Recovery of standard 125I-peptides at the picogram and femtogram level by reversed-phase high-performance liquid chromatography.
Recovery of standard 125I-peptides at the picogram and femtogram level by reversed-phase high-performance liquid chromatography.
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通过反相高效液相色谱法回收皮克和飞克水平的标准 125I 肽。
DOI：
10.1016/s0021-9673(01)92782-7
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Giese,RW
中科院分区：
文献类型：
--
作者：
Emanuel,RL;Williams,GH;Giese,RW
标准的[125I]血管紧张素II的平均回收率为92-103%，当使用C8键合硅胶填料的反相高效液相色谱(RP-HPLC)分析数量从10pg到50fg的这种多肽时，无论使用的是“旧”柱(先前使用一年)还是以前未使用过的柱。在一种情况下，在乙腈-0.01M高氯酸钠(40：60)的通常流动相上，该多肽在柱上的保留率为68%；相同的流动相，除0.1M高氯酸钠外，在该柱上的保留期缩短了10倍，平均回收率为103%。[125I]Brdykinin(105%)和[125I]促黄体激素释放激素(107%)的回收率相似，以1-PG水平测试。虽然两种商品化的[125I]血管活性肠肽的回收率仅为75%和85%，但高效液相色谱图也显示了几个峰。除了实际样品分析中可能出现的复杂情况外，这项工作还表明，在至少某些多肽的超微量分析中，反相高效液相色谱法是一种可行的样品清理/表征技术。
Average recoveries of standard [125I]angiotensin II are 92–103% when amounts of this peptide ranging from 10 pg to 50 fg are analyzed by reversed-phase high-performance liquid chromatography (RP-HPLC) employing a C8-bonded silica packing, whether an “old” (one year of prior use) or previously unused column is employed. In one case this recovery is 68% on a column giving a ten-fold higher retention of this peptide with the usual mobile phase of acetonitrile—aqueous 0.01Msodium perchlorate (40:60); the same mobile phase except 0.1Msodium perchlorate shortens the retention ten-fold on this column and gives an average recovery of 103%. Similar recoveries are obtained for [125I]brdykinin (105%) and [125I]luteinizing hormone releasing hormone (107%), tested at the 1 -pg level. Although two commercial preparations of [125I]vasoactive intestinal peptide are recovered only to the extent of 75 and 85%, the HPLC chromatograms also display several peaks. Aside from the potential for complications in the analysis of real samples, this work establishes that RP-HPLC is a viable technique for sample clean-up/characterization in the ultratrace analysis of at least some peptides.