ANALYSIS OF OPIOID PEPTIDE ANALOGS

在该计划的前三年中积累的经验 项目赠款清楚地表明，需要在三个不同的领域进行改进， 该分析化学成分的区域。 A.每种肽的检测灵敏度将提高100倍， 1000倍，从皮摩尔到飞摩尔级别。近期初步 数据显示检测限为60 fmol。这种改进将使我们能够 以检测可能已经转移到 胎儿，并将允许感染少量的肽进入 母亲 B。经过司徒博士和克拉普筛选每个合成阿片肽类似物 为了确定阿片肽类似物是否具有适当的 生理活性，将收集血浆样品，并将肽 测定了HPLC纯化的肽将用质量 质谱法（MS）。以下讨论了关于MS定量的数据。 阿片肽类似物DALDA和DAMGO。相应的氘代 每种肽的合成类似物（由Schiller博士合成）将 用作MS定量的内标物，也用作载体 为了增加合成肽从生物样品中的回收， 矩阵这些分析数据将用于推导相应的 药代动力学参数 C.绵羊血浆样品的分析速度将提高10倍， 100-通过采用新的固相萃取（SPE）柱，绵羊 血浆将被直接应用，蛋白质沉淀将被 避免了

The experience gathered during the first three years of this program project grant clearly indicates the need for improvement in three separate areas for this analytical chemistry component. A. The detection sensitivity for each peptide will be improved 100-fold to 1000 fold, from the picomole to the femtomole level. Recent preliminary data show a limit of detection of 60 fmol. That improvement will enable us to detect any synthetic peptide that may have been transferred to the fetus, and will allow the infection of a smaller amount of peptide into the mother. B. After Drs. Szeto and Clapp screen each synthetic opioid peptide analog to determine whether that opioid peptide analog has appropriate physiologic activity, plasma samples will be collected and the peptide measured. The HPLC-purified peptide will be quantified with mass spectrometry (MS). Data are discussed below on the MS quantitation of the opioid peptide analogs DALDA and DAMGO. The corresponding deuterated synthetic analog of each peptide (synthesized by Dr. Schiller) will be used as the internal standard for MS quantification, and also as a carrier to increase the recovery of that synthetic peptide from the biological matrix. Those analytical data will be used to derive the corresponding pharmacokinetic parameters. C. The rate of analyzing ovine plasma samples will be improve 10-fold to 100-fold by employing new solid phase extraction (SPE) cartridges. Ovine plasma will be applied directly, and protein precipitation will be avoided.