Quantitation of combinatorial libraries of small organic molecules by normal-phase HPLC with evaporative light-scattering detection
Quantitation of combinatorial libraries of small organic molecules by normal-phase HPLC with evaporative light-scattering detection
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DOI：
10.1007/bf01715529
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发表时间：
1996-08-01
影响因子：
3.8
通讯作者：
Kibbey, CE
中科院分区：
文献类型：
--
作者：
Kibbey, CE
蒸发光散射检测器的优点，紫外检测器的定量组合库组成的小有机化合物的高效液相色谱法进行了说明。该检测器的响应与样品发色团无关，这使得它非常适合于不同溶质混合物的色谱分析。因此，具有蒸发光散射检测的HPLC提供了在筛选测定中减少假阳性或假阴性结果的潜力，因为其能够检测在UV中吸收差的杂质（例如，源自聚合物载体的那些杂质）。此外，蒸发光散射检测器对类似结构类别的化合物表现出几乎等效的响应。因此，化合物文库的快速定量可以使用单一外标进行。例如，基于单个外标物，通过正相HPLC和蒸发光散射检测器对一系列类固醇、乙内酰脲以及BOC和Fmoc保护的氨基酸的定量误差平均约为+/-10%。蒸发光散射检测器的应用，定量低水平的样品杂质和检测器的兼容性与梯度洗脱也进行了说明。
The advantages of evaporative light-scattering detection over UV detection for the quantitation of combinatorial libraries composed of small organic compounds by HPLC are described. The detector's response is independent of the sample chromophore, which makes it well-suited to chromatographic analyses of mixtures of dissimilar solutes. Thus, HPLC with evaporative light-scattering detection offers the potential for reducing false positive or false negative results in screening assays, because of its ability to detect the presence of impurities that absorb poorly in the UV (e.g., those impurities originating from the polymeric support). Furthermore, the evaporative light-scattering detector exhibits a nearly equivalent response to compounds of similar structural class. Hence, rapid quantitation of compound libraries may be carried out with the use of a single external standard. For example, the quantitation errors, based on a single external standard, for a series of steroids, hydantoins, and BOC- and Fmoc-protected amino acids by normal-phase HPLC with evaporative light-scattering detection average approximately +/- 10%. The application of the evaporative light-scattering detector to the quantitation of low-level sample impurities and the detector's compatibility with gradient elution are also described.