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Effects of sample handling methods on substance P concentrations and immunoreactivity in bovine blood samples

Title: Effects of sample handling methods on substance P concentrations and immunoreactivity in bovine blood samples
Authors: Mosher, Ruby; Coetzee, Johann; Allen, Portia; Havel, James; Griffith, Gary; Wang, Chong
Contributors: Statistics
Source: archive/lib.dr.iastate.edu/stat_las_pubs/87/0-American_Veterinary_Medical_Association_permission.pdf|||Sat Jan 15 02:15:32 UTC 2022 ; archive/lib.dr.iastate.edu/stat_las_pubs/87/2014_Wang_EffectsSample.pdf|||Sat Jan 15 02:15:34 UTC 2022 ; 10.2460/ajvr.75.2.109
Publication Year: 2014
Collection: Digital Repository @ Iowa State University
Subject Terms: Large or Food Animal and Equine Medicine; Statistical Methodology; Veterinary Microbiology and Immunobiology; Veterinary Physiology
Description: Objective —To determine the effects of protease inhibitors and holding times and temperatures before processing on the stability of substance P in bovine blood samples. Samples —Blood samples obtained from a healthy 6-month-old calf. Procedures —Blood samples were dispensed into tubes containing exogenous substance P and 1 of 6 degradative enzyme inhibitor treatments: heparin, EDTA, EDTA with 1 of 2 concentrations of aprotinin, or EDTA with 1 of 2 concentrations of a commercially available protease inhibitor cocktail. Plasma was harvested immediately following collection or after 1, 3, 6, 12, or 24 hours of holding at ambient (20.3° to 25.4°C) or ice bath temperatures. Total substance P immunoreactivity was determined with an ELISA; concentrations of the substance P parent molecule, a metabolite composed of the 9 terminal amino acids, and a metabolite composed of the 5 terminal amino acids were determined with liquid chromatography–tandem mass spectrometry. Results —Regarding blood samples processed immediately, no significant differences in substance P concentrations or immunoreactivity were detected among enzyme inhibitor treatments. In blood samples processed at 1 hour of holding, substance P parent molecule concentration was significantly lower for ambient temperature versus ice bath temperature holding conditions; aprotinin was the most effective inhibitor of substance P degradation at the ice bath temperature. The ELISA substance P immunoreactivity was typically lower for blood samples with heparin versus samples with other inhibitors processed at 1 hour of holding in either temperature condition. Conclusions and Clinical Relevance —Results suggested that blood samples should be chilled and plasma harvested within 1 hour after collection to prevent substance P degradation. ; This article is from American Journal of Veterinary Research 75 (2014): 109, doi: 10.2460/ajvr.75.2.109 . Posted with permission.
Document Type: article in journal/newspaper
File Description: application/pdf
Language: English
Relation: archive/lib.dr.iastate.edu/stat_las_pubs/87/; 1084; 8849431; stat_las_pubs/87; https://dr.lib.iastate.edu/handle/20.500.12876/90689
Availability: https://dr.lib.iastate.edu/handle/20.500.12876/90689; https://hdl.handle.net/20.500.12876/90689
Accession Number: edsbas.DEC4BC07
Database: BASE