A new approach to study drugs containing ultra-high dilutions of antibodies using fluorescence spectroscopy

Published: 3 June 2020| Version 1 | DOI: 10.17632/2236svz2k8.1
Maksim Sapelnikov,
, Margarita Shishkina,


This data set contains results of ultrahigh dilutions effects study. The physicochemical (and therefore, spectral) properties of lactose water solutions change in the presence of ultrahigh dilutions of antibodies (UHD Ab). Spectrofluorimetric assessment of solutions containing UHD Ab to interferon gamma (IFNg)were performed and quantitative differences (420 nm peak emission intensity) from control samples were determined . The validation study of the procedure has shown that the proposed method is specific: UHD Ab IFNg samples differ significantly from samples that contain no UHD Ab IFNg (controls). A linear relationship between the measured parameter and the number of UHD Ab IFNg tablets used was also determined. Therefore, the method meets the validation criterion for linearity.The method has also been shown to provide precision and robustness. Therefore, according to the main criteria (specificity, detection limit, linearity precision and robustness) the method suggested for the identification and quantitative determination of UHD Ab IFNg presence in test samples is valid and can be applied for these purposes. Data processing is performed by means of the R language using the RStudio 1.2.5033 package. The initial fluorescence spectra are smoothed (interpolated) by splines. For this, a cubic spline with 42 nodes is used (the number of nodes is eight times less than the total number of points in the spectrum). The initial data are transformed according to a new basis, which leads to a smoothing of the initial curve. Local peaks are found as points of sign change of the second derivative of the transformed spectrum. Global peaks are selected among those local in proximity to the theoretical positions of the main peaks in the fluorescence spectrum. The data were analyzed statistically using one-way (assessment of specificity) or two-way (assessment of robustness) ANOVA, followed by Tukey’s post-hoc test for pairwise comparisons. The significance level (p) was set at 0.05.



Fluorescence Spectroscopy, Pharmaceutics, Lactose