Manganese concentrations in whole blood, plasma and serum of adult warmblood horses from 3 locations in Germany

Abstract

This study aimed to establish reference intervals for Mn in whole blood, plasma and serum of healthy, adult warmblood horses with known dietary Mn intake and to compare 2 methods of analysis. Between May 2018 and November 2019 a single blood sample was taken from a total of 270 clinically healthy horses (age: 3-25 years) in 3 stud farms. In lithium-heparin (LH) whole blood, LH plasma and serum Mn concentrations were analyzed by means of atomic absorption spectrometry (AAS) and inductively coupled plasma mass spectrometry (ICP-MS). The reference intervals were calculated according to the recommendations of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) and the Clinical and Laboratory Standards Institute (CLSI) comprising the range between 2.5 and 97.5 percentile of the analyzed Mn concentrations. Concurrent to blood sampling feed samples were taken and analyzed in order to determine the Mn content of the corresponding feeding ration. The horses' whole-blood Mn levels were significantly (p < 0.0001) higher with a median Mn concentration of 12.4 μg/l (reference interval: 4.99-25.1 μg/l [AAS]; 5.99-25.3 μg/l [ICP-MS]) than in the corresponding serum (median: 1.65 μg/l, reference interval: 0.60-3.50 μg/l [AAS]; 1.11-2.96 μg/l [ICP-MS]) or LH plasma (median: 1.35 μg/l, reference interval: 0.22-2.68 μg/l [AAS]; 0.59-2.45 μg/l [ICP-MS]). In the comparison of methods, there were statistically significant differences in the Mn determinations for LH plasma and serum between AAS and ICP-MS, whereas they yielded comparable results for whole blood. Whole blood shows an average of 10-fold higher Mn concentrations than serum or LH plasma. When evaluating blood Mn concentrations, consideration must be granted to the sample material under analysis as well as to which method is employed since relevant differences were found between AAS and ICP-MS in serum and LH plasma.