Institute of Analytical Chemistry and Radiochemistry, CCB-Center for Chemistry and Biomedicine, Innrain 80/82, 6020 Innsbruck, Austria.
ADSI-Austrian Drug Screening Institute, Innrain 66a, 6020 Innsbruck, Austria.
Molecules. 2018 Nov 24;23(12):3072. doi: 10.3390/molecules23123072.
The present paper reports a new method for the quantification of the Ni-capacity of an immobilized metal affinity chromatography (IMAC) material using near-infrared spectroscopy (NIRS). Conventional analyses using UV absorption spectroscopy or atomic absorption spectrometry (AAS) need to dissolve the silica-based metal chelate sorbent as sample pretreatment. In the first step, those methods were validated on the basis of an ideal homogenous NiSO₄-solution and unveiled that UV with an intermediate precision of 2.6% relative standard deviation (RSD) had an advantage over AAS with an intermediate precision of 6.5% RSD. Therefore, UV analysis was chosen as reference method for the newly established NIRS model which has the advantage of being able to measure the material directly in diffuse reflection mode. Partial least squares regression (PLSR) analysis was used as multivariate data analysis tool for quantification. The best PLSR result obtained was: coefficient of determination (R²) = 0.88, factor = 2, root mean square error of prediction (RMSEP) = 22 µmol/g (test-set validation) or 7.5% RSD. Validation of the Ni-capacity using UV absorption spectroscopy resulted in an intermediate precision of ±18 µmol/g or 5.0% RSD. Therefore, NIRS provides a fast alternative analysis method without the need of sample preparation.
本文报道了一种使用近红外光谱(NIRS)定量测定固定化金属亲和层析(IMAC)材料镍容量的新方法。传统的使用紫外吸收光谱或原子吸收光谱(AAS)的分析方法需要将基于二氧化硅的金属螯合吸附剂溶解作为样品预处理。在第一步中,这些方法基于理想的均相 NiSO₄溶液进行了验证,并揭示出中间精度为 2.6%相对标准偏差(RSD)的紫外光比中间精度为 6.5%RSD 的 AAS 具有优势。因此,紫外分析被选为新建立的 NIRS 模型的参考方法,该模型具有能够直接在漫反射模式下测量材料的优势。偏最小二乘回归(PLSR)分析被用作多元数据分析工具进行定量。获得的最佳 PLSR 结果为:决定系数(R²)=0.88,因子=2,预测的均方根误差(RMSEP)=22 µmol/g(测试集验证)或 7.5%RSD。使用紫外吸收光谱验证镍容量的中间精度为±18 µmol/g 或 5.0%RSD。因此,NIRS 提供了一种无需样品制备的快速替代分析方法。