The present paper reports a new method for the quantification of the Ni2+-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% RSDPLSR. Validation of the Ni2+-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.
Keywords: IMAC; Ni2+-capacity; method validation; near-infrared spectroscopy; partial least squares regression.