Measuring haemolysis in cattle serum by direct UV-VIS and RGB digital image-based methods

Belén Larrán; Marta López-Alonso; Marta Miranda; Víctor Pereira; Lucas Rigueira; María Luisa Suárez; Carlos Herrero-Latorre

doi:10.1038/s41598-022-17842-4

Measuring haemolysis in cattle serum by direct UV-VIS and RGB digital image-based methods

Sci Rep. 2022 Aug 8;12(1):13523. doi: 10.1038/s41598-022-17842-4.

Authors

Belén Larrán^{1

2}, Marta López-Alonso³, Marta Miranda^{4

5}, Víctor Pereira³, Lucas Rigueira^{1

2}, María Luisa Suárez^{1

2}, Carlos Herrero-Latorre⁶

Affiliations

¹ Department of Anatomy, Animal Production and Clinical Veterinary Sciences, Faculty of Veterinary, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain.
² Veterinary Teaching Hospital "Rof-Codina", Faculty of Veterinary, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain.
³ Department of Animal Pathology, Faculty of Veterinary, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain.
⁴ Department of Anatomy, Animal Production and Clinical Veterinary Sciences, Faculty of Veterinary, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain. marta.miranda@usc.es.
⁵ Veterinary Teaching Hospital "Rof-Codina", Faculty of Veterinary, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain. marta.miranda@usc.es.
⁶ Research Institute on Chemical and Biological Analysis, Analytical Chemistry, Nutrition and Bromatology Department, Faculty of Sciences, University of Santiago de Compostela, Campus Terra, 27002, Lugo, Spain.

Abstract

A simple, rapid procedure is required for the routine detection and quantification of haemolysis, one of the main sources of unreliable results in serum analysis. In this study, we compared two different approaches for the rapid determination of haemolysis in cattle serum. The first consisted of estimating haemolysis via a simple direct ultraviolet-visible (UV-VIS) spectrophotometric measurement of serum samples. The second involved analysis of red, green, blue (RGB) colour data extracted from digital images of serum samples and relating the haemoglobin (Hb) content by means of both univariate (R, G, B and intensity separately) and multivariate calibrations (R, G, B and intensity jointly) using partial least squares regression and artificial neural networks. The direct UV-VIS analysis and RGB-multivariate analysis using neural network methods were both appropriate for evaluating haemolysis in serum cattle samples. The procedures displayed good accuracy (mean recoveries of 100.7 and 102.1%, respectively), adequate precision (with coefficients of variation from 0.21 to 2.68%), limit of detection (0.14 and 0.21 g L^-1, respectively), and linearity of up to 10 g L^-1.

MeSH terms

Animals
Calibration
Cattle
Hematologic Tests
Hemolysis*
Least-Squares Analysis
Neural Networks, Computer*