US-SOMO HPLC-SAXS module: dealing with capillary fouling and extraction of pure component patterns from poorly resolved SEC-SAXS data.
US-SOMO HPLC-SAXS module: dealing with capillary fouling and extraction of pure component patterns from poorly resolved SEC-SAXS data.
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DOI：
10.1107/s1600576716011201
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发表时间：
2016-10-01
影响因子：
6.1
通讯作者：
Pérez J
中科院分区：
文献类型：
--
作者：
Brookes E;Vachette P;Rocco M;Pérez J
US-SOMO高效液相色谱小角度X射线散射模块是一种先进的综合分析SEC-SAXS(尺寸排除色谱联用SAXS)数据的工具。它包括基线和频带加宽校正例程，以及将重叠的偏斜峰的高斯分解为纯分量。尺寸排除层析与SAXS(小角X射线散射)联用，通常使用流动毛细管进行，应允许直接收集单分散样品数据。然而，毛细管结垢问题和非基线解析的峰可能会阻碍其有效性。超声扫描溶液模型器(US-SOMO)高效液相色谱-SAXS(高效液相色谱与SAXS联用)模块为分析此类数据提供了一个全面的框架，从简单的线性基线校正和对称的高斯分解工具开始[Brookes，Pérez，Cardinali，Proumo，Vachette&Rocco(2013)。J.Appl.克赖斯特。46,1823-1833]。除了几个新特点外，现已对这两个例行程序进行了实质性改进，包括利用先进的统计工具对结果进行评价。新的积分基线校正程序是基于一个更合理的假设，即毛细污垢对散射的影响随着X射线束内材料的散射强度单调增加。重叠的峰通常由于样品与列矩阵的相互作用而倾斜，现在可以使用非对称的修改的高斯函数来准确地分解。作为一个例子，分析了醛缩酶多分散溶液的情况：从高度曲折的峰中提取并可靠地模拟了四聚体、八聚体和十二聚体的SAXS谱。
The US-SOMO HPLC-SAXS (high-performance liquid chromatography coupled with small-angle X-ray scattering) module is an advanced tool for the comprehensive analysis of SEC-SAXS (size-exclusion chromatography coupled with SAXS) data. It includes baseline and band-broadening correction routines, and Gaussian decomposition of overlapping skewed peaks into pure components. Size-exclusion chromatography coupled with SAXS (small-angle X-ray scattering), often performed using a flow-through capillary, should allow direct collection of monodisperse sample data. However, capillary fouling issues and non-baseline-resolved peaks can hamper its efficacy. The UltraScan solution modeler (US-SOMO) HPLC-SAXS (high-performance liquid chromatography coupled with SAXS) module provides a comprehensive framework to analyze such data, starting with a simple linear baseline correction and symmetrical Gaussian decomposition tools [Brookes, Pérez, Cardinali, Profumo, Vachette & Rocco (2013). J. Appl. Cryst. 46, 1823–1833]. In addition to several new features, substantial improvements to both routines have now been implemented, comprising the evaluation of outcomes by advanced statistical tools. The novel integral baseline-correction procedure is based on the more sound assumption that the effect of capillary fouling on scattering increases monotonically with the intensity scattered by the material within the X-ray beam. Overlapping peaks, often skewed because of sample interaction with the column matrix, can now be accurately decomposed using non-symmetrical modified Gaussian functions. As an example, the case of a polydisperse solution of aldolase is analyzed: from heavily convoluted peaks, individual SAXS profiles of tetramers, octamers and dodecamers are extracted and reliably modeled.