456.2 for [(13)C(2),(15)N]-DAMGO. The intra-day precision and accuracy did not exceed 5.2% and 93-104% for both compounds and sample types described. The inter-day precision an accuracy were <6.8% and 95-105% respectively. The method described is simple, reproducible and suitable for the analysis of small sample volumes at low concentrations.
一种液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）法，用于定量大鼠血浆中的阿片肽 DAMGO，以及微透析样品中的 DAMGO 和微透析回收率校准剂 [(13)C(2),(15)N]-DAMGO。微透析样品由 15 μL 包含 0.5%牛血清白蛋白的林格溶液组成。样品预处理包括用乙腈沉淀蛋白质，然后用 0.01%甲酸稀释。DAMGO 和 [(13)C(2),(15)N]-DAMGO 的定量下限分别为 0.52ng/mL 和 0.24ng/mL，响应线性高达 5000 倍更高的浓度。血浆样品（50μL）用含有同位素标记类似物 [(13)C(2),(15)N]-DAMGO 的乙腈沉淀作为内标。该方法在 11-110,000ng/mL 范围内呈线性。分离在 HyPurity C18 柱上进行，柱长 50×4.6mm，粒径 3μm，流动相由乙腈、水和甲酸组成，比例为 17.5:82.5:0.01。采用正离子模式，对质量转移进行多反应监测（MRM），进行低能量碰撞解离串联质谱（CID-MS/MS）分析：m/z 514.2→453.2 用于 DAMGO 和 m/z 517.2→456.2 用于 [(13)C(2),(15)N]-DAMGO。两种化合物和描述的样品类型的日内精密度和准确度不超过 5.2%和 93-104%。日间精密度和准确度分别小于 6.8%和 95-105%。所述方法简单、重现性好，适用于低浓度小体积样品的分析。
